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目次

コンファレンスプログラム ·············································································· 2
発表演題及び講演時間 ······················································································ 3
シンポジウム講演要旨 ···················································································· 11
一般講演要旨···································································································· 40
ポスター発表講演要旨 ···················································································· 50
人名索引············································································································ 82
糸状菌分子生物学研究会会則········································································· 85
糸状菌分子生物学研究会運営委員名簿 ························································· 86
Ở

2

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第 5 回糸状菌分子生物学コンファレンスプログラム
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日時：平成 17 年 11 月 7 日（月）-９日（水）会場：東京大学弥生講堂（東京都文京区弥生１−１−１）
主催：糸状菌分子生物学研究会
後援：糸状菌遺伝子研究会
11 月 7 日（月）13:00-18:00
18:00-

シンポジウム「タンパク質工場としての麹菌の高度利用」
懇親会（農学部生協食堂）

11 月 8 日（火）9:30〜12:00
12:00〜13:30
13:30〜15:30
15:30〜17:15
17:15〜17:25

シンポジウム「糸状菌の有性生殖、無性生殖」
昼休み
ポスター発表（P-1〜P-34）
一般講演（O-1〜O-7）
総会

11 月 9 日（水）10:30〜12:00
12:00〜13:30
13:30〜15:30
15:30〜17:00
17:00〜17:05

一般講演（O-8〜O-13）
昼休み
ポスター発表（P-35〜P-64）
一般講演（O-14〜O-19）
閉会の辞

3発表演題および講演時間
ࢨࣤ࣎ࢩ࣑ࢗ 11 ᭮ 7 ᪝㸝᭮㸞13:00-18:00
ࠔࢰࣤࣂࢠ㈻ᕝሔ࡛ࡊ࡙ࡡ㯔Ⳟࡡ㧏ᗐฺ⏕ࠕ භത ⏍∸⣌≁ᏽ⏐ᴏᢇ⾙◂✪ᨥᥴࢬࣤࢰ࣭
13:00

ࠔࡢࡋࡴ࡞ࠕ໪ᮇ ົࡥࡆ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞

13:05

S-1ࠔ㯔ⳞⓆ⌟⏕ࣈࣚࢪ࣐ࢺࡡ㉰㎷㏷షᠺἪ࡛ ࡐࡿ ࢅ⏕࠷ࡒࣆࢡࢰࣤࣂࢠ㈻ࡡ⏍⏐ࠕ᭯ᒱ Ꮥ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞

13:25

S-2ࠔ㯔Ⳟ࡞ࡻࡾࣃࢹࣛࢯࢲ࣭࣑⏍⏐࡞ཀྵ࡯ࡌ ࣈࣞ ࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪࡡຝᯕࠕ୷ᒜ ₮ୌ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞

13:45

S-3ࠔ㯔Ⳟ⣵⬂ባࡡ࣓ࣛࢸࣛࣤࢡ㸯⣵⬂හ࡞⏍ ⏐ࡈ ࡎࡒࢰࣤࣂࢠ㈻ࢅᇰ㣬୕ΰ࡫ᅂ཭ࡌࡾ
ࡒࡴࡡᡋ␆ࠕᇷහ ⿩஄㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞

14:05

S-4ࠔ㯔Ⳟࡡᾦమᇰ㣬࡚ᅖమᇰ㣬࡛ྜྷᵕࡡ㐿ఎ ᏄⓆ ⌟ࢅᐁ⌟ࡌࡾࡒࡴࡡᡋ␆ࠕᆊᮇ ࿰ಆ㸝㒿㢦⥪ྙ◂✪ᡜ㸞

14:25

S-5ࠔࣈࣞࢷ࢛࣭࣑࠾ࡼぜࡒ㯔Ⳟࡡࢰࣤࣂࢠ㈻ ⏍⏐ ᶭᵋࠕᒷୖ ࿰⿩࣬⧂⏛ ೸㸝㒿㢦⥪ྙ◂✪ᡜ㸞

14:45

㸝ఆ ៹㸞

14:55

S-6ࠔࢤࢺࣤ᭩㐲໩࡞ࡻࡾ㯔Ⳟࡡ␏⛸ࢰࣤࣂࢠ ㈻⏍ ⏐ຝ⋙ࡡྡྷ୕ࠕ஫࿝ ົஒ㸝᮶໪ኬ㝌࣬㎨࣬⏍∸⏐ᴏ๭ᠺ⛁Ꮥ㸞

15:15

S-7ࠔAspergillus ᒌ⣊≟Ⳟࡡࢰࣤࣂࢠ㈻ኬ㔖⏍⏐ࢨ ࢪࢷ࣑㸯㧏㌹෕࣬㧏⩳ゼ⣌ࡡ㛜Ⓠࠕᖶ⏛ ᪺⏍㸝ኬ㛭࣬⥪◂㸞

15:35

S-8ࠔ㯔Ⳟࡡኣᙤ࡝∸㈻⏍⏐ࡡࡒࡴࡡ᩺ぞᐙ୹ ࣊ࢠ ࢰ࣭⣌ࡡ㛜Ⓠࠕ
▴⏛ ༡ᶖ㸝᭮᰿෗࣬⥪◂㸞

15:55

S-9ࠔHigh level production of biopharmaceuticals in the fungus Aspergillus oryzaeࠕ
Hjort, C.㸝Fungal Gene Technology, Novozymes A/S, Denmark㸞

16:30

㸝ఆ ៹㸞

16:40

S-10ࠔGenomic analysis of protein secretion and associated stresses in AspergillusࠕArcher, D. B.㸝University of Nottingham, UK㸞

17:35

ࠔࣂࢾࣜゞㄵࠕ

ㅦⁿ⩽ධဤ

17:55

ࠔ⤂ࢂࡽ࡞ࠕ⚽⏛ ಞ㸝㒿㢦⥪ྙ◂✪ᡜ㸞

4ࢨࣤ࣎ࢩ࣑ࢗ 11 ᭮ 8 ᪝㸝ℾ㸞9:30ࠤ12:00
ࠔ⣊≟Ⳟࡡ᭯ᛮ⏍Ṣ࣬↋ᛮ⏍Ṣࠕ
 Sexual and asexual development in filamentous fungi
9:30

S-11ࠔᏄᄙⳞ࠽ࡻࡦ୘ᏰධᏄᄙⳞࡡஹ㒼࡛ஹ㒼ᆵ 㐿ఎᏄࠕ᭯Ờ ງࠉ㔘Ꮔ ຉ 1 ࠉ㔘᲻ḿᶖ 2 ࠉຊ⸠ࣀࢻ 3ࠉ஬ᒜ┷⌾ 4ࠉᐣᶌຊዄࠉᑈᒱ ᚥ㸝᮶ா㎨ᕝኬᏕኬᏕ㝌࣬1 ⌟ ㎨ᴏ⎌ሾᢇ⾙◂✪ᡜ࣬2 ⌟ ࢦࣤࢹ࣭ࣛ඙㐅ᢇ⾙ᚺ⏕◂✪ᡜ࣬3 ⌟ TIS࣬
4

10:00

⌟ JICA㸞

S-12 Mating Type Genes-Mediated Sexual Reproduction in Filamentous Ascomycetes
Sung-Hwan Yun㸝Soonchyunhyang University, Korea㸞

10:30

S-13 Sexual Development, Pheromon Receptor in Aspergillus nidulans
Kap-Hoon Han㸝PaiChai University, Korea㸞

11:00

S-14 Mating Genes in Aspergillus oryzae Genome: Can It have a Sexual Life Cycle?

Nanase Yamamoto1, Praveen Rao Juvvadi1, Jun-ichi Maruyama1, David B Archer2, Katsuhiko
Kitamoto1 (1Department of Biotechnology, The University of Tokyo, 1-1-1, Yayoi, Bunkyo-Ku, Tokyo
113-8657, Japan. 2School of Biology, University of Nottingham, University Park, Nottingham, NG7
2RD, UK.)
11:30

S-15 ࠔAspergillus nidulans ࡞࠽ࡄࡾ୔ࡗࡡ mitogen-activated protein kinase (MAPK) ⤊㊪ࠕཿᕖ೸ኯ㑳ࠉ㜷㒂ᩏᝃ㸝᮶໪ኬ㝌㎨࣬ᚺ⏍⛁㸞5ୌ⯙ㅦⁿ㸝O-1㹳O-7㸞 11 ᭮ 8 ᪝㸝ℾ㸞15:30ࠤ17:15
15:30

O-1

㯔Ⳟࡡࣆ࢔ࣜࢰ࣭ᇰ㣬᮪௲ୖ࡚ࡡⓆ⌟よᯊఫ㔕ඔ᫓㸡ᑚᯐலᕵᏄࠉ⏣⏛㞖஄*ࠉኬ⟺ಘୌ 㸝㔘Ἁᕝኬࠉ*⏐⥪◂㸞

15:45

O-2

ࢷ࣒ࣞ࢓௛ຊ࣊ࢠࢰ࣭࡞ࡻࡾ㯔Ⳟᙟ㈻㌹ᥦ ࡛ᑙථ DNA ࡡよᯊᴃᮇ៿ୌࠉཿᕖ⫩௥ࠉᮄᮟኣỜࠉᮿୖ┷⏜⨶ࠉ㕝ᮄ⪵ࠉ᯵ᮄ㇇ࠉ㸝㣏⥪◂㸞

16:00

O-3

⣊≟Ⳟ࣬㓕ẍ࡞࠽ࡄࡾ RNA ࢦ࢕ࣝࣤࢨࣤࢡ⤊㊪ࡡ㐅໩Ⓩよᯊ୯ᒁᩔᆍ㸡みㆺ├ᶖ㸡Nguyen Bao Quoc㸡┶ᒜ⁘ᚷ㸝♼ᡖኬ࣬㎨Ꮥ㒂㸞

16:15

O-4

EGFP ࡛ AoVam3p ࡡ⼝ྙࢰࣤࣂࢠ㈻ࢅ⏕࠷ࡒᾦ⬂ ⭯ࡡྊっ໩࡞ࡻࡾ㯔Ⳟ
A. oryzae ᾦ⬂ࡡᙟឺよᯊḿ㊪ῗஒ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞

16:30

O-5

㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪ⤊ ㊪ࡡ よᯊᵵཾ⿩ḗ㑳ࠉ୯℀ᬓ஄ࠉḿ㊪ῗஒࠉ᭯ᒱ Ꮥࠉ໪ᮇົࡥࡆ (᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ)

16:45

O-6

㯔Ⳟ(Aspergillus oryzae)ࡡ impala ᵕ DNA ࢹࣚࣤࢪ࣎ ࢯࣤ Aoimp1 ࡡⳞᰬ㛣ኣᵕᛮ࡛㌹⛛Ὡᛮᑚ➗ཋ༡ಘ㸦㸡㸧㸡ᑚ⏷ ᾀ㸨㸡⛑ Ὂ஦㸨㸡㧏ᶣ▹⧂㸧㸡஫࿝ົஒ㸦㸝㸦᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ㸡㸧⚽⏛┬⥪㣏◂࣬⏍∸ᶭ⬗㸡㸨᭮᰿෗࣬⥪◂㸞

17:00

O-7

ࢠ࢙ࣤ㓗⏍⏐⣊≟Ⳟ Aspergillus niger ࡞࠽ࡄࡾὩᛮ 㓗⣪⛸࡞ᑊࡌࡾ⪇ᛮࡡ alternativeoxidase ࡡᐞ୙᭱㒂㈏⃀ࠉᮄ㔕㑝ჹࠉ᱊ᮟකኯ㑳 㸝᪡ኬ࣬⌦ᕝ࣬ᚺ໩㸞ୌ⯙ㅦⁿ㸝O-8㹳O-13㸞 11 ᭮ 9 ᪝㸝Ề㸞10:30ࠤ12:00
10:30

O-8

࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ APSES ࢰࣤࣂࢠ㈻ Mstu1 ࡢ௛╌ჹࡡᠺ⇅ࢅโᚒࡌࡾ
こᮟ㯖㔓Ờ㸡ᯐ㛏⏍ 㸝⏍∸◂㸞

10:45

O-9

Cryphonectria parasitica ࡡ GTP ⤎ྙࢰࣤࣂࢠ㈻ βγ-ࢦࣇࣗࢼࢴࢹ࡛ࣆ࢚ࢪࢸ࣭ࣖࢨࣤ➗ཋ ⣸ 㸝ᐋᇖኬ࣬㣏⏐ᴏ࣬⎌ሾࢨࢪࢷ࣑㸞

11:00

O-10

࣊ࢹ⑋Ⳟ, Phytophthora infestans, ࡞࠽ࡄࡾ఩Ὼ࡞ᚺ➽ࡊࡒ㐿ఎᏄⓆ⌟โᚒᶭᵋ࡞ࡗ࠷࡙
ㆺಞ἖*㸡 Howard S. Judelson㸝Dept. of Plant Pathology, UC Riverside. *⌟ᡜᒌ㸯㜨ᗋኬ㝌࣬⏍࿤⎌㸞

11:15

O-11

Monascus purpureus ࡞࠽ࡄࡾ࢜ࣄẐࢨࢹࣛࢼࣤࡡ⏍ ⏐ㄢ⟿ᅄᏄࡡᶭ⬗よᯊΰỀ೸㞕 1 ࠉᮄୖᾀ 1 ࠉỄ஬ྍ㑳 2ࠉொᖲ༜ஒ 1 㸝1 㜨ኬ࣬⏍∸ᅗ㝷ࢬࠉ2 ࣕ࣢࢝࢞㓬㓕ᢇ◂㸞

11:30

O-12

ⓉⰅ⭁ᮑⳞ⏜ᮮ P450/P450 ࣝࢱࢠࢰ࣭ࢭ⼝ྙࢰࣤ ࣂࢠ㈻ PcCYP17a ࡡᶭ⬗よᯊᚷỀඔ஼ 1ࠉᮿᓧⰟ⨶Ꮔ 1ࠉᗽᮆួႵ 2ࠉ᭯Ἁ❮ 2ࠉᜇᕖ༡ 2ࠉ๪▴༡஄ 1
㸝1 ஐኬ㝌㎨ࠉ2 ࣒ࣜࢨࣔࣤ㸞

11:45

O-13

ⓉⰅ⭁ᮑⳞ Phanerochaete chrysosporium ࡡ㧏✽⣪࠽ ࡻࡦ఩✽⣪᮪௲ୖ࡞࠽ࡄࡾ⣵⬂አࢰࣤࣂࢠ㈻ࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣛࣤࢡᑈᮇ ᐰࠉᚷỀඔ஼ࠉ๪▴༡஄ (ஐኬ㝌࣬㎨)

6

ୌ⯙ㅦⁿ㸝O-14㹳O-19㸞 11 ᭮ 9 ᪝㸝Ề㸞15:30ࠤ17:00
15:30

O-14

Aspergillus oryzae ࢲࣞࢨࢻ࣭ࢭࡡ࢛ࣛࢥ࣏࣭໩ ࡛㓗 Ὡᛮ໩ᶭᵋ
ኣ⏛⨮Ὂኯ㸡㞬ἴ ๙㸡ྚ⏛ Ꮟ 1㸡ୌᓞⱝ἖ 㸝๭౮ኬ㝌࣬⏍∸ᕝ㸡1 ᘧ๑ኬ࣬㎨Ꮥ⏍࿤㸞

15:45

O-15

⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾࣝࢪ࣎ࣤࢪࣝ࢟ ࣭ࣖࣝࢰ࣭ࡡよᯊⴏཋኬ♰ࠉ୷஬ῗୌ᭹ࠉຊ⸠㞖ኃࠉᑚᯐူኰࠉỀ㔕⊓㸝ྞኬ㝌࣬⏍࿤㎨Ꮥ㸞

16:00

O-16

㯔Ⳟࣈࣞࢬࢴࢨࣤࢡࣈࣞࢷ࢓࣭ࢭ (KexB)ࡡḖ᥾࠿⣵⬂ባᵋ㏸࡞ཀྵ࡯ࡌᙫ㡢Ềㆺ἖ 1 㸡᳕ྞᮿᏄ 2, ఫ㔕ඔ᫓ 3, ᒜᙟὊᖲ 2, ῳ㑌๙ᚷ 4, 㜷㒂ᩏᝃ 2, ⏣⏛㞖஄ 5, ୯ᓞబ 2, ஫࿝ົஒ 1  
㸝1 ᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ, 2 ᮶໪ኬ㝌㎨࣬ᚺ⏍໩ࠉ3 㔘Ἁᕝኬࠉ4 ᩺₪ኬ࣬㎨ࠉ5 ⏐⥪◂㸞

16:15

O-17

⣊≟Ⳟ Aspergillus nidulans ࡡ࣐࢛ࢨࣤᵕࢺ࣒࢕ࣤࢅ ᣚࡗ஦ࡗࡡ࢞ࢲࣤྙᠺ㓕⣪
㸝CsmA, CsmB㸞ࡡ⣵⬂හᑻᅹ໩㒂నࠉᶭ⬗Ⓩ┞㛭 㛭౿ࡡよᯊ➁ୖ඼⏠ࠉᇷහ⿩஄ࠉኯ⏛᪺ᚠ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞

16:30

O-18

Putative role of protein kinase C in the phosphorylation and multimerization of the Woroninbody protein, AoHex1, in Aspergillus oryzae
Praveen Rao JUVVADI, Jun-ichi MARUYAMA and Katsuhiko KITAMOTO(Dept. of Biotechnology, Univ. of Tokyo)

16:45

O-19

㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ࢛࣭ࢹࣆ࢒ࢩ࣭ࡡฦ໩ ࠉⓆ ⏍࡫ࡡ㛭୙ⳝ㛣㝧ᚷ㸡ኬ᰷⏛Ꮼ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞

࣎ࢪࢰ࣭Ⓠ⾪ 11 ᭮ 8 ᪝ ዃᩐ␊ྒ 13:30-14:30 ഃᩐ␊ྒ 14:30-15:30
P-1 ࣑࢟㢦㉝࢜ࣄ⑋Ⳟ Fusarium graminearum ࡡⓆⰾ⬂Ꮔ࡞࠽ࡄࡾࣛࣤ㓗໩ࢰࣤࣂࢠ㈻ࡡよᯊ⏍㥎༜ஒ㸡㡪㈙ᬍ஁*㸡㕝ᮄᚥ*㸡H. C. Kistler. **㸡Ⓤ⏣‫‮‬᭹㸝ᒪኬ࣬ᚺ⏍㸡*ᒪኬ࣬⏍࿤ࢬ㸡**Univ. of
Minnesota㸞
P-2 ࢲࣛ⏐ࢹ࣏ࢹᒌ㔕⏍⛸࠾ࡼฦ㞫ࡊࡒ㟸 ⑋ཋᛮ Fusarium oxysporum ࡡฦᏄ⣌⤣よᯊᒱ㒂᪺Ꮔ, ▴ᕖᬰᏄ*, ᕖ㒂┶Ⓡ**, ඡ⋚ᇱୌ㑳***, ᑈᒱ ᚥ, ᭯Ờ ງ 㸝᮶ா㎨ᕝኬ࣬*⌟㣏ဗฦᯊࢬࣤࢰ࣭࣬**࣠ࢨࣤࢹࣤᕗኬ࣬***㫵ཱིኬ㸞P-3 Gibberella fujikuroi Ꮔᄙ⬂Ꮔ࡞࠽ࡄࡾᙟ㈻ࡡฦ㞫࡛ ᇰ㣬୕ΰ࡞ࡻࡾᏄᄙṾᙟᠺㄇᑙᐣᶌຊዄ㸡ᑈᒱ ᚥ㸡᭯Ờ ງ 㸝㎨ᕝኬ࣬㎨㸞
P-4 ࣁ࣭ࢷ࢔ࢨ࣑ࣛࢗⴆ෺⑋Ⳟ Verticillium longisporum ࡡ PCR ᳠ฝ࡛ฦᏄ⣌⤣よᯊ᩟⸠⚵ᠺ㸦㸡᲻ἃ⧂㸦㸡㒿஬Ᏻ㸧㸡⁵ཋᑋᙢ㸧㸡⸠ᮟ┷㸦 㸝㸦᮶Ὂኬ࣬⏍࿤㸡㸧⩄㤷㎨ᢇࢬ࣬⏍⏐⎌ሾ㸞
P-5 ࢗࣛ㢦⅛⑃⑋Ⳟࡡ ClaCWH41 㐿ఎᏄࡢᐙ୹౴ ථ࡞ᚪ㡪࡚࠵ࡽ౴ථჹᏻࡡ⣵⬂ባྙᠺ࡞㛭 ୙ࡌࡾᮿ஬⌦ᜠ࣬ᐋᆀಆᙢ࣬㎱ ඔெ࣬㎱ᒜ ᙪୌ࣬Richard O'Connell࣬Ⓣ▴ཪ⣎㸟࣬஁ಕᗛ஄ 㸝ாᗋኬ㝌㎨࣬
ᒱᒜኬ㎨㸟㸞
P-6 GATEWAY ࢨࢪࢷ࣑ ࢅฺ⏕ࡊࡒ⣊≟Ⳟ⏕㐿ఎᏄ◒ ቪ࣊ࢠࢰ࣭ࡡᵋ⠇㜷㒂 Ṅ㸡Evelyn B. Elegado㸡᭧᰷㍜㞕 㸝໪ኬ㝌㎨࣬ᚺ⏕ⳞᏕ㸞
P-7 ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ DNA ⤄ᥦ࠻ಞᚗ㐿ఎᏄ⩄ࡡよᯊ
Evelyn B. Elegado, 㜷㒂 Ṅ㸡Marites A. Sales㸡ᒷᓧ㜷ᑋ⨶㸡▴஬༐὘*㸡᭧᰷㍜㞕㸝໪ኬ㝌㎨࣬ᚺ⏕ⳞᏕ, *ඔᇳኬ⌦࣬⏍మโᚒ㸞

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P-8 ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ௛╌ჹᙟᠺ࡞ᑊࡌࡾ ࣏ࢠࣞ ࣚ࢕ࢺ⣌ᢘ⏍∸㈻ࡡᙫ㡢Ṃᕖ ἖㸡㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ㸞
P-9 ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡⴝ㠟⣵⬂හ࡚ࡡវ᯹ ິឺ࡛ 㐿ఎᏄⓆ⌟୔⏛ᆀ㈏ྍ࣬㰳⸠៿ୌ㑳࣬㔘᲻ḿᶖ࣬᭯Ờງ࣬ᑈᒱᚥ 㸝㎨ᕝኬ㎨㸞
P-10 ࢕ࢾ࠷ࡵࡔ⑋Ⳟ௛╌ჹᙟᠺ㛭㏻㐿ఎ Ꮔ CBP1 ⏐∸ࡡᑻᅹᛮࡡ᳠ゞᒱᮇಆ㍔㸡㰳⸠៿ୌ㑳*㸡᭯Ờ ງ*㸡ᑈᒱ ᚥ*㸡㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ࣬*㎨ᕝኬ㎨㸞
P-11 ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ ࢭ Hik1 ࡡ᝗ሒఎ㐡⣌ࡡよᯊ᲻⏛┷⣟ 1,2㸡ᮇᒜ㧏ᖶ 1㸡Ꮻఫ⨶ㄵ 2㸡ᕝ⸠ಆ❮ 1 㸝1 ⌦◂࣬୯ኳ◂ࠉ2 ᮶Ὂኬ࣬ᕝ㸞
P-12 ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ஦ᠺฦ᝗ሒఎ㐡⣌ࡡ⎌ሾ ᚺ ➽࡫ࡡ㛭୙ࡡよᯊྡྷᆊ⏜㈏㸡ᮇᒜ㧏ᖶ㸡ᕝ⸠ಆ❮㸝⌦◂࣬୯ኳ◂㸞
P-13 Neurospora crassa ࡡᾈ㏩ᅸ OS ࢨࢡࢻࣜఎ㐡⤊㊪ࡡ ୖὮ࡚โᚒࡈࡿࡾ㐿ఎᏄࡡྜྷᏽ
㔕ཾⲸᯖᏄ㸡ᆊ㔕┷ᖲ, ᕰᕖீኯ, ᮄᮟ┷ 1, ᒜཾຩ 2, ⸠ᮟ┷㸝᮶Ὂኬ࣬⏍࿤, 1 ⌦◂, 2 ᮶Ὂኬ࣬ᕝ㸞
P-14 ࢓࢜ࣂࣤ࢜ࣄࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ-MAP ࢞ࢻ࣭ࢭ⤊㊪࡛ cAMP-PKA ⤊㊪ࡡࢠࣞࢪࢹ࣭ࢠ
ῳ㑌⟿Ꮔ㸦㸡ሲ⃕࠵ࡍࡈ㸦ࠉᆊ㔕┷ᖲ 1㸡ᮄᮟ┷ 2㸡⸠ᮟ┷ 1 㸝㸦᮶Ὂኬ࣬⏍࿤㸡㸧⌦◂࣬୯ኳ◂㸞
P-15 N.crassa ࡡ々ᩐࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ࠿ᾈ㏩ᅸᚺ➽ ࡞㛭୙ࡌࡾሲ⃕࠵ࡍࡈ 1㸡ᒱ⏛᫥౅ 1㸡ⴘྙ์ᖶ 2㸡ᑈ஬᪺✉ 1 㸡ᮄᮟ┷ 2㸡⸠ᮟ┷ 1 㸝㸦᮶Ὂኬ࣬⏍࿤㸡㸧⌦◂࣬୯ኳ◂㸞
P-16 Cryptococcus neoformans ࡡࣀ࢕ࣇࣛࢴࢺᆵ Histidine kinase ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ CnNik1 ࡡᶭ⬗よᯊΰỀපᚠ 1)㸡ῳ㎰ူ 1)㸡ஞ஬ගᙢ 1)㸡Drivinya Antra1)㸡ྚぜၤ 2)㸡⏛୯༐ᑖ 2)㸡ᕖᮇ㐅 1)㸝1㸞༐ⴝኬ࣬┷Ⳟࢬ㸡2㸞ாኬ㝌࣬㎨㸞
P-17 Aspergillus nidulans ࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ㐿ఎᏄ nikAࠉphkAࠉphkB ࡡᶭ⬗よᯊᮿᯐⰃ༡ࠉᒜᓧࡹ࠾ࡽࠉ୷஬ῗୌ᭹ࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞
P-18 ⣊≟Ⳟࡡ Hog1 ᆵ MAPK ࡡὩᛮࡢࢡ࣭ࣜࣈϪࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ࡞ࡻࡖ࡙โᚒࡈࡿࡾ
ྚぜၤ 1 㸡ᑚᓞᾇᖲ 2,3㸡㧏㔕⩇Ꮟ 2 㸡⏛୯༐ᑖ 1 㸝ாኬ㝌㎨࣬1 ᆀ⎌⛁࣬2 ᚺ⏍∸㸡3 ⌟࣬ࢸ࣭ࣖࢠኬ㸞
P-19 ࢓࢜ࣂࣤ࢜ࣄࡡࢨࢡࢻࣜ㐿ఎᏄ ncSCD1ࠉncSCD2 ࡡよᯊᕖᮟ┷ᚷ㸡ୌ▴᫓ᙢ 㸝᮶Ὂኬ࣬⏍࿤⛁Ꮥ㒂㸞
P-20 adenylyl cyclase ✲↓ን␏ࢅᢒᅸࡌࡾ✲↓ን␏మⱩἠᖶኯ 1),ἑහᙹ඼ 1),ᕝ⸠យᏄ 2),ᮟᒜ⫍Ꮔ 1) 1)㛭᮶Ꮥ㝌ኬ࣬ᕝ࣬∸㈻⏍࿤⛁Ꮥ⛁ 2)᪣࢝ࣚࢪ㸝ᰬ㸞
P-21 㯔Ⳟ Aspergillus oryzae ࡢක࡞ᚺ➽ࡊฦ໩ࢅỬᏽࡌࡾ␂ᒜ⌦ᗀ㸡୯℀ᬓ஄, ᵵཾ⿩ḗ㑳, ᭯ᒱ Ꮥ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-22 THE smtA GENE ENCODING A SAM METHYL TRANSFERASE IS REQUIRED FOR NORMAL
DEVELOPMENT OF ASPERGILLUS NIDULANS
Kap-Hoon Han1, Hyo-Jung Kim, Jee Hyun Kim, and Dong-Min Han (Div. Biol. Sci. Wonkwang Univ. Iksan,
570-749, Korea, 1Dept. Pharm. Engin. Woosuk Univ. Wanju, 565-701, Korea)
P-23 Aspergillus oryzae ࡞࠽ࡄࡾ steA ࣓࣌ࣞࢡࡡᶭ⬗よᯊ᲻⏛ᐰெ, ➁හ㐠㞕㸝᮶ா㎨ᕝኬ㝌࣬⏍∸ᕝᏕ㸞

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P-24 Sexuality and asexuality in Aspergillus species
Fabian A. Seymour1, Mathieu Paoletti1, Nanase Yamamoto2, Praveen R. Juvvadi2, Jun-ichi Maruyama2,
Katsuhiko Kitamoto2, David B. Archer1, Paul S. Dyer1 (1School of Biol., Univ. of Nottingham, UK, 2Dept. of
Biotechnol., Univ. of Tokyo)
P-25 㯜㯔Ⳟ⏍⫩Ⳟమහࣈࣞࢷ࢛࣭࣑ࡡよᯊ♼ኯ㑳㸡ࢡ࢙ࣤ࣬ࢤࣤ࣬ࣀ㸡➁හ㐠㞕 㸝᮶ா㎨ᕝኬᏕ࣬ᚺ⏕⏍∸⛁Ꮥ⛁㸞
P-26 ⣊≟Ⳟ࡞࠽ࡄࡾ APase 㐿ఎᏄࡡኣᵕᛮ
ᒱᮇ⥜Ꮔ㸡➁හ㐠㞕 㸝᮶ா㎨ᕝኬᏕ࣬㎨࣬ᚺ⏍⛁㸞
P-27 ⣊≟Ⳟ Aspergillus nidulans ࡡࣈࣞࢬࢴࢨࣤࢡ㓕⣪㐿ఎᏄ kexB ◒ቪᰬࡡよᯊୌᰏಆ௒ 1, ཿᕖ೸ኯ㑳 1, Ềㆺ἖ 2, ⸠ᒱᬓ์ 1, ᚠᒱ᪸ᩝ 2, ஫࿝ົஒ 2, 㜷㒂ᩏᝃ 1 (᮶໪ኬ㝌㎨࣬1 ᚺ⏍⛁, 2 ⏍∸⏐ᴏ๭⏍)
P-28 ⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾ class III ࢞ࢲࣤฦよ㓕⣪ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄࡡᶭ⬗よ ᯊᒜᓧᬍ୓㸡ᇷහ⿩஄ࠉኯ⏛᪺ᚠ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-29 Aspergillus tamarii ⏜ᮮࡡ niaD 㐿ఎᏄࡡᵋ㏸
ᮄᮟኣỜ 1 ᴃᮇ៿ୌ 1 ໪ᮇ์⾔ 2  㕝ᮄ⪵ 1 ᯵ᮄ㇇ 1 㸝1 ≺Ἢ࣬㣏⥪◂, 2 យ▩⏐ᢇ◂࣬㣏ဗᕝᢇ㸞
P-30 Insights into RIP and DNA methylation in the Aspergillus section Flavi complex
Heather A. Lee, Maria Dolores Montiel and David B. Archer㸝 Institute of Genetics, School of Biology,
University of Nottingham, University Park, Nottingham, NG7 2RD 㸞
P-31 㯔Ⳟࡡ G ࢰࣤࣂࢠ㈻භᙲᆵ cAMP ུᐖమ┞ྜྷ㐿ఎᏄ㕝ᮄ ⪵㸡➁ㆺ༡Ꮔ, ᮄᮟኣỜ, ᮿୖ┷⏜⨶, ᴃᮇ៿ୌ, ᯵ᮄ ㇇ 㸝㣏⥪◂㸞
P-32 Dikaryon ࡡฦ⿛࡞࠽ࡄࡾࢠࣚࣤࣈࡡណ⩇ᒱᓧᏏ᫆㸡୸⩒ಞ㌗ 㸝࠾ࡍࡈ DNA ◂✪ᡜ࣬᯹Ⰵమᶭ⬗㡷ᇡ➠ୌ㸞
P-33 smuH501, a second mutation found in the original uvsH77 mutant strain that involved in DNA replication
checkpoint control in Aspergillus nidulans
Mee-Jeong Cha, Sun-Hee Noh, Nak-Jung Kwon, and Suhn-Kee Chae (Dept. Biochem. and Biomed RRC,
Paichai Univ. Daejeon, 302-735 Korea)
P-34 ⺧කࢰࣤࣂࢠᵾㆉࢅ⏕࠷ࡒ Aspergillus nidulans ࢰࣤࣂࢠ㈻ࡡ⣵⬂࿔᭿ንິࡡࣚ࢕ࣇ࢕࣒࣭ ࢩࣤࢡ㸯 II ᆵ topoisomerase ࢅ୯ᚨ࡞1⣒ ᭰ᾀ㸡ᕖᓃ⨶౅㸡ⳝờ㡝ᙢ㸡2Berl R. Oakley㸡3 ᇷᑹူஒ㸝1 ྞཿᒁኬ㝌༈⣌࣬ฦᏄᵾⓏ㸡2 ࢛ࣀ࢕࢛ᕗ❟ኬ࣬ฦᏄ㐿ఎ㸡3 ᚠᓞኬ㝌࣬ࣉࣜࢪࣁ࢕࢛㸞

࣎ࢪࢰ࣭Ⓠ⾪ 11 ᭮ 9 ᪝ ዃᩐ␊ྒ 13:30-14:30 ഃᩐ␊ྒ 14:30-15:30
P-35 ࢨ࢕ࢰࢢ(Lentinula edodes)ࡡș-1,3-ࢡࣜ࢜ࢻ࣭ࢭࡡよᯊᆊᮇ⿩ୌ㸡Ễ஬ົ㸡୯ฝၤᏄ㸡ఫ⸠ฺḗ 㸝ᒷᡥ⏍ᕝ◂㸞
P-36 ࢨ࢕ࢰࢢࡡᏄᐁమᙟᠺ㐛⛤࡞࠽࠷࡙≁␏Ⓩ ࡞ Ⓠ⌟ࡌࡾ㐿ఎᏄ⩄ࡡ༟㞫࡛よᯊᐋᓧᏭᑑࠉ୯ᮟ㞖သࠉ㤷ሔᓧົᙢ㸝᲻ᯐ⥪◂㸞

9

P-37 ᢰᏄⳞ Phanerochaete chrysosporium ࠿᭯ࡌࡾ P450 㐿ఎᏄࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣛࣤࢡ
୯ᮟ▩ᜠ 1ࠉᗽᮆួႵ 2ࠉᖲሪᐃ༡ 1 ࠉᒜ⏛ಞྒྷ 3ࠉばἡᨳ஦୔ 3 ࠉ᭯Ἁ ❮ 2ࠉᜇᕖ ༡ 2ࠉ๪▴༡஄ 1
㸝1 ஐኬ㝌㎨ࠉ2 ࣒ࣜࢨࣔࣤ⏍㈠◂ࠉ3JST ๪▴ࣈࣞࢩ࢘ࢠࢹ㸞
P-38 ᢰᏄⳞ Phanerochaete chrysosporium ࡞࠽ࡄࡾࢬ࣭ࣜࣞࢪฦよ㓕⣪㐿ఎᏄⓆ⌟ᚺ➽࡞㛭ࡌࡾ ◂✪㕝ᮄୌྍࠉ஫༎ᔋᅽ᪝Ꮔࠉ㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞P-39 ᢰᏄⳞ Phanerochaete chrysosporium ⏜ᮮࢹࣝࣀ࣭ࣞࢪຊࣛࣤ㓗ฦよ㓕⣪ࡡᶭ⬗よᯊᖲ▴ḿ⏠ࠉ஫༎ᔋᅽ᪝Ꮔࠉ㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞P-40 ᢰᏄⳞ Phanerochaete chrysosporium ࠿⏍⏐ࡌࡾ⢶㈻ຊỀฦよ㓕⣪ࣆ࢒࣐࣭ࣛ74 ࢞ࢨࣞࢡࣜ࢜ࢻ࣭ࢭࡡᶭ⬗よᯊ
▴⏛༜ஒ 1ࠉຊ἖ఫᖲ 1ࠉ஫༎ᔋᅽ᪝Ꮔ 1ࠉ▦㏛ග㑳 2ࠉ᪝ྚ࠵ࡷᏄ 2ࠉ୔▴ Ꮽ 2ࠉ㩢ᓞḿᾀ 1㸝1 ᮶ኬ㝌࣬㎨⏍⛁ࠉ2 ⏐⥪◂㸞
P-41 ᢰᏄⳞ Phanerochaete chrysosporium ࠿⏍⏐ࡌࡾ⢶㈻ຊỀฦよ㓕⣪ࣆ࢒࣐࣭ࣛ1 ࡞ᒌࡌࡾș-ࢡࣜࢤࢨࢱ࣭ࢭࡡᶭ⬗よᯊሪ⏛๙ኃ 1ࠉ஦༎㌲᜾㔓 1ࠉྚ⏛ㄌ 2 ࠉఄಘ㐅▦ 1ࠉ஫༎ᔋᅽ᪝Ꮔ 1ࠉ㩢ᓞḿᾀ 1㸝1 ᮶ኬ㝌࣬㎨⏍⛁ࠉ2
㣏⥪◂㸞
P-42

DNA ࡡ㟸≁␏Ⓩቌᖕࢅฺ⏕ࡊࡒᮄᮞ⭁ᮑⳞ ࡡ㧏វᗐ᳠ฝ࿰⏛᭰Ꮔ㸡ຊ἖ఫᖲ㸡஫༎ᔋᅽ᪝Ꮔ㸡㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞P-43 㯔Ⳟ࡞࠽ࡄࡾ᩺ぞࡡ⢶㙈㛭㏻Ὡᛮࢰࣤࣂࢠ ࡡ ᥀⣬⋚㔕Ꮟୌ㸡ᑈᯐ㟱ᐃ㸡ఫ⸠ཪ⣎㸡ᇷ㉲Ờ⨶*㸡▹ᕖ⨶ఫ⥬㸡ᒜ᰷೒Ꮔ㸡⏣⏛㞖஄㸝⏐⥪◂㹺⏍∸ᶭ⬗㸡*᪝ኬ࣬⏍⏐ᕝᏕ㸞

P-44 A. oryzae ࡞࠽ࡄࡾ N-⤎ྙᆵ⢶㙈࡛┞பష⏕ࡌࡾࢰ ࣤࣂࢠ㈻ࡡࢪࢠ࣭ࣛࢼࣤࢡ
୷ᒜ₮ୌ 1ࠉᮿᑹୌ㑳 2ࠉ௿⸠ᖶᠺ 2 ࠉ໪ᮇົࡥࡆ 1  㸝1 ᮶ኬ㝌㎨⏍⛁࣬ᚺ⏍ᕝࠉ2 ⌦◂㸞
P-45 㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ ER-Golgi మ㛣ࡡ㍲㏞࡞㛭ࢂࡾ SNARE ࡡᑻᅹよᯊ⏛὾⥜Ꮔ㸡ಲ㒌ᑑᏳ, ḿ㊪ῗஒ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-46 㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ post-Golgi ㍲㏞࡞㛭ࢂࡾ SNARE ࡡᑻᅹよᯊಲ㒌ᑑᏳ㸡ḿ㊪ῗஒ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-47 㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ᥃ྙࣆ࣓࢘ࣞࣤ๑㥉మ 㐿ఎᏄ AoppgA ࡡᶭ⬗よᯊ
ᒜᮇ୏℡ࠉJUVVADI Praveen Raoࠉ୷ᒜ₮ୌࠉ໪ᮇົࡥࡆ (᮶ኬ㝌㎨⏍⛁࣬ᚺ⏍ᕝ)
P-48 ࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪ࡞ࡻࡾ㯔Ⳟ A. oryzae ࢅ⏕࠷ࡒࣃࢹࣛࢯࢲ࣭࣑⏍⏐㔖ࡡቌຊ
ῳ㎰ὀ♰㸡㔘㗋᮵㸡୷ᒜ₮ୌ㸡Praveen Rao Juvvadi㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-49 㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ 2 ࡗࡡฦἢᆵ࣌ࢪ࣌ࣛ ࣂ࣭ࢭ A2 ࡡᢘమࢅ⏕࠷ࡒᑻᅹよᯊ࡛ RT-PCR ࡞ ࡻࡾⓆ⌟よᯊ୯℀ᬓ஄㸡໪ᮇົࡥࡆ㸡᭯ᒱ Ꮥ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞
P-50 㯔Ⳟ A. oryzae ࢅ⏕࠷ࡒࢹࣚࣆࢡ࣬ࣂࣆࣝࢠࢲ ࣤࡡ⏍⏐➪ᰡᚷಕ, 㤷ᶣ⏜౅, ୷ᒜ₮ୌ, ᭯ᒱᏕ, ໪ᮇົࡥࡆ (᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ)
P-51 㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ Aovps24 (class E vps)㐿ఎᏄࡡ༟㞫࡛よᯊ㎦ᕦᬏ⣎㸡ⳝ㛣㝧ᚷ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ (᮶ኬ㝌㹺㎨⏍⛁㹺ᚺ⏍ᕝ)P-52 㯔Ⳟ A. oryzae ࡡ III ᆵ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㐿ఎᏄࡡⓆ⌟࡛ᶭ⬗よᯊເࠎᗛ௥ 1㸡⸠஬ ໌ 2㸡Praveen Rao Juvvadi1㸡ᾇ⩹ሪ㇇ 2㸡໪ᮇົࡥࡆ 1㸝1 ᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ,
2᮶ኬ㝌࣬ⷾ㸞

10

P-53 Penicillium luteo-aurantium ࡡ᩺ぞ࣎ࣛࢢࢰ࢕ࢺྙᠺ 㓕⣪㐿ఎᏄࡡࢠ࣭ࣞࢼࣤࢡ࡛ᶭ⬗よᯊఫ㔕⿩ᇱࠉ▴஬⤦⨶ࠉ⸠஬ ໌ࠉᾇ⩹ሪ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞
P-54 ⣊≟Ⳟⰶ㤮᪐࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ࡡࢠࣚ ࢕ ࢭࣤࢦ࢕ࢠ࣭ࣚࢭࢺ࣒࢕ࣤᐋᓧ ಆࠉ⸠஬ ໌ࠉᾇ⩹ሪ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞
P-55 ⰶ㤮᪐⧖㏁ࡊᆵࢰ࢕ࣈϨ࣎ࣛࢢࢰ࢕ࢺྙᠺ 㓕 ⣪ࡡⓆ⌟࡛ᶭ⬗よᯊ᲻ཾᬓ⨶㸡 ⸠஬ ໌㸡 ᾇ⩹ሪ㇇ 㸝᮶ኬ㝌ⷾ㸞
P-56 ⣊≟Ⳟ A. nidulans ࢓࣐࣭ࣚࢭ㐿ఎᏄ⩄ࡡⓆ⌟ ࣈࣞࣆ࢒࢕ࣛࣤࢡ
୯ᮟ㈏㸡∶⏛ᬓ⿩㸡ຊ⸠㞖ኃ㸡ᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞
P-57 㯔Ⳟ࢞ࢨࣚࢻ࣭ࢭ࣬ࢬ࣭ࣜࣚࢭㄇᑙᅄᏄ AoXlnR ࡡ㌹෕โᚒᶭᵋ
㔕ཾ♰ྒྷ㸡⏛୯ᑋᇱ㸡ຊ⸠㞖ኃ㸡ᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞
P-58 㯔Ⳟ CCAAT ⤎ྙᅄᏄඁເ㜴ᐐᆵን␏ࢦࣇࣗ ࢼࢴࢹࡡよᯊ㸯โᚒࡈࡿࡾ㐿ఎᏄ⩄ࡡ⥑⨮Ⓩよᯊ࡫ࡡฺ⏕ᮙᒜ⣟ஒࠉ㧏ᶣ᪺⌌ࠉྙ⏛⚵▦ࠉᑚᯐူኰࠉຊ⸠㞖ኃ (ྞኬ㝌࣬⏍࿤㎨࣬⏍∸ᶭᵋ)
P-59 㯜㯔Ⳟ Aspergillus oryzae ࡡ HSP30 㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭ࡡ Deletion よᯊᮿୖ┷⏜⨶ࠉ㕝ᮄ ⪵ࠉᴃᮇ៿ୌࠉ᯵ᮄ ㇇ 㸝㸝≺㸞㣏⥪◂㸞
P-60 㯔Ⳟ Aspergillus oryzae ࡡ఩Ὼㄇᑙࣈ࣓࣭ࣞࢰ࣭ࡡ༟㞫஁⏛༡ඔ㸡ላᾀᏄ㸡⛑Ὂ஦㸡Ꮽ㒂ᗛ஁㸝᭮᰿෗⥪◂㸞
P-61 㯔Ⳟ࡛㓕ẍ࡞࠽ࡄࡾ㐿ఎᏄⓆ⌟ࡡẒ㍉よᯊᒜ᰷೒Ꮔ㸡ᑈᯐ㟱ᐃ㸡ఫ㔕ඔ᫓*㸡㧏℡஁⨶Ꮔ㸡ኬ⟺ಘୌ*ࠉ⏣⏛㞖஄ 㸝⏐⥪◂࣬*㔘Ἁᕝኬ㸞
P-62 Carbon and nitrogen repression of the expression of citA gene encoding citrate synthase in Aspergillus
nidulans
Pil Jae Maeng* and Yeong Man Yu (Dept. of Microbiology, Chungnam National Univ., Daejeon 305-764,
Korea)
P-63 RNAi ࡞ࡻࡾ㯑Ⳟ(Aspergillus oryzae)㐿ఎᏄࢦ࢕ࣝࣤ ࢨࣤࢡ⣌ࡡ㛜Ⓠᒜ⏛ ಞ㸡ᆊᮇ࿰ಆ㸡⚽⏛ ಞ 㸝㒿⥪◂㸞
P-64 ࢠ࢙ࣤ㓗⏍⏐⣊≟Ⳟ Aspergillus niger ࡞࠽ࡄࡾ ⥫Ⰵ⺧කࢰࣤࣂࢠ㈻ࢅ࣏࣭࣭࡛࢜ࡊ࡙⏕࠷ ࡒ
alternative oxidase 㐿ఎᏄ㸝aox1㸞ࡡⓆ⌟よᯊ
ᑚἑ㈶ྍ㸡᭱㒂㈏⃀, ᮄ㔕㑝ჹ㸡᱊ᮟකኯ㑳 㸝᪡ኬ࣬⌦ᕝ࣬ᚺ໩㸞

11

S-1
㯔ⳞⓆ⌟⏕ࣈࣚࢪ࣐ࢺࡡ㉰㎷㏷షᠺἪ࡛ࡐࡿࢅ⏕࠷ࡒࣆࢡࢰࣤࣂࢠ㈻ࡡ⏍⏐᭯ᒱ Ꮥ㸝᮶ாኬᏕኬᏕ㝌㎨Ꮥ⏍࿤⛁Ꮥ◂✪⛁ᚺ⏕⏍࿤ᕝᏕᑍᨯ㸞ࡢࡋࡴ࡞࣎ࢪࢹࢣࢿ࣑᫤௥ࡡ฽ᮮ࡞ఔ࠷ࠉ◂✪ᮞᩩ࡛ࡊ࡙౐⏕ࡌࡾ㧏ဗ㈻࡝ࢰࣤࣂࢠ㈻ࠉ࠵ࡾ࠷ࡢ༈ⷾ⏕㏭࡝
࡜ࡡ௛ຊ౮ೋࡡ㧏࠷᭯⏕ࢰࣤࣂࢠ㈻ࢅኬ㔖࡞ࡐࡊ࡙㎷㏷࡞⏍⏐ࡌࡾᢇ⾙ࡡ㐅Ṅ࠿ᙁࡂ᭻ࡱࡿ࡙࠷ࡾࠊኬ
㔖࡞ࠉᏭ౮࡞ࠉ࠾ࡗ▯᫤㛣࡚ࢰࣤࣂࢠ㈻ࢅ⏍⏐ࡌࡾ࡞ࡢ᚜⏍∸ᐙ୹࠿㐲ࡊ࡙࠷ࡾ࠿ࠉ୯࡚ࡵ⣊≟Ⳟࡢ㓕ẍ࡝࡜࡞Ẓ࡬࡙ࡢࡾ࠾࡞ኣ㔖ࡡࢰࣤࣂࢠ㈻ࢅ⣵⬂አ࡞ฦἢࡌࡾ⬗ງࢅᣚࡗࡆ࡛࠾ࡼࠉ㓕⣪๠࡝࡜ࡡ⏍⏐࡞ᗀࡂ⏕࠷ࡼࡿ࡙ࡀࡒࠊ≁࡞㯔Ⳟ Aspergillus oryzae ࡢᠻ࠿ᅗࡡఎ⤣Ⓩ࡝㔂㏸⿿ဗ࡚࠵ࡾ㒿ࠉ࿝ძࠉ㓲ἔ࡝࡜࡛࠷ࡖࡒᵕࠍ࡝㣏ဗࡡ⿿㏸࡞ 1000 ᖳ௧୕࡞ࢂࡒࡖฺ࡙⏕ࡈࡿ࡙ࡀࡒ⤊㥺࠾ࡼࠉெమ࡞࡛ࡖ࡙Ꮽධ
ᛮࡡ㧏࠷ᐙ୹࡛ࡊ࡙ヾࡴࡼࡿ࡙࠽ࡽࠉࡐࡡ㧏࠷ࢰࣤࣂࢠ㈻ฦἢ⏍⏐⬗ງࢅฺ⏕ࡊ࡙㣏ဗ࣬໩ᠺဗ࣬༈ⷾ
ဗྡྷࡄࡡ㓕⣪๠࠽ࡻࡦ௥ㅨ໩ྙ∸࠿⏍⏐ࡈࡿᅗ㝷Ⓩ࡞ᗀࡂὮ㏳ࡊ࡙࠷ࡾࠊ
㯔Ⳟࢅ⏕࠷࡙␏⛸ࢰࣤࣂࢠ㈻ࠉ≁࡞ฦἢࢰࣤࣂࢠ㈻ࢅ⏍⏐ࡌࡾ㝷࡞ࡢࠉୌ⯙࡞ᙁງ࡝ࣈ࣓࣭ࣞࢰ࣭ࢅ
⏕࠷ࠉࢡࣜࢤ࢓࣐࣭ࣚࢭ࡝࡜㧏Ⓠ⌟ࡌࡾ࢞ࣔࣛ࢓ࢰࣤࣂࢠ㈻࡛ࡡ⼝ྙࢰࣤࣂࢠ㈻࡛ࡊ࡙Ⓠ⌟ࡈࡎࡾࡆ࡛
࠿ኣ࠷ࠊࡱࡒⓆ⌟ࡡ࣍ࢹࣜࢾࢴࢠ࡛࡝ࡾᵕࠍ࡝さᅄࢅᤴ㝎ࡌࡾࡒࡴࠉᐙ୹࡞ᑊࡊ࡙ࢨࣔ࣋ࣞࣤࡡ㧏Ⓠ⌟
ࡷࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄࡡ◒ቪ࡝࡜ࡡᨭንࢅຊ࠻ࡾࡆ࡛ࡵ⾔ࢂࡿࡾࠊࡆ࠹ࡊࡒ᧧షࢅຝ⋙ࡻࡂ⾔࠹ࡒࡴ࡞ࡢࠉࡐࡿࡼ࡞⏕࠷ࡾኣᩐࡡࣈࣚࢪ࣐ࢺࢅ㎷㏷࡞ᵋ⠇ࡌࡾᚪさ࠿࠵ࡾ࠿ࠉโ㝀㓕⣪࡛ࣚ࢕ࢣ࣭ࢨࣘࣤཬᚺ
ࢅ⏕࠷ࡾᚉᮮࡡࣈࣚࢪ࣐ࢺᵋ⠇Ἢ࡚ࡢࠉ≁࡞ኣ⛸㢦ࡡⓆ⌟ࣈࣚࢪ࣐ࢺᵋ⠇ࢅ୩⾔ࡊ࡙⾔࠹㝷࡞ࡊࡣࡊࡣ
ᅏ㞬࠿ఔ࠹ࠊ
㎾ᖳࠉ㐿ఎᏄࡡ㎷㏷࡝ࢠ࣭ࣞࢼࣤࢡࢅ⾔࠹ࡒࡴࡡᢇ⾙࡛ࡊ࡙ MultiSite Gateway ࢷࢠࢿࣞࢩ࣭࠿㛜Ⓠࡈ
ࡿࡒࠊࡆࡿࡢλࣆ࢒࣭ࢩࡡ⤄ᥦ࠻ཬᚺࢅฺ⏕ࡌࡾࡆ࡛࡞ࡻࡽࠉࣚ࢕ࢣ࣭ࢨࣘࣤཬᚺࢅ⾔࠹ࡆ࡛࡝ࡂ々ᩐࡡ DNA ᩷∞ࢅୌࡗࡡ࣊ࢠࢰ࣭࡞⤄ࡲ㎲ࡳࡆ࡛ࢅྊ⬗࡞ࡌࡾࢨࢪࢷ࣑࡚࠵ࡾࠊᠻࠍࡢࡆࡡࢨࢪࢷ࣑ࢅ⏕࠷࡙㯔Ⳟ⏕ࡡⓆ⌟࣊ࢠࢰ࣭࠽ࡻࡦᐙ୹ᨭን⏕ࡡ⛸ࠍࡡࣈࣚࢪ࣐ࢺࢅᴗࡴ࡙ᐖ᪾࡞ష⿿ࡌࡾࡆ࡛ࡡ࡚ࡀ
ࡾ࣊ࢠࢰ࣭⣌ࡡ㛜Ⓠࢅ⾔ࡖࡒࠊᮇⓆ⾪࡚ࡢࡢࡋࡴ࡞ࡐࡡᴣㄕࢅ⾔࠷ࠉ⤾࠷࡙ࡐࡿࢅฺ⏕ࡊࡒ␏⛸ࢰࣤࣂ
ࢠ㈻ࡡ⏍⏐ᐁ㥺࡞㛭ࡊ࡙⣺௒ࡊࡒ࠷ࠊ
A. oryzae ࡞࠽ࡄ ࡾ᩺ぞ Ⓠ⌟࣊ ࢠࢰ࣭ ⣌ࡡᵋ ⠇࡛ ࡐࡿࢅฺ⏕ࡊࡒ࣐ࢹࢤࣤࢺࣛ࢓ ࡡྊっ ໩
Multisite Gateway ࢨࢪࢷ࣑࡚ࡢࡐࡿࡑࡿ 5'-ࠉ୯ኳࠉ࠽ࡻࡦ 3'-࢙ࣤࢹ࣭ࣛࢠ࣭ࣞࣤ (EC) ୕࡞ࢠ࣭ࣞࢼ
ࣤࢡࡈࡿࡒ 3 ࡗࡡ DNA ᩷∞ࢅᏽࡴࡼࡿࡒ㡨ᗆ࡛᪁ྡྷᛮ࡚ 1 ࡗࡡ࣊ࢠࢰ࣭࡞⛛ࡌࡆ࡛࠿࡚ࡀࡾࠊᠻࠍࡢ
5'-EC ࡛ࡊ࡙ࣈ࣓࣭ࣞࢰ࣭࠵ࡾ࠷ࡢࣈ࣓࣭ࣞࢰ࣭㸠࢞ࣔࣛ࢓ࢰࣤࣂࢠ㈻㐿ఎᏄࢅ᭯ࡌࡾࣈࣚࢪ࣐ࢺࠉࡱ
ࡒ୯ኳ࠽ࡻࡦ 3'-EC ࡛ࡊ࡙ N-࠵ࡾ࠷ࡢ C-ᮆ❻⼝ྙࢰࢡ㐿ఎᏄࢅ᭯ࡌࡾࣈࣚࢪ࣐ࢺࢅ⏕ណࡊࡒࠊරమⓏ࡞
ࡢࠉࣈ࣓࣭ࣞࢰ࣭⏕࡛ࡊ࡙ 3 ⛸ (amyBࠉthiA ࠽ࡻࡦ pgkA)ࠉࣈ࣓࣭ࣞࢰ࣭㸠࢞ࣔࣛ࢓ࢰࣤࣂࢠ㈻⏕࡛ࡊ࡙ 2 ⛸ (amyB ࠽ࡻࡦ glaA) ࡡ㒼าࢅᣚࡗ 5'-EC ࢅࠉࡱࡒ N-࠵ࡾ࠷ࡢ C-ᮆ❻⼝ྙࢰࢡ⏕࡛ࡊ࡙ EGFPࠉDsRed2ࠉGSTࠉHA-His6 ࡡ྘㒼าࢅᣚࡗ୯ኳ࠵ࡾ࠷ࡢ 3'-EC ࢅྙゝ⣑ 10 ⛸㢦ᵋ⠇ࡊࡒࠊࡆࡿ࡞ࡻࡽࠉࡆ
ࡿࡼ EC ࡛┘Ⓩ㐿ఎᏄࢅ᭯ࡌࡾ୯ኳ
࠵ࡾ࠷ࡢ 3'-EC ࡛ࢅ㐲ᙔ࡞⤄ࡲྙࢂࡎࡾࡆ࡛࡚ᩐ༎⛸㢦ࡡⓆ⌟ࣈࣚࢪ࣐ࢺࢅ⠾౼࡞ᚋࡾࡆ࡛࠿࡚ࡀࡾࡻ࠹࡞
࡝ࡖࡒࠊࡱࡒࠉࡆࡿࡼ࡛ࡢื࡞㐽ᢝ࣏࣭࣭࢜ࡡࡲࢅᣚࡗ୯ኳ EC ࡵ⏕ណࡊࠉ㐿ఎᏄ◒ቪ⏕ࣈࣚࢪ࣐ࢺࢅᐖ᪾
࡞ష⿿࡚ࡀࡾࡻ࠹࡞ࡊࡒࠊ
ᵋ⠇ࡊࡒ㯔Ⳟ⏕࣊ࢠࢰ࣭ࢨࢪࢷ࣑
ࡡ᭯ຝᛮࢅ☔ヾࡌࡾࡒࡴࠉ࣐ࢹࢤࣤࢺࣛ࢓ྊっ໩⏕ࡡࣈࣚࢪ࣐ࢺࢅష⿿
ࡊࡒࠊ
5'-EC ࡛ࡊ࡙ amyB ࠵ࡾ࠷ࡢ thiAࣈ࣓࣭ࣞࢰ࣭ࠉ୯ኳ EC ࡛ࡊ࡙࣐ࢹࢤ
ࣤࢺࣛ࢓ᑻᅹ໩ࢨࢡࢻࣜࢅྱࡳ㯔Ⳟ
citrate synthase ┞ྜྷ㒼าࡡ N ᮆ❻ഁ㒂ฦࠉࡱࡒ 3'-EC ࡛ࡊ࡙ EGFP ࠵ࡾ࠷ Multisite Gateway ࢅฺ⏕ࡊࡒ㯔ⳞⓆ⌟⏕ࣈࣚࢪ࣐ࢺష⿿ࢨࢪࢷ࣑㻃ࡢ DsRed2 ࢅ㐽ᢝࡊࡒࠊᚋࡼࡿࡒⓆ⌟ Center-࠽ࡻࡦ 3‘-EC ࡞ࡗ࠷࡙ࡢࡆࡡ㏣ࡡ⤄ࡲྙࢂࡎࡡࣈࣚࢪ࣐ࢺࡵష⿿ࡊࡒ
⏕ࣈࣚࢪ࣐ࢺࢅ㯔Ⳟ࡞ᑙථࡊࠉ⺧ක㢟᚜㙶࡚ふᐳࡊࡒ࡛ࡆࢀࠉ࣐ࢹࢤࣤࢺࣛ࢓࡛⩻࠻ࡼࡿࡾኣᩐࡡࢲࣖ

12࣭ࣇ≟ᵋ㏸࠿ࠉⓆ⌟ࢅㄇᑙࡌࡾ᮪௲ୖ࡚ࡡࡲヾࡴࡼࡿࡒࠊࡆࡿࡼࡢ࠷ࡍࡿࡵ Mitotracker 㝟ᛮ࡚࠵ࡖࡒࡆ
࡛࠾ࡼࠉ᩺ぞࢨࢪࢷ࣑࠿ゝ⏤㏳ࡽᶭ⬗ࡌࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊ᩺ぞࢨࢪ ࢷ࣑ࢅ ฺ⏕ࡊ ࡒ㨮㢦 ⏍⌦Ὡ ᛮࢰࣤ ࣂࢠ ㈻ࡡ⏍⏐⤾࠷࡙᮶ாኬᏕ௛ᒌỀ⏐ᐁ㥺ᡜ࡛ࡡභྜྷ◂✪࡛ࡊ࡙ࠉ᩺ぞࢨࢪࢷ࣑ࢅ⏕࠷࡙ࢹࣚࣆࢡ⏜ᮮࡡࣝࢠࢲࣤ
࡚࠵ࡾࣂࣆࣝࢠࢲࣤ (FrPl) ࡡ⏍⏐ᐁ㥺ࢅ⾔ࡖࡒࠊ5'-EC ࡛ࡊ࡙ amyB ࣈ࣓࣭ࣞࢰ࣭࠽ࡻࡦ࢞ࣔࣛ࢓ࢅࠉ
3'-EC ࡛ࡊ࡙よᯊ࠽ࡻࡦ⢥⿿࡞⏕࠷ࡾ HA-His6 ࢰࢡࢅ㐽ᢝࡊࠉFrPl ࡢ୯ኳ EC ࡛ࡊ࡙ㄢ⿿ࡊࡒࠊࡐࡡ㝷ࠉ
amyB ࡛ FrPl ࡛ࡡ㏻⤎㒂࡞ࡢ Kex2 ᵕࣈࣞࢷ࢓࣭ࢭ࡞ࡻࡾヾㆉ㒼าࢅ᣼ථࡊࡒࠊᚋࡼࡿࡒࣈࣚࢪ࣐ࢺ࡚
ᙟ㈻㌹ᥦࡊࡒ㯔Ⳟࡡᇰ㣬୕ΰࢅᢘ HA ᢘమࢅ⏕࠷࡙よᯊࡊࡒ࡛ࡆࢀࠉ┘Ⓩ࡛ࡌࡾ FrPl::HA-His6 ⼝ྙࢰࣤࣂࢠ㈻ࡡ஢᝷ฦᏄ㔖ࢅᣚࡗࣁࣤࢺ௧አ࡞ࡵࠉ࢞ࣔࣛ࢓ࢰࣤࣂࢠ㈻࡛㏻⤎ࡊࡒࡱࡱࡡ≟ឺ࡛⩻࠻ࡼࡿࡾࣁ
ࣤࢺࡵヾࡴࡼࡿࡒࠊࡐࡆ࡚ Kex2 ヾㆉ㒼าࡡ├ᚃ࡞ࢡࣛࢨࣤࢅ 3 ṟᇱᑙථࡊࡒࣈࣚࢪ࣐ࢺࢅྜྷᵕ࡞ࡊ࡙
ᵋ⠇ࡊࠉࣈࣞࢷ࢓࣭ࢭ࡞ࡻࡾว᩷ຝ⋙࠿୕᪴ࡌࡾ࠾࡜࠹࠾ࢅ᳠ゞࡊࡒࠊࡐࡡ⤎ᯕࠉ㧏ฦᏄ㔖ࡡࣁࣤࢺ࠿
ᾐ኶ࡊࠉ࡮࡯Ᏸධ࡞ว᩷ࡈࡿࡒࣁࣤࢺࡡࡲ࠿ヾࡴࡼࡿࡒࠊࡆࡿࢅ Ni2+࣑࢜ࣚࢅ⏕࠷࡙⢥⿿ࡊࠉ࢓࣐ࢿ㓗㒼าࡡよᯊࢅ⾔ࡖࡒ⤎ᯕࠉFrPl ࡡ N ᮆ❻㒼า࠿᳠ฝࡈࡿࠉࠉ┘Ⓩ࡛ࡌࡾ⼝ྙࢰࣤࣂࢠ㈻࠿ฦἢ⏍⏐ࡈࡿࡒࡆ࡛࠿☔ヾࡈࡿࡒࠊ⢥⿿ࡊࡒ⤄ᥦ࠻ FrPl ࢅ⏕࠷࡙⾉⌣ฆ㞗Ὡᛮ࠽ࡻࡦࣝࢠࢲࣤὩᛮࢅㄢ࡬ࡒ࡛ࡆࢀࠉ୦Ὡᛮ࡛ࡵヾࡴࡼࡿࡒࠊ⌟ᅹࠉࡈࡼ࡞ኬ㔖ࡡࢦࣤࣈࣜࡡㄢ⿿ࢅモࡲ࡙࠷ࡾࠊ
௧୕ࡡࡻ࠹࡞ࠉᮇ◂✪࡚ᵋ⠇ࡈࡿࡒࢨࢪࢷ࣑ࢅ⏕࠷ࡾࡆ࡛࡚㯔Ⳟ⏕ࡡⓆ⌟࣊ࢠࢰ࣭ࢅᴗࡴ࡙ᐖ᪾࡞ష
⿿ࡌࡾࡆ࡛࠿࡚ࡀࡾࡻ࠹࡞࡝ࡖࡒࠊᐁ㝷ࠉᙔ◂✪ᐄ࡚ࡢࡆࡿࢅ⏕࠷࡙ᩐኣࡂࡡࣈࣚࢪ࣐ࢺ࠿ష⿿ࡈࡿ࡙
࠽ࡽࠉ◂✪ࡡ㎷㏷໩࠿ᠺࡈࡿ࡙࠷ࡾࠊᮇࢨࢪࢷ࣑ࡡὩ⏕ࡢㄢ⿿ࡌࡾࣈࣚࢪ࣐ࢺࡡ⛸㢦࠿ኣࡄࡿࡣኣ࠷࡮
࡜ຝᯕⓏ࡚࠵ࡾࡆ࡛࠾ࡼࠉ௑ᚃ஢᝷ࡈࡿࡾ㯔Ⳟࡡࢬࣜࣆ࢒ࢠࢹ࣭࡛ࣛࡊ࡙ࡡ㧏ᗐฺ⏕࡞࠽࠷࡙ࡵᚪ㡪ࡡࢵ࣭ࣜ࡞࡝ࡾ࡛⩻࠻࡙࠷ࡾࠊ
ᮇ◂✪ࡢ⏍◂ࢬࣤࢰ࣭ᇱ♇◂✪᥆㐅஥ᴏࠔࢰࣤࣂࢠ㈻ᕝሔ࡛ࡊ࡙ࡡ⣊≟Ⳟࡡ㧏ᗐฺ⏕࡞㛭ࡌࡾᇱ┑Ⓩ◂✪ࠕࡡୌ⎌࡛ࡊ࡙⾔ࢂࡿࡒࠊ
A novel vector system for rapid preparation of Aspergillus oryzae expression plasmids: its application for
production of Fugu proteins
Manabu Arioka (Department of Biotechnology, The University of Tokyo)

13

S-2
㯔Ⳟ࡞ࡻࡾࣃࢹࣛࢯࢲ࣭࣑⏍⏐࡞ཀྵ࡯ࡌࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪࡡຝᯕ ୷ᒜ₮ୌ㸝᮶ኬ㝌㎨⏍⛁࣬ᚺ⏍ᕝ㸞㯔Ⳟ A. oryzae ࡢࢰࣤࣂࢠ㈻㧏ฦἢ⬗ࢅ᭯ࡊࠉࡱࡒᏭධᛮ࠿ヾࡴࡼࡿ࡙࠷ࡾࡆ࡛࠾ࡼࠉ␏⛸ࢰࣤࣂࢠ㈻⏍⏐ࡡᐙ୹࡛ࡊ࡙Ἰ┘ࡈࡿ࡙࠷ࡾࠊࡆࡿࡱ࡚ࡡ A. oryzae ࢅ⏕࠷ࡒ⏍⏐ࡡモࡲ࡚ࡢࠉᙁງ࡝㧏Ⓠ⌟ࣈ࣓࣭ࣞࢰ࣭ࡡ
㛜Ⓠࠉࢤࢺࣤࡡ᭩㐲໩࡝࡜࡚ᠺᯕ࠿ᚋࡼࡿ࡙࠷ࡾࠊୌ᪁࡚ࠉ⣊≟Ⳟ⏜ᮮࡡࡵࡡ࡛Ẓ࡬࡙㧏➴⏍∸⏜ᮮࡡࢰࣤ
ࣂࢠ㈻ࡡ⏍⏐㔖࠿఩࠷࡞ࡵ࠾࠾ࢂࡼࡍࠉࡐࡡฦよ࡞㛭୙ࡌࡾ࡛⩻࠻ࡼࡿࡾࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪࡡሒ࿈ࡢ
࡮࡛ࢆ࡜࡝࠷ࠊⁿ⩽ࡼࡢࠉ␏⛸ࢰࣤࣂࢠ㈻ࡡ࣓ࢸ࡛ࣜࡊ࡙ࣃࢹࣛࢯࢲ࣭࣑ࡡ⏍⏐ࢅ A. oryzae ࢅ⏕࠷࡙⾔࠷ࠉ㸪⛸ࡡࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄࡡ◒ቪ࡞ࡻࡾຝᯕࢅ᳠ゞࡊࡒࠊ㸦ࠉA. oryzae ࢅ⏕࠷ࡒࣃࢹࣛࢯࢲ࣭࣑ࡡ⏍⏐ 1)ࣃࢹࣛࢯࢲ࣭࣑ࢅⓆ⌟ࡌࡾࡒࡴࡡࣈࣚࢪ
࣐ࢺࡢࠉMultiSite GatewayTM ࢨࢪࢷ࣑࡞ࡻࡽ
α ࢓ ࣐ࣚ ࣭ࢭ &P^ ' 㸧 ࣃࢹ ࣛ ࢯࢲ ࣭࣑ష⿿ࡊࡒࠊࡱࡒ࢞ࣔࣛ࢓࣭࡞ࡢࠉA. oryzae ࠿᭩ࡵኬ㔖࡞ฦἢࡌࡾࢰࣤࣂࢠ㈻࡚࠵ࡾ a-࢓࣐
0 7 
0 7  ࣭ࣚࢭ㸝AmyB㸞ࢅ⏕࠷ࡒࠊamyB ࣈ࣓࣭ࣞࢰ
࣭࠽ࡻࡦ ORF ࡡୖὮ࡞ࣃࢹࣛࢯࢲ࣭࣑㐿ఎᏄࢅࢰࣤࢸ࣑࡞㸧ࢤࣅ࣭㏻⤎ࡊࡒ⼝ྙ㐿ఎᏄᅒ㸦㸣 ࣃ ࢹࣛ ࢯࢲ ࣭࣑ ⏍ ⏐ࡡ ࡒࡴ ࡡ⼝ ྙ ࢰࣤ ࣂࢠ ㈻ࠊ ྘㏻⤎㒂ࢅష⿿ࡊࠉࡐࡿࡑࡿࡡ㏻⤎㒂న࡞ࡢ Kex2 วฦ࡞᣼ථࡊࡒ㒼า 07 ࡢ0H]ᵕࣈࣞࢷ࢓࣭ࢭ࡞ࡻࡽว᩷ࡈࡿࡾࡡ
࡚ࠉ㐗㞫ࡊࡒࣃࢹࣛࢯࢲ࣭࣑࠿ᇰᆀ୯࡞᳠ฝࡈࡿࡾࠊ
᩷㒼าࢅ᣼ථࡊࡒ㸝ᅒ㸦㸞ࠊᐙ୹࡞ࡢࠉᠻࠍࡡ
ࢡ࣭ࣜࣈ࡚⫩⛸ࡊࡒ㸩㔔ᰜ㣬さịᛮ NSAR1
ᰬ(niaD- sC- ǼargB adeA-)2,3)ࢅ౐⏕ࡊࡒࠊࡆࡡᰬࢅ⏕࠷ࡾ࡛ࠉ␏⛸ࢰࣤࣂࢠ㈻ࢅⓆ⌟ࡌࡾ௧አ࡞ࠉptrA ࣏࣭࣭࢜ࡵྱࡴ࡙᭩ኬ㸩ࢪࢷࢴࣈࡡ㐿ఎᏄ᧧ష࠿ྊ⬗࡚࠵ࡾࠊࣃࢹࣛࢯࢲ࣭࣑Ⓠ⌟ࣈࣚࢪ࣐ࢺࢅࠉAspergillus
nidulans sC ࣏࣭࣭࢜࡞ࡻࡽ NSAR1 ᰬ࡞ᙟ㈻㌹ᥦࡊࡒࠊཱིᚋࡊࡒᰬࡡᇰ㣬୕ΰ࡞ᑊࡊ࡙࢙ࢗࢪࢰࣤよᯊࢅ⾔ࡖࡒ⤎ᯕࠉ஢᝷ࡈࡿࡒฦᏄ㔖ࡡࣃࢹࣛࢯࢲ࣭࣑࠿᳠ฝࡈࡿࡒࠊ⮫㐲⏍⏐᮪௲ࢅ᳠ゞࡊࡒ⤎ᯕࠉ5DPY ᾦమᇰ
ᆀ(pH 8.0)࡚᭩ኬ 15.6 mg/L ࡡ⏍⏐㔖࠿ᚋࡼࡿࡒࠊ
㸧ࠉ㸪⛸ࡡࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪ࡞ࡻࡾ ࣃࢹࣛ ࢯࢲ࣭࣑⏍⏐ࡡ᳠ゞḗ࡞ࠉ␏⛸ࢰࣤࣂࢠ㈻ࡡฦよ࡞㛭୙ࡌࡾྊ⬗ᛮࡡ࠵ࡾࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ࡞ࡗ࠷࡙ࠉ⣌⤣Ⓩ࡞◒ቪᰬࢅష⿿ࡊࡒ 1)ࠊⳞమአ㓗ᛮࣈࣞࢷ࢓࣭ࢭ㸝pepA㸞ࠉⳞమහ㓗ᛮࣈࣞࢷ࢓࣭ࢭ㸝pepE㸞ࠉⳞమአ࢓ࣜ࢜ࣛࣈࣞࢷ࢓࣭ࢰ࣭ ࢣࢴ ࢹ㐿 ఎ Ꮔ ࣏࣭ ࣭࢜ ࢰ࣭ࢣࢴࢹ㐿ఎ Ꮔ 

5(7࣏࣭࣭࢜ 

5(7

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’ഁ୕Ὦ 㡷 ᇡ ࣏࣭࣭࢜ 

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5(7

’ഁ୕Ὦ 㡷 ᇡ ࣏࣭ ࣭࢜ ’ഁୖ Ὦ 㡷ᇡ 㐿ఎ Ꮔ◒ ቪ ⏕᩷ ∞ ᅒ㸧2XOWL8LWH,FWH\F^ 92 ࢨࢪࢷ ࣑㸝 ᕞ㸞 ࠽ ࡻࡦIXVLRQ5(7Ἢ㸝ྎ㸞 ࡞ࡻ ࡾ㐿 ఎ Ꮔ◒ ቪ⏕ ᩷∞ ࡡ ష⿿ 14ࢭ㸝alpA㸞㸞
ࠉࢹࣛ࣋ࣈࢲࢩࣜ࣋ࣈࢲࢱ࣭ࢭ㸝tppA㸞
ࠉ࢜ࣜࣂ࢕ࣤᵕࣈࣞࢷ࢓࣭ࢭ㸝palB㸞ࡡ㸪⛸ࡡ㐿ఎᏄࢅ㐽
ࡦࠉMultiSite GatewayTM ࢨࢪࢷ࣑ࡱࡒࡢ fusion PCR Ἢࢅ⏕࠷࡙◒ቪ⏕ DNA ᩷∞䜘ష⿿䛝䛥䟺ᅒ䠄䟻䚯䛙䜒䜏䛴᩷∞䜘 adeA ࣏࣭࣭࢜࡞ࡻࡖ࡙ࣃࢹࣛࢯࢲ࣭࣑⏍⏐ᰬ࡞ᙟ㈻㌹ᥦࡊࠉ┘Ⓩࡡ◒ቪᰬࢅཱིᚋࡊࡒࠊ྘ࣈࣞࢷ࢓࣭ࢭ
㐿ఎᏄ◒ቪᰬࡡࣃࢹࣛࢯࢲ࣭࣑⏍⏐ᛮࢅ 5DPY ᾦమᇰᆀ㸝pH 8.0㸞࡞࠽࠷࡙Ẓ㍉ࡊࡒࠊࡐࡡ⤎ᯕࠉtppAࠉpalBࠉ
pepEࠉalpAࠉpepA ࡡ㡨࡞㐿ఎᏄ◒ቪࡡຝᯕ࠿ヾࡴࡼࡿࠉ࡝࠾࡚ࡵ tppA 㐿ఎᏄ◒ቪᰬࡡࣃࢹࣛࢯࢲ࣭࣑⏍⏐㔖
ࡢ 21.2 mg/L ࡞㐡ࡊࡒࠊ㸨㸞ࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ㸧㔔◒ቪ࡞ࡻࡾࣃ ࢹࣛࢯ ࢲ࣭࣑⏍⏐㔖ࡡቌຊࡈࡼ࡞ࠉࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄࢅ㸧㔔◒ቪࡌࡾࡆ࡛࡞ࡻࡽࠉࣃࢹࣛࢯࢲ࣭࣑⏍⏐㔖ࡡቌຊࢅモࡲࡒ 4)ࠊpalB
࠽ࡻࡦ pepE ྘㐿ఎᏄ◒ቪᰬ࡞ᑊࡊ࡙ࠉargB ࢅ㐽ᢝ࣏࣭࣭࡛࢜ࡊ࡙⏕࠷ tppA 㐿ఎᏄࢅ◒ቪࡊࡒࠊࡆࡿࡼࡡᰬ
ࢅ⏕࠷࡙ࣃࢹࣛࢯࢲ࣭࣑⏍⏐ࢅ⾔ࡖࡒ࡛ࡆࢀࠉtppA, pepE 㐿ఎᏄ㸧㔔◒ቪᰬ࠿᭩ࡵ㧏࠷⏍⏐㔖㸝25.4 mg/L㸞ࢅ♟ࡊࡒࠊࡆࡿࡼࡡ⤎ᯕ࡞ࡻࡽࠉA. oryzae ࢅ⏕࠷ࡒࣃࢹࣛࢯࢲ࣭࣑⏍⏐࡞࠽࠷࡙ࠉࣈࣞࢷ࢓࣭ࢭ㐿ఎ
㔕⏍ᆵ

Ꮔࡡ㸧㔔◒ቪࢅ⾔࠹ࡆ࡛࡚⣑ 1.6 ಶࡡ⏍⏐㔖ቌຊ࡞ᠺຉࡊࡒ㸝ᅒ㸨㸞ࠊᮇ◂✪ࡢࠉA. oryzae ࡞࠽࠷࡙⣌⤣Ⓩ࡝ࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪ࡞ࡻ Ǽ W S S & ࡽ㧏➴⏍∸⏜ᮮࢰࣤࣂࢠ㈻ࡡ⏍⏐㔖࠿୕᪴ࡊࡒิࡴ࡙ࡡౚ࡚࠵ࡾࠊࡱࡒࠉ␏⛸ࢰࣤࣂࢠ㈻ࢅ ǼWSS& Ⓠ⌟ࠉ㸧㔔㐿ఎᏄ◒ቪࢅࡊࡒ࠹࠻࡚ࠉࡐࡿ࡚ࡵ ǼSHS* ࡝࠽㸧ࢪࢷࢴࣈࡡ㐿ఎᏄ᧧ష࠿ྊ⬗࡚࠵ࡾࠊࡈ
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ࡼ࡞ࠉᑚ⬂మࢨࣔ࣋ࣞࣤ࡝࡜ฦἢ㛭㏻㐿ఎᏄࡡᅒ 㸨  ࣈ ࣞ ࢷ ࢓ ࣭ ࢭ 㐿 ఎ Ꮔ ◒ ቪ ࡞ ࡻ ࡾ & RU^̲FH ࢅ ⏕ ࠷ ࡒ᧧షࢅࡌࡾࡆ࡛࡚ࠉA. oryzae ࠿᭯ࡌࡾࢰࣤࣂࢠࣃࢹ ࣛࢯ ࢲ࣭ ࣑ ⏍⏐ 㔖ࡡ ቌຊ ㈻㧏⏍⏐⬗ࡡ࣎ࢷࣤࢨࣔࣜ࠿ᝨࡊࡲ࡝ࡂⓆᥱ
ࡈࡿࡾࡆ࡛࠿௑ᚃ᭿ᙽࡈࡿࡾࠊ
ᮇ◂✪ࡢ⏍◂ࢬࣤࢰ࣭ᇱ♇◂✪᥆㐅஥ᴏࠔࢰࣤࣂࢠ㈻ᕝሔ࡛ࡊ࡙ࡡ⣊≟Ⳟࡡ㧏ᗐฺ⏕࡞㛭ࡌࡾᇱ┑Ⓩ◂✪ࠕࡡୌ⎌࡛ࡊ࡙⾔ࢂࡿࡒࠊ1) 㔘ࡼࠉ➠㸩ᅂ⣊≟ⳞฦᏄ⏍∸Ꮥࢤࣤࣆ࢒ࣝࣤࢪさ᪠㞗ࠉp. 39
2) Jin et al., FEMS Microbiol. Lett., 239, 79-85 (2004)3) ໪ᮇࡼࠉ⏍∸ᕝᏕఌヽࠉ83, 277-279 (2005)4) 㔘ࡼࠉ᪝ᮇ㎨ⰹ໩Ꮥఌ 2005 ᖳᗐኬఌさ᪠㞗ࠉp. 240

Effects of protease gene disruption on human lysozyme production by Aspergillus oryzae
Jun-ichi Maruyama(Dept. of Biotechnology, Univ. of Tokyo)

15

S-3
麹菌細胞壁のリモデリング：細胞内に生産させたタンパク質を培養上清へ回収するための戦略
堀内裕之（東京大学大学院農学生命科学研究科）

高等動植物から微生物までに及ぶ近年の急速なゲノムプロジェクトの進行は、人類の生活をより良くする
ための新たな可能性を示す一方で膨大な数の機能不明なタンパク質の存在を明らかにしてきた。そこでそれらタンパク質を調製しその機能を解析する手法を確立することは、ゲノムデータの高度利用において必要不可欠のことである。本研究はそのような状況の下で菌体内タンパク質の簡便な調製法として、糸状菌の細胞壁を人為的に改変することにより菌体内のタンパク質を培養上清へ漏出させ回収する手法の開発を目的としたものである。
麹菌 Aspergillus oryzae は古来より日本において清酒、醤油、味噌等の醸造、製造に利用されてきており、長い歳月にわたって人間の生活に密接にかかわってきたことからその安全性が認められており、その全ゲノム DNA 配列も決定されており、発現を制御可能な強力なプロモーター等も利用できることから、タンパク質生産の場としての利用に非常に適している。麹菌の細胞壁の主要構成成分は N-アセチルグルコサミンが β−1,4結合でつながったポリマーであるキチンとグルコースが β−1,3 結合でつながった β-1,3-グルカンからなる。
そこでこれらの分解酵素遺伝子の発現を人為的に制御することにより細胞質タンパク質の細胞外への漏出への効果を検討した。
麹菌キチナーゼ遺伝子の単離とその高発現
Aspergillus 属の糸状菌においてはすでに Aspergillus nidulans、Aspergillus fumigatus の全ゲノム配列が公開されているが、これらの配列情報から、Aspergillus 属糸状菌には十数種のキチナーゼ遺伝子が存在することが推定されており、A. oryzae にも同程度の数のキチナーゼ遺伝子の存在が推定された。そこでそれらの中から
A. oryzae の細胞壁中のキチンを効率良く分解できるキチナーゼをコードする遺伝子を選択するため、我々の研究室において機能解析を行っている A. nidulans の 18 種のキチナーゼ遺伝子の中から、細胞壁キチンの分解に適したキチナーゼをコードすると考えられる chiA、chiB に着目した。A. nidulans において chiB の発現は培養後期に誘導され、その自己溶菌において重要な役割を果たすことが示されており(山崎ら、投稿準備中)、chiAは高発現させることによりある種の条件下で菌糸の溶菌に効果があることが示唆されていた。chiA、chiB の
A. oryzae のオーソログ（ここでは A. nidulans の遺伝子との区別のために AochiA、AochiB と呼ぶ）に対し、A.
oryzae のゲノム情報を利用してそれぞれ遺伝子を単離し、その発現について検討した。その結果、これら遺伝子の発現は、A. nidulans の chiA、chiB の発現様式とはパターンを異にしており、通常の培養では A. oryzae
では AochiA の発現量が多く、AochiB の発現はほとんど見られなかった。AochiA、AochiB をそれぞれ A. oryzaeの amyB プロモーターの下流につないだ形で A. oryzae の thiA locus に導入し、amyB プロモーターの高発現条件である培地の炭素源をマルトースにしたプレート上で培養した場合の影響について検討したが、これらの遺伝子を単独で高発現させただけでは菌糸の形態に変化は見られなかった。
グルカナーゼ遺伝子の単離とその高発現の効果
次に β−1,3-グルカナーゼ遺伝子を単離してその高発現の効果を検討した。A. oryzae のゲノム上にはキチナーゼの場合と同様 β-1,3-グルカナーゼと相同性を有する遺伝子が多数存在することが推定されたため自己溶菌と比較的類似した環境であると考えられる炭素源を除いた状況での A. oryzae の EST 情報より 3 種のグルカ
ナーゼをコードする遺伝子を同定し、その全長を単離した。これらの遺伝子を gluA、gluB、gluC と命名し、それぞれ amyB プロモーター下、sC locus で高発現できる株を作製した。その結果、gluA、gluB を高発現できる株では培地の炭素源をマルトースにしたプレート上では、非常に低頻度ではあるが菌糸の溶菌が観察された。一方、A. oryzae の菌体に対し、外部から作用させることで細胞壁の溶解に効果があることが示唆されて）いる Bacillus circulans の分泌性 β−1,3-グルカナーゼをコードする bglH 遺伝子 1, 2 についてもその成熟タンパ

16

ク質をコードする部分を amyB 遺伝子の分泌シグナルをコードする部分に in frame でつないだ amyB-bglH キ
メラ遺伝子を amyB プロモーター下で高発現できる株を作製したところ amyB プロモーターの発現を誘導できる条件下ではやはり非常に低頻度ではあるが菌糸の溶菌が観察された。
キチナーゼ遺伝子とグルカナーゼ遺伝子をともに高発現できる株の作製
キチナーゼ遺伝子、グルカナーゼ遺伝子、それぞれ単独では菌体の溶菌に対して効果が小さいことが明らかになったため、次にキチナーゼ遺伝子とグルカナーゼ遺伝子を同時に amyB プロモーター下で高発現できる株を２種のキチナーゼ遺伝子と４種のグルカナーゼ遺伝子の全ての組み合わせで作製した。その結果、AochiB と各種グルカナーゼ遺伝子を導入し高発現させた株では、グルカナーゼ遺伝子のみを高発現させた場合と比較して差が見られなかったのに対し、AochiA と gluA、AochiA と gluB、AochiA と amyB-bglH 遺伝子を同時に導入した株では、炭素源をマルトースとしたプレート上で頻繁に菌糸の溶菌が見られた。さらに AochiAと gluB を amyB プロモーターにつないだ株について、これら遺伝子の発現を抑える液体培養条件で 2〜３日間培養した後、発現を誘導する液体培養条件にシフトして経時的に菌体重量を測定したところ、AochiA と gluBの発現を誘導した株は同様の条件で培養した野生株に対して菌体重量が 3-4 割程度少ないことが明らかになった。さらにこれら培養菌体を顕微鏡で観察したところ、内容物が漏出したような菌糸が見られた。これらのことから、AochiA と gluB の同時高発現によりプレート上でも、液体培養においても菌糸の溶菌が誘導できることが明らかになった。そこで現在、細胞質タンパク質の挙動について検討を行っている。
以上の結果から、ある種のキチナーゼとグルカナーゼを同時に高生産させることにより A. oryzae の菌糸を溶菌させることが可能であることが明らかになった。Aspergillus 属糸状菌の菌糸には隔壁が存在するが、隔壁の中央に穴があるため細胞質はつながっている。しかし菌糸細胞壁に何かの原因で損傷が生じ細胞質のタンパク質が細胞外へ漏出するような状況になった場合、Woronin body と呼ばれるオルガネラが隔壁の穴を塞ぐことにより、損傷の影響が隔壁を越えて広がらないようにしている。A. oryzae においては近年 Woronin）body を構成するタンパク質をコードする遺伝子 Aohex1 が単離されその機能解析が行われている 3 。先に述べたキチナーゼ遺伝子、グルカナーゼ遺伝子を同時に高発現できる株において Aohex1 を破壊すれば、細胞質のタンパク質等を細胞外へより効率的に漏出させることが期待できることから、今後は、最近開発された４種の栄養要求性を持つ A. oryzae の株 4)を親株としてこれらを組み合わせた株を作製しその効果を検討してゆきたい。
本研究は生研センター基礎研究推進事業「タンパク質工場としての糸状菌の高度利用に関する基盤的研究」の一環として行われた。
1) Horikoshi, K. and Iida S. Nature 183: 186-187 (1959)
2) Yamamoto, M., Ezure, T., Watanabe, T., Tanaka, H. and Aono, R. FEBS Lett. 433: 41-43 (1998)
3) Maruyama, J., Juvvadi, P. R., Ishi, K. and Kitamoto, K. Biochem. Biophys. Res. Commun. 331: 1081-1088 (2005)
4) Jin, F. J., Maruyama, J., Juvvadi, P. R., Arioka, M. and Kitamoto, K. FEMS Microbiol. Lett. 239: 79-85 (2004)
Strategy to promote the leakage of intracellular proteins into culture supernatants by altering the cell wall
structure of Aspergillus oryzae
Hiroyuki Horiuchi(Department of Biotechnology, The University of Tokyo)

Ở17

7FWLR

S-4
㯑Ⳟࡡᾦమᇰ㣬࡚ᅖమᇰ㣬࡛ྜྷᵕࡡ㐿ఎᏄⓆ⌟ࢅᐁ⌟ࡌࡾࡒࡴࡡᡋ␆
ᆊᮇ࿰ಆ㸝㒿㢦⥪ྙ◂✪ᡜ㸞㯑Ⳟ㸝Aspergillus oryzae㸞ࡢࡐࡡ⡷㯑࡝࡜ࡡᅖమᇰ㣬࡞࠽ࡄࡾ㧏࠷ࢰࣤࣂࢠ㈻⏍⏐࣬ฦἢ⬗ງ࠾ࡼࠉ㔂㏸⏐ᴏࠉ㓕⣪⏍⏐࡝࡜ࡡฦ㔕࡚ཿࡂ࠾ࡼᗀࡂฺ⏕ࡈࡿ࡙࠷ࡾࠊࡆࡡ≁ᚡࡢࠉ㯑Ⳟ࠿ᅖమᇰ㣬࡛࠷࠹⎌ሾ࡞ᚺ➽ࡊࠉฦἢ࡞㛭୙ࡌࡾࢰࣤࣂࢠ㈻ࡷ㓕⣪࡝࡜ࡡ㐿ఎᏄⓆ⌟
ࢅโᚒࡊ࡙࠷ࡾࡒࡴ࡛⩻࠻ࡼࡿࡾ࠿ࠉࡐࡡฦᏄ⏍∸ᏕⓏ⫴ᬊࡢࠉ࠷ࡱࡓレ⣵࡞ࡢよ᪺ࡈࡿ࡙࠷࡝࠷ࠊᅖమᇰ㣬≁␏Ⓩ࡝㐿ఎᏄⓆ⌟โᚒࡢ glaB 㐿ఎᏄࡡ◂✪࡞ጙࡱࡽࠉࡆࡿࡱ࡚࡞ melBࠉpepAࠉ
AOS 㐿ఎᏄ⩄࡝࡜㟸ᖏ࡞ኣᩐࡡᅖమᇰ㣬≁␏Ⓩ㐿ఎᏄ࠿Ꮛᅹࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ࡙ࡀ
ࡒࠊࡆࡡࡆ࡛࠾ࡼࠉ㯑Ⳟࡡᅖమᇰ㣬୕࡚ࡡᵕࠍ࡝ᙟ㈻ࡢࠉᅖమᇰ㣬≁␏Ⓩ࡝㐿ఎᏄⓆ⌟โᚒ࡞౪Ꮛࡊ࡙࠷ࡾ࡛஢᝷ࡈࡿࡾࠊ㏣࡞ࠉᅖమᇰ㣬࡚ࡡ㐿ఎᏄⓆ⌟ࢅᾦమᇰ㣬୯࡚ᐁ⌟ࡌࡾࡆ࡛࠿࡚ࡀࡿࡣᅖమᇰ㣬୕࡚ࡡᙟ㈻ࢅᾦమᇰ㣬࡚ࡵ්⌟ࡈࡎࡼࡿࡾࡆ࡛࠿᭿ᙽࡈࡿࡾࠊ
‫ې‬ᅖమᇰ㣬ᚃ᭿ࡡ㐿ఎᏄⓆ⌟ࢅโᚒࡌࡾ㌹෕โᚒᅄᏄ ATFBᅖమᇰ㣬࡞࠽࠷࡙ᾦమᇰ㣬࡛ྜྷ➴ࡡ㐿ఎᏄⓆ⌟ࢅᐁ⌟ࡈࡎࡾࡒࡴ࡞ࡢࠉᅖమᇰ㣬ࢅវ▩ࡌࡾࣝࢬࣈࢰ࣭࠾ࡼ㌹෕โᚒᅄᏄࡡ㛣ࡡࢨࢡࢻࣜఎ㐡⣌ࢅᨭንࡌࡾࡆ࡛࡚ᐁ⌟ྊ⬗࡛⩻࠻ࡼࡿࡾࠊ㌹෕โᚒᅄᏄࢅᨭንࡌࡾࡆ࡛࡚㐿ఎᏄⓆ⌟ࢅ├᥃Ⓩ࡞ን໩ࡈࡎࡼࡿࡾྊ⬗ᛮ࠿㧏࠷࠿ࠉ㌹෕โᚒᅄᏄࢅ༟⣟࡞㧏Ⓠ⌟ࡊࡒࡓࡄ࡚ࡢࠉࡐࡿ࡞โᚒࡈࡿࡾ㐿ఎᏄ࠿Ὡᛮ໩ࡌࡾ
ࡆ࡛ࡢᑛ࡝࠷ࡻ࠹࡚࠵ࡾࠊ࡛ࡆࢀ࠿ࠉฦ⏍Ꮔᙟᠺࡡ࣏ࢪࢰ࣭࣭ࣝ࢟ࣖࣝࢰ࣭࡚࠵ࡾ brlA 㐿ఎᏄࡢ㧏Ⓠ⌟ࡈࡎࡾࡓࡄ࡚ୖὮ㐿ఎᏄࡡ㌹෕࠿Ὡᛮ໩ࡊࠉฦ⏍Ꮔᙟᠺ࡛ࡨࡌࡱ࠷࠹々㞟࡝Ⓠ⏍㐛⛤ࢅᏰ஡ࡈࡎࡾࡆ
ᅖమᇰ㣬࡛࠿࡚ࡀࡾࠊ
ࡆࡡ brlA ࡡ≁ᚡ࡛ࡊ࡙ࠉ
㌹෕โᚒᅄᏄࡡ㐿ఎᏄ⮤㌗࠿㌹෕ࣝ
࡚࣊ࣜโᚒࡈࡿ࡙࠷ࡾࡆ࡛࠿ᣪࡅࡼ
ࡿࡾࡆ࡛࠾ࡼࠉᠻࠍࡢᅖమᇰ㣬᫤࡞㐿ఎᏄⓆ⌟࠿ንິࡌࡾ㌹෕โᚒᅄᏄࢅぜࡗࡄࠉࡆࡿࢅ㧏Ⓠ⌟ࡈࡎࡾࡆ࡛
࡛ࡊࡒࠊࡱࡍ㯑Ⳟ EST ࢸ࣭ࢰ࣭࣊ࢪ
ᾦమᇰ㣬࡞࠽࠷࡙ᅖమᇰ㣬᮪௲࡚ฝ⌟㢎ᗐࡡ(̲)R]K㧏࠷㌹෕โᚒᅄᏄ㐿ఎᏄࢅࣛࢪࢹ࢓
ࢴࣈࡊࠉḗ࡞ࡐࡿࡼࡡ㐿ఎᏄⓆ⌟ࣂK ᇰ㣬᫤㛣ࢰ࣭ࣤࢅㄢ࡬ࡒࠊࡐࡡ⤎ᯕࠉᅖమᇰ
㣬᮪௲࡚Ⓠ⌟࠿ንິࡌࡾ bZIP ࢰ࢕
Fig1 ATFB ࡞โᚒࡈࡿࡾ㐿ఎᏄ⩄ࡡࡨࡌࡱᅖమᇰ㣬୯ࣈ㌹෕โᚒᅄᏄ㐿ఎᏄࢅぜ࠷ࡓࡊࠉ࡚ࡡⓆ⌟ࣂࢰ࣭ࣤࠊATFB ࡢᇰ㣬ᚃ᭿࡞࠽࠷࡙ྜྷㄢࡊ࡙
ࡆࡿࢅ atfB ࡛ྞ௛ࡄ࡙㐿ఎᏄ㧏Ⓠ⌟Ⓠ⌟୕᪴ࡌࡾ㐿ఎᏄ⩄ࢅโᚒࡊ࡙࠷ࡾࠊ
ᰬࢅషᠺࡊࡒࠊ࣏ ࢕ ࢠ ࣞ࢓ ࣝ ࢕ࢅ ฺ ⏕ࡊ࡙ ࡆࡡ
atfB 㧏Ⓠ⌟ᰬࢅよᯊࡊࡒ⤎ᯕࠉᅖమᇰ㣬≁␏ⓏⓆ⌟ࢅ♟ࡌ࢜ࢰ࣭ࣚࢭ㐿ఎᏄ catA ࢅྱࡳ 30
㐿ఎᏄࡡⓆ⌟ࢅ୕᪴ࡈࡎࡾࡆ࡛࡞ᠺຉࡊࡒࠊࡆࡿࡼⓆ⌟࠿୕᪴ࡊࡒ㐿ఎᏄ⩄ࡡࡨࡌࡱᅖమ
ᇰ㣬୯࡚ࡡⓆ⌟ࣂࢰ࣭ࣤࢅㄢ࡬ࡒ⤎ᯕࠉᇰ㣬ᚃ᭿࡞࠽࠷࡙ࡆࡿࡼ㐿ఎᏄࡡⓆ⌟࠿ྜྷㄢࡊ࡙
㧏ࡂ࡝ࡖ࡙࠷ࡒࠊࡗࡱࡽࠉࡆࡿࡼࡢᅖమᇰ㣬≁␏ⓏⓆ⌟ࢅ♟ࡌ㐿ఎᏄ࡚࠵ࡽࠉᠻࠍࡢࡆࡡ
ᅖమᇰ㣬≁␏Ⓩ㐿ఎᏄⓆ⌟ࢅᾦమᇰ㣬୯්࡚⌟ࡌࡾࡆ࡛࡞ᠺຉࡊࡒࠊ

18‫ࡾࡻ࡞࢕ࣝ࢓ࣞࢠ࢕࣏ې‬ᅖమᇰ㣬࡚ࡡ㐿ఎᏄⓆ⌟ࣂࢰ࣭ࣤࡡよᯊ ⡷㯑ࠉ㓲ἔ㯑ࠉࡨࡌࡱᇰ㣬ࡢ⏐ᴏⓏ࡞ࡨࡌࡱ
+LJ㺤㺇㺍㺰㺆㺯㺇࡞ࡻ ᅖమ
㔔さ࡝ᅖమᇰ㣬ࡡ᪁Ἢ࡚࠵ࡾࡆ࡛࠾ࡼࠉ
ࡾ྘ᅖమᇰ㣬࡞࠽ ᇰ㣬ࢨࡆࡿࡼࡡᇰ㣬㐛⛤࡞࠽ࡄࡾ㐿ఎᏄⓆ⌟ࢡࡄࡾ㐿ఎᏄⓆ⌟ࣂࢻࡡ⤊᫤ን໩ࢅ࣏࢕ࢠࣞ࢓ࣝ࢕࡞࡙ふᐳࢰ࣭ࣤࡡよᯊ
ࣜࡊࡒࠊࡐࡡ⤎ᯕࠉ⡷㯑ཀྵࡦࡨࡌࡱᇰ㣬࡚Ẓ⋙ࡢࠉᇰ㣬୯᭿࠾ࡼᚃ᭿࡫ࡡ⛛⾔㐛⛤࡚㐿Ќን᭜Ⅴᑊ↯ఎᏄⓆ⌟ࣂࢰ࣭ࣤࡡን᭜Ⅴ࠿࠵ࡽࠉ㯑Ⳟ᮪௲   
⣵⬂හ࡚ࢱ࢕ࢻ࣐ࢴࢠ࡝ን໩࠿㉫ࡆࡖ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡨࡌࡱᇰ㣬࡞⡷㯔㓲ἔ㯔࠽࠷࡙ࡢࠉࡆࡡን᭜Ⅴ࠿Ẵ୯Ⳟ⣊ᙟᠺ᭿࡞ルᙔࡌࡾࡆ࡛࠾ࡼࠉࡆࡡ㐿ఎᏄⓆ⌟ࡡንິࡢẴ୯Ⳟ⣊࠾ࡼฦ⏍Ꮔᙟᠺ࡛࠷࠹
ᙟឺን໩ࢅ⾪ࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾࠊ ௑ᅂࡡᠻࠍࡡよᯊ࡞ࡻࡽࠉን᭜ⅤᚃࡡЌን᭜Ⅴ㐿ఎᏄⓆ⌟ࡡୌ㒂ࡢ atfB ࢅ㧏Ⓠ⌟ࡈࡎ 
ࡾࡆ࡛࡚ᾦమᇰ㣬࡞࠽࠷්࡙⌟࡚ࡀࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊ௙ࡡⓆ⌟ࣂࢰ࣭ࣤࢅ♟ࡌᅖమᇰ㣬≁␏Ⓩ㐿ఎᏄⓆ⌟ࡵࠉ௑ᚃࡡよᯊ࡞ࡻࡽᾦమᇰ㣬්࡚⌟ࡌࡾࡆ࡛࠿ྊ⬗࡛࡝ࡽࠉᵕࠍ࡝ᅖమᇰ㣬≁␏Ⓩᙟ㈻ࢅᾦమᇰ㣬࡚ᐁ⌟࡚ࡀࡾ࡞㐢࠷࡝࠷ࠊ‫ې‬ㅨ㎙ᮇ◂✪ࡢ⏍◂ࢬࣤࢰ࣭ᇱ♇◂✪᥆㐅஥ᴏࠔࢰࣤࣂࢠ㈻ᕝሔ࡛ࡊ࡙ࡡ⣊≟Ⳟࡡ㧏ᗐฺ⏕࡞㛭
ࡌࡾᇱ┑Ⓩ◂✪ࠕࡡୌ⎌࡛ࡊ࡙⾔ࢂࡿࡒࠊHow to express the solid-state specific genes of Aspergillus oryzae in liquid culture
Kazutoshi Sakamoto
National Research Institute of Brewing

19

8
ࣈࣞࢷ࢛࣭࣑࠾ࡼぜࡒ㯔Ⳟ㸝 &VSHUJLOOXVRU^̲FH 㸞ࡡࢰࣤࣂࢠ㈻⏍⏐ᶭᵋᒷୖ࿰⿩࣬⧂⏛ ೸㸝≺❟⾔ᨳἪெ 㒿㢦⥪ྙ◂✪ᡜ㸞 ࠘ࡢࡋࡴ࡞࠙
㯔Ⳟࡢࠉΰ㒿ࡷ↕㒽ࠉ࿝ძࡷ㓲ἔ࡝࡜ᠻ࠿ᅗࡡఎ⤣Ⓩ㔂㏸⏐ᴏ࡞࠽࠷࡙ࠔ㯔ࠕ࡛ࡊ࡙౐⏕ࡈࡿࡾࠊ㯔Ⳟࡡ᭩ኬࡡ≁ᚡࡢࠉࡐࡡ㦣␏Ⓩ࡝ࢰࣤࣂࢠ㈻⏍⏐ᛮ࡚࠵ࡽࠉኣࡂࡡࢰࣤࣂࢠ㈻ࡢࠉ≁࡞ࠔᅖమᇰ㣬ࠕ࠷ࢂࡹࡾࠔ㯔㏸ࡽࠕࢅ⾔ࡖࡒ࡛ࡀ࡞ኣ㔖࡞⏍⏐ࡈࡿࡾࠊ
┷ᰶ⏍∸ࡡࢰࣤࣂࢠ㈻⏍⏐࡞ࡗ࠷࡙ࡢࠉ㓕ẍࡷࣃࢹࡡ⣵⬂ࢅ⏕࠷࡙ࠉ㌹෕โᚒ࠾ࡼ P73& ࡡࢪࣈࣚ࢕ࢨࣤ
ࢡ࡛ᰶአ࡫ࡡ㍲㏞ࠉࣛ࣍ࢮ࣭࣑࡚ࡡ⩳ゼโᚒࡷᑚ⬂మ࡚ࡡࣆ࢚࣭ࣜࢸ࢔ࣤࢡ࡛⢶㙈ಞ㣥ࠉᑚ⬂㍲㏞ࡡโᚒࡷࢮ࣭ࢷ࢔ࣤࢡࠉ⣵⬂አ࡫ࡡฦἢ࡞⮫ࡾࡱ࡚ࠉฦᏄ࡚ࣝ࣊ࣜレ⣵࡞᳠ゞࡈࡿ࡙࠷ࡾࠊ㯔Ⳟ࡚ࡵ࡮࡯ྜྷᵕࡡ
࣒࢜ࢼࢫ࣑࡞ࡻࡽࢰࣤࣂࢠ㈻⏍⏐࠿ࢤࣤࢹ࣭ࣞࣜࡈࡿ࡙࠷ࡾࡵࡡ࡛⩻࠻ࡼࡿࠉᐁ㝷࡞ኣࡂࡡᐁ㥺ࢸ࣭ࢰ࠿ࡐࡿࢅᙁࡂ♟ြࡊ࡙࠷ࡾࠊࡊ࠾ࡊࠉ㔂㏸⏐ᴏ୕㔔さ࡚᭩ࡵ㔔さ࡝ࠔᅖమᇰ㣬ࠕ࡚ࡡࢰࣤࣂࢠ㈻⏍⏐ᶭᵋ࡞ࡗ࠷࡙ࡢࠉ༎ฦ࡞◂✪ࡈࡿ࡙࠷ࡾ࡛ࡢゕ࠷࠿ࡒ࠷ࠊࡐࡆ࡚ࠉᠻࠍࡡ◂✪ࢡ࣭ࣜࣈ࡚ࡢࠉᅖమᇰ㣬࡞╌┘ࡊࠉ㯔Ⳟ㓕⣪⏍⏐࣒࢜ࢼࢫ࣑ࡡධమാࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡞◂✪ࢅ⾔ࡖ࡙ࡀࡒࠊ
࠘㯔Ⳟ⏍⏐ࢰࣤࣂࢠ㈻ࡡ 5URWHRPHよᯊ࠙ࡆࡿࡱ࡚ࡡ಴ࠍࡡ㓕⣪ᏕⓏ࡝◂✪࠾ࡼࠉᅖమᇰ㣬࡚ࡢᾦమᇰ㣬࡞Ẓ࡬ኣ⛸㢦ࡡ㓕⣪ࡡ⏍⏐㔖࠿୕᪴ࡌࡾ
஥࠿᪺ࡼ࠾࡛࡝ࡖ࡙ࡀ࡙࠷ࡾࠊࡊ࠾ࡊࠉᅖమᇰ㣬࡛ᾦమᇰ㣬࡛ࡡ㓕⣪⏍⏐ࡡᕣ␏࡞ࡗ࠷࡙ࠉ㫵▌Ⓩ࡝❟ሔ࠾ࡼࡡ◂✪ࡢ࡝ࡂࠉ⥪ྙⓏ࡝㆗ㄵࢅࡌࡾࡆ࡛࠿㞬ࡊ࠾ࡖࡒࠊࡐࡆ࡚ࠉ㯔Ⳟ࠿⏍⏐ࡌࡾධࢰࣤࣂࢠ㈻࡞ࡗ࠷
࡙ࠉࣈࣞࢷ࢛࣐ࢠࢪⓏᡥἪ࡞ࡻࡽよᯊࢅ⾔࠷ࠉධమാࢅᢍᥩࡊࡒ୕࡚ࠉ಴ࠍࡡ≁ᚡ࠵ࡾࢰࣤࣂࢠ㈻ࡡ⏍⏐࣒࢜ࢼࢫ࣑࡞ࡗ࠷࡙よᯊࡌࡾࡆ࡛࡛ࡊࡒࠊ
&' ()*+,-./0123456"789:;<=>?
!"#$
%"#$
Group 1 GlaB, DppV et.al.

図１固体培養と液体培養でのタンパク質生産量の比較










Total protein produciton of SSC and SMC
90

AAA AAAAAA AAA80µ g protein/ mg dry mycelia70Group 2 GlaA, XynG2, AgdA, CelB, lactase, rAspf9 et.al.

60

AAAAA AAAAAAA

50
40Group 3 Oryzin, Xyn G1, Xyn F3, catalase et.al.

30

AAAAAA

20

AAAAA A

10
0

SSC

SMC

Group 4 TAA, Bgl et.al.
AAAAA AAAAAAA⥑⨮Ⓩ࡝ࣈࣞࢷ࢛࣭࣑よᯊࡡ⤎ᯕࠉᾦమᇰ㣬࡞Ẓ㍉ࡊ࡙ࠉᅖమᇰ㣬࡚ࡢ㯔Ⳟࡡ⥪ࢰࣤࣂࢠ㈻⏍⏐㔖ࡢ㸪ಶ௧୕࡞୕᪴ࡊ㸝ᅒ㸦㸞ࠉ⏍⏐ࡈࡿࡾࢰࣤࣂࢠ㈻ࡡ⛸㢦ࡵ␏࡝ࡖ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡱࡒࠉ⏍⏐ࡈࡿࡒࢰࣤࣂࢠ㈻ࡢࠉ
ࠔᅖమᇰ㣬࡚ࡻࡽኣࡂ⏍⏐ࡈࡿࡾࡵࡡࠕ㸝ࢡ࣭ࣜࣈ㸦㸯,OF')SS; ࡝࡜㸞ࠉࠔᾦమᇰ㣬
࡚ࡻࡽኣࡂ⏍⏐ࡈࡿࡾࡵࡡ㸝ࢡ࣭ࣜࣈ㸧㸯,OF&=^Q, ࡝࡜㸞ࠕࠉࠔ୦ᇰ㣬᮪௲࡚⏍⏐ࡈࡿࡾࡵࡡ㸝ࢡ࣭ࣜࣈ
㸨㸯=^O, ࡝࡜㸞ࠕࠉࠔᅖమᇰ㣬࡚ࡢᇰᆀ୯࡞⏍⏐ࡈࡿࠉᾦమᇰ㣬࡚ࡢ⣵⬂ባ࡞ኣࡂᑻᅹࡌࡾࡵࡡ㸝ࢡ࣭ࣜࣈ
㸩㸯'JO9&& ࡝࡜㸞ࠕ࡞ฦ㢦ࡈࡿࡒ㸝ᅒ㸧㸞ࠊ྘ࢡ࣭ࣜࣈࢅ௥⾪ࡌࡾࢰࣤࣂࢠ㈻㸝࣏࣭࣭࢜ࢰࣤࣂࢠ㈻㸞
ࡡ㌹෕よᯊ⤎ᯕ࠾ࡼࠉᅖమᇰ㣬࡚ࡢ㌹෕โᚒ࡞ຊ࠻ࠉ㌹෕ᚃࡡโᚒࡵ㔔さ࡚࠵ࡾ஥࠿♟ြࡈࡿࡒࠊ
20

࠘,+5 ⼝ྙ࣏࣭࣭࢜ࢰࣤࣂࢠ㈻࡞ࡻࡾິឺよᯊ࠙྘ࢡ࣭ࣜࣈࡡ࣏࣭࣭࢜ࢰࣤࣂࢠ㈻ࡡິឺ࡞ࡗ࡙ࠉࡈࡼ࡞レ⣵࡝よᯊࢅ㐅ࡴࡾࡒࡴࠉᾦమᇰ㣬࡚ࡵࠉᅖమ
ᇰ㣬࡛ྜྷᵕࡡ㧏Ⓠ⌟࠿ྊ⬗࡝ FP^' ࣈ࣓࣭ࣞࢰ࣭ࢅ౐⏕ࡊ࡙྘⛸࣏࣭࣭࢜ࢰࣤࣂࢠ㈻ࡡິឺ࡞ࡗ࠷࡙よᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉࢡ࣭ࣜࣈ㸦ࡡࢰࣤࣂࢠ㈻ࡢࠉᾦమᇰ㣬୯࡚㧏Ⓠ⌟ࡈࡎ࡙ࡵࠉᇰᆀ୯࡞⏍⏐ࡈࡿࡍ㸝ᇰ㣬㸩㸥᫤㛣㸞ࠉ⣵⬂ࣆࣚࢠࢨࣘࣤ㸝⣵⬂ባࢅྱࡳ㸞࡞᳠ฝࡈࡿࡾࡵࡡ࠿Ꮛᅹࡊࡒ㸝ᅒ㸨㸞ࠊࡱࡒࠉ,OF'*,+5ࡷ )SS;*,+5 ࡡࡻ࠹࡞⣵⬂හ࡞ࣂࢴࢲ≟࡞ᑻᅹࡌࡾࡵࡡࡷࠉ'JO*,+5 ࡷ 9&&*,+5 ࡡࡻ࠹࡞⣵⬂ባཀྵࡦᾦ⬂࡞ᑻᅹࡌࡾࡵࡡ࡝࡜ࠉ࣏࣭࣭࢜ࢰࣤࣂ
ࢠ㈻ࡡ㐢࠷࡞ࡻࡽᑻᅹࣂࢰ࣭ࣤ࠿␏࡝ࡖ࡙

図２ 液体培養でのマーカータンパク質の局在性解析
kD

1

kD

2

1 2

kD

࠷ࡒࠊࡆࡡࡆ࡛࠾ࡼࠉ㌹෕ᚃโᚒ࡞࠽࠷࡙ࢰ
ࣤࣂࢠ㈻⏍⏐ࡡ୕࡚ࡡ࣍ࢹࣜࢾࢴࢠ࠿々ᩐᏋᅹࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡱࡒࠉ㯔Ⳟ࡚ࡢࠉ⣵⬂ባ࠿ࢰࣤࣂࢠ㈻⏍⏐ࡡ୕࡚㝸ባ࡞࡝ࡽ

150
100

*

75
50
37

⏐ࡈࡿࡾࢰࣤࣂࢠ㈻࡞ࡗ࠷࡙ࡢࠉ⣵⬂හࡡ⺧

25
20කᙁᗐ࠿ኬᖕ࡞΅ᑛࡊࡒࡆ࡛࠾ࡼࠉ⣵⬂හ࡞

200

200

࠹ࡾ஥࠿♟ࡈࡿࡒࠊ࡝࠽ࠉᾦమᇰ㣬࡚ኣࡂ⏍

kD

1 2

150
100
75

*

DppV

*

100

37

37

25
20

25
15

25
15

15

10

10

100

50

50

37

37

TAA

*

150
75

75

50

GlaB

*

150

75

1 2

200

200

150
100

50

10

kD

1 2

25
20
10

BglA

GlaA

ೳ⏻ࡌࡾࡆ࡛࡝ࡂⳞమአ࡞ฦἢࡈࡿ࡙࠷ࡾ
ࡵࡡ࡛ᛦࢂࡿࡒࠊ

1.Mycelial fraction (including cell-wall) ２．
Extracellular fraction࠘ࡱ࡛ࡴ࠙
ࡆࡿࡱ࡚ࡡよᯊࡡ⤎ᯕ࠾ࡼࠉᅖమᇰ㣬࡛ᾦమᇰ㣬࡚ࡢ⏍⏐ࡈࡿࡾࢰࣤࣂࢠ㈻ࡡ㔖ཀྵࡦ⛸㢦࠿ኬࡀࡂ␏࡝
ࡾࡆ࡛࠿᪺▉࡞♟ࡈࡿࡒࠊࡱࡒࠉᅖమᇰ㣬࡚ࡡࢰࣤࣂࢠ㈻㧏⏍⏐࡞ࡢࠉ㌹෕โᚒࡓࡄ࡚࡝ࡂࠉ㌹෕ᚃโᚒࠉ⩳ゼᚃࡡโᚒ࠿㔔さ࡚࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡆࡿ࡞ຊ࠻ࠉ┷Ⳟ㢦࡚᭩ኬࡡ࢛ࣜ࢝ࢾ࡚ࣚ࠵ࡾ⣵⬂ባ
ࡡ㏳㐛࠿㔔さ࡚࠵ࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡱࡒࠉᅖమᇰ㣬࡚≁␏Ⓩ࡞㌹෕โᚒࡈࡿ࡙࠷ࡾࢰࣤࣂࢠ㈻࡚ࡵࠉᾦ
మᇰ㣬࡚ᙁโⓆ⌟ࡈࡎࡒሔྙᚪࡍࡊࡵฦἢ⏍⏐ࡈࡿ࡝࠾ࡖࡒ஥࠾ࡼࠉᅖమᇰ㣬࡚ࡢరࡼ࠾ࡡ≁␏Ⓩ࡝ࢰࣤࣂࢠ㈻⏍⏐ᶭᵋ࠿࠵ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
ࡆࡿࡱ࡚ࡡࣈࣞࢷ࢛࣭࣑よᯊࡡ⤎ᯕ࠾ࡼࠉᅖమᇰ㣬࡚ࡢ⏍⏐ࢰࣤࣂࢠ㈻࡛ࡊ࡙⣵⬂හࡡࢰࣤࣂࢠ㈻࡛ᛦ
ࢂࡿࡾࡵࡡ࠿୹さ࡝ࢪ࣎ࢴࢹ࡛ࡊ࡙᳠ฝࡈࡿࡾ࡝࡜ࠉᾦమᇰ㣬࡚ࡡࢰࣤࣂࢠ㈻⏍⏐ࡷࠉ㓕ẍ➴௙ࡡ┷Ⳟ㢦࡚ࡢሒ࿈ࡈࡿ࡙࠷࡝࠷⌟㇗ࡵぜࡼࡿ࡙࠷ࡾࠊᠻࠍࡢࠉࡆࡿࡼࡡ⌟㇗࡞ࡗ࠷࡙ࠉ࣏࢕ࢠࣞ࢓ࣝ࢕よᯊࡵྱࡴࡒฦᏄ⏍∸ᏕⓏ࡝よᯊ࡞ࡻࡽよ᪺ࡌࡾࡆ࡛ࢅ┘Ⓩ࡞ࠉ⌟ᅹ◂✪ࢅ㐅ࡴ࡙࠷ࡾࠊࡱࡒࠉᚋࡼࡿࡒ▩ぜࢅᇱ࡞ࠉ㯔Ⳟᰬ࠵ࡾ࠷ࡢࢰࣤࣂࢠ㈻⏍⏐᪁ἪࡡᨭⰃࢅモࡲ࡙࠷ࡾࠊ
㸟ᮇ◂✪ࡢࠉ⏍◂ࢬࣤࢰ࣭ᇱ♇◂✪᥆㐅஥ᴏࠔࢰࣤࣂࢠ㈻ᕝሔ࡛ࡊ࡙ࡡ⣊≟Ⳟࡡ㧏ᗐฺ⏕࡞㛭ࡌࡾᇱ
┑Ⓩ◂✪ࠕࡡୌ⎌࡛ࡊ࡙⾔ࢂࡿࡱࡊࡒࠊ
9KHS URWH RPH FQF O^VL VL VW KHW ULJJ HUWRXQ[ HLOW KHSURW HLQ SUR IXHW LRQ PHH KFQLV P 
RI &VS HUJL OOXV RU^ ̲FH 
0F̲XKLUR.\FVKLWF0HQ4IF 3FWLRQFO7HVHFUHK.QVWLWXWHRI'UH\LQJ 

21

S-6
ࢤࢺࣤ᭩㐲໩࡞ࡻࡾ㯔Ⳟࡡ␏⛸ࢰࣤࣂࢠ㈻⏍⏐ຝ⋙ࡡྡྷ୕஫࿝ົஒ㸝᮶໪ኬᏕኬᏕ㝌㎨Ꮥ◂✪⛁㸞㯔Ⳟࢅࡢࡋࡴ࡛ࡌࡾ Aspergillus ᒌ࢜ࣄࡢࠉཿࡂ࠾ࡼ㔂㏸⏐ᴏࡷ㓕⣪๠⏍⏐࡞ኣ⏕ࡈࡿ࡙࠽ࡽࠉ㧏࠷Ꮽධᛮ࡛ࢰࣤࣂࢠ㈻㧏ฦἢ⬗ࢅ᭯ࡌࡾ≁ᚡ࠿╌┘ࡈࡿࠉ᭯⏕࡝␏⛸ࢰࣤࣂࢠ㈻ࡡฦἢ⏍⏐⏕ࡡ᭯᭻࡝ᐙ୹࡛ࡊ࡙᭿ᙽࡈࡿ࡙࠷ࡾࠊࡌ࡚࡞࢓ࣜ࢜ࣛ࣬ࣛࣂ࣭ࢭࡷ࣓࢞ࢨࣤ
࡝࡜ࡡ⤄ᥦ࠻మ㓕⣪࠿ᕝᴏⓏ࡞⏍⏐ࡈࡿࠉࡈࡼ࡞ኣ⛸㢦ࡡ␏⛸ࢰࣤࣂࢠ㈻࡞ࡗ࠷࡙⏍⏐ᛮࡡྡྷ୕࠿モࡲࡼࡿ࡙࠷ࡾࠊࡆࡿࡱ࡚ࡢࠉᙁງ࡝Ⓠ⌟⏕ࣈ࣓࣭ࣞࢰ࣭ࢅฺ⏕ࡊ࡙ኣࢤࣅ࣭ᑙ
ථࡌࡾࡆ࡛࡞ࡻࡖ࡙ࠉ┘Ⓩࡡ㐿ఎᏄࡡ㌹෕㔖ࢅቌᙁࡌࡾࡆ࡛࡚⏍⏐㔖ࢅྡྷ୕ࡈࡎࡻ࠹࡛ࡌ
ࡾຑງ࠿୯ᚨ࡚࠵ࡖࡒ࠿ࠉ㌹෕௧㜾ࡡ㐛⛤࡞࠽ࡄࡾ㌹෕⏐∸ࡡᏭᏽ໩ࡷ⩳ゼࡡຝ⋙໩࡝࡜࡞ࡗ࠷࡙ࡢ࡮࡛ࢆ࡜᳠ゞࡈࡿ࡙ࡀ࡙࠷࡝࠾ࡖࡒࠊୌ᪁ࠉୌ⯙Ⓩ࡝⤄ᥦ࠻ࢰࣤࣂࢠ㈻Ⓠ⌟⣌࡞࠽࠷࡙ࠉⓆ⌟㐿ఎᏄࡡ౐⏕ࢤࢺࣤࢅᐙ୹ࡡࢤࢺࣤ౐⏕㢎ᗐ㸝codon usage㸞࡞᭩㐲໩ࡌࡾࡆ࡛ࡢ⩳ゼ㐛⛤ࢅຝ⋙໩ࡊࠉ⏍⏐㔖ᨭၻ࡞᭯ຝ࡚࠵ࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉ࢜ࣄ
࡞࠽ࡄࡾࢤࢺࣤࡡ᭩㐲໩ຝᯕ࡞㛭ࡌࡾ◂✪ࡢᑛ࡝ࡂࠉ≁࡞ࢰࣤࣂࢠ㈻⏍⏐ࡡᐙ୹࡛ࡊฺ࡙
⏕ࡈࡿ࡙࠷ࡾ Aspergillus ᒌ࡞࠽࠷࡙ࡢࠉ㔕⏍ᆵ㐿ఎᏄ࡛ࡡⓆ⌟㔖ࡡẒ㍉࡝࡜ࡡレ⣵࡝よᯊࢅ⾔ࡖࡒሒ࿈ࡢ࡝࠷ࠊࡐࡆ࡚ࠉⁿ⩽ࡼࡢ A. oryzae ࢅᐙ୹࡛ࡊࡒ␏⛸ࢰࣤࣂࢠ㈻Ⓠ⌟࡞࠽ࡄࡾࢤࢺࣤ᭩㐲໩ࡡຝᯕ࡞ࡗ࠷࡙ㄢ࡬ࡾࡒࡴࠉࢱࢼ࢓ࣝࣜࢣࣤࢰࣤࣂࢠ㈻࡚࠵ࡾ Der f7 ࡡ㯔Ⳟ࡞ࡻࡾⓆ⌟⣌ࢅ࣓ࢸ࡛ࣜࡊ࡙Ⓠ⌟࡝ࡼࡦ࡞ฦἢ⏍⏐㔖ࢅẒ㍉᳠ゞࡊࡒࠊࢤࢺࣤ᭩㐲໩㐿ఎᏄࡡࢸࢧ࢕ࣤࡢࠉࢤࢺࣤ౐⏕㢎ᗐ࡞㛭ࡌࡾࢸ࣭ࢰ࣭࣊ࢪ
(http://www.kazusa.or.jp/codon/)ࢅཤ⩻࡞ࡊ࡙⾔ࡖࡒࠊࡢࡋࡴ࡞ Der f7 ⮤㌗ࡡࢨࢡࢻࣜ㒼า࡞ࡻࡽ├᥃ฦἢࡈࡎࡾࢤࣤࢪࢹࣚࢠࢹ࡞ࡗ࠷࡙ฦἢ㔖ࢅẒ㍉᳠ゞࡊࡒࠊ1 ࢤࣅ࣭ࡡ㐿ఎᏄ࠿ྜྷࡋ᯹Ⰵమ㒂న࡞ᑙථࡈࡿࡒᰬࢅ 2 ᰬࡍࡗ㐽ࡦࠉᾦమᇰᆀ࡚ᇰ㣬ᚃࠉᇰ㣬୕ΰࢅ SDS-PAGE࡞౩ࡊ࡙ࠉࢗ࢘ࢪࢰࣤよᯊ࡞ࡻࡖ࡙⏍⏐㔖ࢅㄢ࡬ࡒ࡛ࡆࢀࠉ㔕⏍ᆵࢤࢺࣤࢅࡵࡗᰬ(ND)࡚
ࡢࡱࡖࡒࡂࢨࢡࢻࣜࢅ᳠ฝ࡚ࡀ࡝࠾ࡖࡒࡡ࡞ᑊࡊ࡙ࠉࢤࢺࣤࢅ᭩㐲໩ࡊࡒᰬ(OD)࡚ࡢ Der
f7 ࡞┞ᙔࡌࡾࣁࣤࢺ࠿᪺▉࡞᳠ฝࡈࡿࡒࠊࡈࡼ࡞ࠉ㯔Ⳟ࡞࠽࠷࡙ຝ⋙Ⓩ࡝ฦἢ⏍⏐࡞ຝᯕ࠿࠵ࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࢡࣜࢤ࢓࣐࣭ࣚࢭ࡛ࡡ⼝ྙࢰࣤࣂࢠ㈻࡛ࡊ࡙Ⓠ⌟ࡈࡎࡾࢤࣤࢪ
ࢹࣚࢠࢹ࡞ࡗ࠷࡙ࡵ᳠ゞࡊࡒ⤎ᯕࠉࢤࢺࣤࢅ᭩㐲໩ࡊࡒ⼝ྙᆵࡡᰬ(GKOD)࡞࠽࠷࡙ Der f7࡞┞ᙔࡌࡾࣁࣤࢺ࠿㟸ᖏ࡞ᙁࡂ᳠ฝࡈࡿࡒ࠿ࠉࢤࢺࣤࢅ᭩㐲໩ࡊ࡙࠷࡝࠷ᰬ(GKND)࡚ࡢᠺ
⇅ᆵࡡ Derf 7 ࡡฦἢ㔖ࡢᑛ࡝࠾ࡖࡒࠊࡆࡡ࡛ࡀࡡࢗ࢘ࢪࢰࣤよᯊࡡ⤎ᯕ࠾ࡼࠉDer f7 ࡡฦἢ㔖ࡢࢤࢺࣤ᭩㐲໩࡞ࡻࡽ⣑ 2㹳5 ಶᨭၻࡈࡿࡒࡆ࡛࠿ฦ࠾ࡖࡒࠊࡆࡡࡻ࠹࡞ࠉࢤࢺࣤࡡ᭩
㐲໩ࡢࠉDer f7 ࢅ├᥃Ⓠ⌟ࡈࡎࡾሔྙࡓࡄ࡚࡝ࡂࠉ⼝ྙࢰࣤࣂࢠ㈻࡛ࡊ࡙Ⓠ⌟ࡈࡎࡾሔྙ
࡞ࡵࠉฦἢ㔖ࡡᨭၻ࡞ᑊࡊ࡙᭯ណ࡝ຝᯕࢅ♟ࡌࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ
ḗ࡞ࡆࡿࡼࡡᙟ㈻㌹ᥦమ࡞ࡗ࠷࡙ Der f7 mRNA 㔖ࡡẒ㍉ࢅ⾔ࡖࡒ࡛ࡆࢀࠉࢤࢺࣤ᭩㐲໩࡞ࡻࡽ㌹෕⏐∸㔖࠿ቌຊࡊ࡙࠷ࡾࡆ࡛ฦ࠾ࡖࡒࠊࢤࢺࣤࢅ᭩㐲໩ࡌࡾࡆ࡛࡚㌹෕⏐∸㔖࡞ኬࡀ࡝ᕣ␏࠿⏍ࡋࡾࡆ࡛ࡢ஢᝷አ࡚࠵ࡖࡒ࠿ࠉࢤࢺࣤ᭩㐲໩࠿㌹෕⏐∸㔖ࡡቌຊࢅᘤࡀ㉫ࡆࡌ࡛࠷࠹ౚࡢ௙࡞ࡵሒ࿈ࡈࡿ࡙࠽ࡽࠉࢤࢺࣤ࠿⩳ゼࡡ༟న࡚࠵ࡾ࡞ࡵ࠾࠾ࢂࡼࡍࠉ࡝ࡏ
ᐙ୹࡫ࡡ᭩㐲໩࠿㌹෕⏐∸ࡡቌຊࢅᘤࡀ㉫ࡆࡌ࠾ࡢ୘᪺࡚࠵ࡾࠊࡐࡆ࡚ᠻࠍࡢ├᥃Ⓠ⌟ᆵࡡࢤࣤࢪࢹࣚࢠࢹࢅ⏕࠷࡙ࠉ㔕⏍ᆵ࡛᭩㐲໩ᆵ࡞࠽ࡄࡾࠉⓆ⌟ㄇᑙ᫤ࡡ Der f7 mRNA ࡡ⵫✒㏷ᗐࢅẒ㍉ࡊࡒࠊࡐࡡ⤎ᯕࠉㄇᑙิ᭿࠾ࡼ⵫✒㏷ᗐ࡞ᕣ࠿⏍ࡋ࡙࠽ࡽࠉmRNA ࡡྙᠺ㏷
ᗐ࡞ᕣ࠿࠵ࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊୌ᪁ࠉmRNA ฦよ࡞ࡢ⩳ゼ࡛භᙲࡊࡒ⤊㊪࠿Ꮛᅹࡌࡾ
ࡆ࡛࠿▩ࡼࡿ࡙࠽ࡽࠉ≁࡞⤂Ḿࢤࢺࣤࡡ࡝࠷ mRNA ࡷࢻࣤࢬࣤࢪን␏ࢅࡵࡗ mRNA ࡢࠉ␏ᖏ࡝ mRNA ࡛ࡊ࡙ࠉ⩳ゼ࡛භᙲࡊࡒ⤊㊪ࡡୌࡗ࡚ฦよࡈࡿࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊࡐࡆ
࡚ࠉDer f7 ࡞ྱࡱࡿࡾࣝ࢓ࢤࢺࣤࡻࡾ⩳ゼຝ⋙ࡡ఩ୖ࠿ࡆࡡࡻ࠹࡝⤊㊪࡞ࡻࡾ mRNA ࡡฦよࢅᘤࡀ㉫ࡆࡊ࡙࠷ࡾྊ⬗ᛮࢅ᳠ッࡌࡾࡒࡴࠉ࢞ࣔࣛ࢓ࡡࢡࣜࢤ࢓࣐࣭ࣚࢭࡢ⩳ゼࡈࡿࡾ
࠿ࠉDer f7 ࡡ⩳ゼ࠿⾔ࢂࡿ࡝࠷ࡻ࠹࡞ࠉ㔕⏍ᆵ⼝ྙࢰࣤࣂࢠ㈻ࡡ GKND ࡡ㏭୯࡞⤂Ḿࢤࢺ22ࣤࢅᑙථࡊࡒࢤࣤࢪࢹࣚࢠࢹ(GKND*)ࢅష⿿ࡊࠉDer f7 㒂ฦࡡ⩳ゼ࠿ࡵࡒࡼࡌⓆ⌟࡫ࡡᙫ㡢ࢅㄢ࡬ࡒࠊࡐࡡ⤎ᯕࠉGKND ᰬ࡛Ẓ㍉ࡊ࡙ GKND*ᰬ࡚ࡢ Der f7 ⼝ྙ㐿ఎᏄࡡ㌹෕⏐∸㔖ࡡ᭯ណ࡝ቌຊ࠿ふᐳࡈࡿࠉ㔕⏍ᆵ Der f7 ࡡ⩳ゼ㐛⛤࠿㌹෕⏐∸㔖ࡡ఩ୖ࡞㛭୙ࡊ࡙࠷ࡾ
ྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊࡊ࠾ࡊࠉቌຊ㔖ࡢࢤࢺࣤ᭩㐲໩ᆵࡡ GKOD ᰬࡡ Der f7 mRNA ࣝ࣊ࣜࡡ༖ฦ࡮࡜࡚࠵ࡾࡆ࡛࠾ࡼࠉࢤࢺࣤ᭩㐲໩࡞ࡻࡾ mRNA 㔖ࡡቌຊࡢ๑㏑ࡡ㌹෕㏷ᗐ࡞ཀྵ࡯
ࡌᙫ㡢࡝࡜ࡵྱࢆࡓ々ྙⓏ࡝ຝᯕࡡ⤎ᯕ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡒࠊ
ࢤࢺࣤ᭩㐲໩ࡢ㯔Ⳟࡡ␏⛸㐿ఎᏄⓆ⌟⣌࡚ࡵ㟸ᖏ࡞᭯ຝ࡚࠵ࡽࠉᐁ㝷࡞⏍⏐㔖ᨭၻࡡࡒࡴࡡᡋ␆ࡡୌࡗ࡛ࡊฺ࡙⏕ࡈࡿࡗࡗ࠵ࡾࠊࡊ࠾ࡊࠉࢤࢺࣤࡡ᭩㐲໩࡚ࡢ㐿ఎᏄࡡධྙᠺ࠿ᚪさ࡝ࡒࡴࠉ≁࡞ኬࡀ࡝ࢰࣤࣂࢠ㈻ࢅ⏍⏐ࡈࡎࡻ࠹࡛ࡊࡒሔྙࡡᡥ㛣࡛ࢤࢪࢹࢅ⩻࠻ࡾ࡛ࠉỏ⏕ᛮ࠿࠵ࡾᡥἪ࡛ࡊฺ࡙⏕ࡊ࡞ࡂ࠷ࡆ࡛ࡢྫྷࡴ࡝࠷ࠊ⌦᝷Ⓩ࡞ࡢ㐿ఎᏄࡡධྙᠺࢅ௒ࡈ
࡝࠷᪁Ἢ࠿᭻ࡱࡊ࠷࠿ࠉࢤࢺࣤ᭩㐲໩ຝᯕࡡᶭᵋࡡよ᪺ࢅ㐅ࡴࡾࡆ࡛࡞ࡻࡽࠉ㔕⏍ᆵࢤࢺ࡚ࣤࡵ᭩㐲໩ࢤࢺ࡛ࣤྜྷࡋຝ⋙࡚␏⛸ࢰࣤࣂࢠ㈻ࢅⓆ⌟࡚ࡀࡾᐙ୹ࢅ㏸ᠺࡌࡾࡒࡴࡡᡥ᤻
࠾ࡽ࠿ᚋࡼࡿࡾࡵࡡ࡛᭿ᙽࡈࡿࡾࠊ

23

S-7
Aspergillus ᒌ⣊≟Ⳟࡡࢰࣤࣂࢠ㈻ኬ㔖⏍⏐ࢨࢪࢷ࣑㸯㧏㌹෕࣬㧏⩳ゼ⣌ࡡ㛜Ⓠᖶ⏛ ᪺⏍㸝ኬ㛭⥪◂㸞㯔Ⳟ㸝Aspergillus ᒌ㸞ࡢࠉࢂ࠿ᅗ࡞࠽࠷࡙༐ᖳࢅ㉰࠻ࡾ㛏࠷ᖳ᭮࡞ࢂࡒࡖ࡙㒿㢦ࡷ㔂㏸㣏ဗࡡ⿿㏸࡞౐⏕ࡈࡿ࡙࠽ࡽࠉ㯔ⳞⳞమ࡛ࡐࡡ⏍⏐∸ࡡᏭධᛮ࠿ࡀࢂࡴ࡙㧏࠷ࡆ࡛ࠉࡈࡼ࡞Ⳟమአ࡞ኣ㔖ࡡ㓕⣪ࢰࣤࣂࢠ㈻ࢅ
ฦἢ⏍⏐ࡌࡾ⬗ງࢅ᭯ࡊ࡙࠷ࡾࡆ࡛࠾ࡼࠉ␏⛸ࢰࣤࣂࢠ㈻⏍⏐ࡡ᭯᭻࡝ᐙ୹࡛ࡊ࡙᭿ᙽࡈࡿ࡙࠷ࡾࠊᠻࠍࡢࠉ
Aspergillus ᒌ࡞࠽࠷࡙┘Ⓩࡡࢰࣤࣂࢠ㈻ࢅຝ⋙ࡻࡂ⏍⏐ࡈࡎࡾࡒࡴ࡞Ⓠ⌟࣊ࢠࢰ࣭ࡡ㛜Ⓠࢅ⾔ࡖ࡙ࡀࡒࠊᮇ
ㅦⁿ࡚ࡢࠉࡱࡍ㌹෕㔖ࢅ㧏ࡴࡾࡒࡴࡡࢨࢪ࢙࣒࣬ࣝࣤࢹ(Region III)ࢅฺ⏕ࡊࡒ㧏Ⓠ⌟ࣈ࣓࣭ࣞࢰ࣭ࡡ㛜Ⓠ࡞
ࡗ࠷࡙ࠉḗ࡞ࠉ㐿ఎᏄࡡⓆ⌟ࢅ㌹෕ᚃ࡚ࣝ࣊ࣜ㧏ࡴࡾモࡲ࡛ࡊ࡙ࠉ5’-㟸⩳ゼ㡷ᇡ(5’UTR)ࡡᨭን࡞ࡻࡾ㧏ຝ⋙⩳ゼ⣌ࡡ㛜Ⓠ࡞ࡗ࠷࡙⣺௒ࡊࡒ࠷ࠊ
㸦㸣ࢨࢪ࢙࣒࣬ࣝࣤࢹ Region III ࢅฺ⏕ࡊࡒ㌹෕Ὡ ᛮ໩ ᙁງ࡝㌹෕Ὡᛮࢅᣚࡗࣈ࣓࣭ࣞࢰ࣭ࢅᵋ⠇ࡌࡾ࡞࠵ࡒࡽࠉ࢓࣐࣭ࣚࢭ⣌㐿ఎᏄࡡⓆ⌟โᚒᶭᵋࢅよᯊࡌࡾ㐛⛤࡚Ⓠぜࡊࡒࠉࢨࢪ࢙࣒࣬ࣝࣤࢹ㸝Region III㸞ࢅኣ㔔ᑙථࡌࡾࡆ࡛࡞ࡻࡾ㌹෕Ὡᛮ໩ࢅモࡲࡒࠊ࢓࣐࣭ࣚࢭ⣌㐿ఎᏄࡡⓆ⌟ࡢࠉ࣏ࣜࢹ࣭ࢪࠉࢸࣤࣈࣤ࡝࡜࡞ࡻࡾㄇᑙࢅḿ࡞โᚒࡌࡾ㌹෕ᅄᏄ AmyRp ࡛ࢤࣤࢬࣤࢦࢪ㒼า(CGG-N8-CGG ࡝࡜)ࡡ⤎ྙࠉ1, 2) ཀྵࡦኣࡂࡡ┷ᰶ᚜⏍∸࡞Ꮛᅹࡌࡾᗀᇡ㌹෕ᅄᏄ࡚࠵ࡾ HAP々ྙమ࡛ CCAAT 㒼าࡡ⤎ྙ 3) ࡞ࡻࡽ㌹෕࡚ࣝ࣊ࣜ༝ㄢࡊ࡙Ὡᛮ໩ࡈࡿࡾࠊ㯔Ⳟࡡ α̺ࢡࣜࢤࢨࢱ࣭ࢭ㐿ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭࡞ࡢࠉࡆࡡ㸧ࡗࡡࢨࢪ࢙࣒࣬ࣝࣤࢹ࠿ Region III ࡛࿣ࡣࡿࡾ㡷ᇡ࡞㏻⤾ࡊ࡙Ꮛᅹࡌࡾࠊࡐࡆ
࡚ࠉ㯔Ⳟ(A. oryzae)ᐙ୹࡞࠽࠷࡙ᵕࠍ࡝ࣈ࣓࣭ࣞࢰ࣭࡞ Region III ࡡ 12 ᅂ⧖㏁ࡊ㒼าࢅᑙථࡊࡒ࡛ࡆࢀࠉA.
oryzae ⏜ᮮࡡࢡࣜࢤ࢓࣐࣭ࣚࢭ㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭࡚⣑ 4 ಶ(P-glaA142)ࠉA. niger ⏜ᮮࡡ No8 ࣈ࣓࣭ࣞࢰ࣭࡚
⣑ 6 ಶ(P-No8142)ࡡࣈ࣓࣭ࣞࢰ࣭Ὡᛮࡡቌຊ࠿ふᐳࡈࡿࡒࠊ4) ࡈࡼ࡞᭩㎾ࠉ㯔Ⳟࡡよ⢶⣌㐿ఎᏄࡡ୯࡚ࡵ≁࡞㧏Ⓠ⌟ࡊ࡙࠷ࡾ࢙ࢿ࣭ࣚࢭ㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭࡫ࡡ Region III ᑙථࢅモࡲࡒ࡛ࡆࢀ 30 ಶ௧୕ࡡࣈ࣓࣭ࣞࢰ࣭Ὡᛮ(P-enoA142)ࡡቌຊࢅ♟ࡊࡒࠊ5)௧୕ࡻࡽࠉRegion III ࡡᑙථࡢࣈ࣓࣭ࣞࢰ࣭ࡡ㌹෕Ὡᛮ໩⬗ࢅᨭⰃࡌࡾ
ࡒࡴࡡ㟸ᖏ࡞ຝᯕⓏ࡝ᡥṹ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡾࠊࡱࡒࠉRegion III ࡡᑙථ࡞ࡻࡾࣈ࣓࣭ࣞࢰ࣭ὩᛮࡡᨭⰃࡢࠉ
A. niger ࡷ A. usamii ࢅᐙ୹࡛ࡊࡒሔྙ࡞ࡵ☔ヾࡈࡿ࡙࠽ࡽࠉAspergillus ᒌᐙ୹࡞࠽࠷࡙ᗀࡂᶭ⬗ࡌࡾࡆ࡛࠿஢
᝷ࡈࡿࡾࠊ
㸧㸣5’UTR ࡡᨭን࡞ࡻࡾ⩳ゼࡡຝ⋙໩ ࡈ࡙ࠉ┘Ⓩࡡࢰࣤࣂࢠ㈻ࢅࡻࡽ㧏࠷࡚ࣝ࣊ࣜⓆ⌟ࡈࡎࡾࡒࡴ࡞ࡢࠉ୕࡚㏑࡬ࡒ㌹෕ࡡὩᛮ໩࡞ຊ࠻࡙ࠉ㌹෕ᚃࡡ㐛⛤࡞࠽ࡄࡾࢰࣤࣂࢠ㈻Ⓠ⌟ࡡຝ⋙ࢅᨭⰃࡌࡾࡆ࡛ࡵ㔔さ࡛⩻࠻ࡼࡿࡾࠊࡐࡡࡻ࠹࡝ふⅤ࠾ࡼࠉᠻࠍ
ࡢḗ࡞ࠉ㌹෕⏐∸(mRNA)ࢅࡻࡽ᭯ຝ࡞ฺ⏕ࡌࡾࡒࡴ࡞⩳ゼࡡຝ⋙໩ࢅモࡲࡒࠊ┷ᰶ⏍∸࡞࠽ࡄࡾୌ⯙Ⓩ࡝⩳ゼ㛜ጙᶭᵋ࡛ࡊ࡙ࡢࠉࣛ࣍ࢮ࣭࣑࡞ࡻࡾ mRNA 5’ᮆ❻࠾ࡼࡡࢪ࢞ࣔࣤࢼࣤࢡ
࣓ࢸࣜ࠿ᥞ၌ࡈࡿ࡙࠷ࡾࠊࡐࡆ࡚ࡢࠉmRNA 5’ᮆ❻࠾ࡼ㛜ጙࢤࢺࣤࡱ࡚ࡡ 5’UTR ࠿㔔さ࡝ᙲ๪ࢅᢰࡖ࡙࠽ࡽࠉ
ࡐࡡ㡷ᇡࡡᵋ㏸ࡷ㒼า࠿⩳ゼຝ⋙࡞ኬࡀࡂᙫ㡢ࡌࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊ
ୌ᪁ࠉ㯔Ⳟࢅࡢࡋࡴ࡛ࡌࡾ࢜ࣄࡡ⩳ゼᶭᵋ࡞ࡗ࠷࡙ࡢ࡮࡛ࢆ࡜◂✪ࡈࡿ࡙࠽ࡼࡍࠉ5’UTR ࡛⩳ゼຝ⋙࡛ࡡ㛭౿ࢅᏽ㔖Ⓩ࡞よᯊࡊࡒሒ࿈ࡢ࡝࠾ࡖࡒࠊࡐࡆ࡚ᠻࠍࡢࠉ⩳ゼ࡞࠽ࡄࡾ 5’UTR ࡡᙫ㡢ࢅ᳠ゞࡌࡾࡒࡴ࡞ࠉྜྷ
ୌࣈ࣓࣭ࣞࢰ࣭࡞␏࡝ࡾ 5’UTR ࢅ㏻⤎ࡊ࣭ࣝ࣎ࢰ࣭㐿ఎᏄ⏐∸ࡡὩᛮ࡞ࡻࡽフ౮ࡊࡒ 6)ࠊࡐࡡ⤎ᯕࠉࢤࣤࢹ
࣭ࣞࣜ࣊ࢠࢰ࣭࡛Ẓ㍉ࡊ㸩㹳㸭ಶࡡὩᛮቌຊ࠿☔ヾࡈࡿࡒࠊࢿࢧࣤよᯊࡻࡽ mRNA 㔖ࡢྜྷ➴࡚࠵ࡾࡆ࡛࠾ࡼࠉ
ᚋࡼࡿࡒ㓕⣪ࢰࣤࣂࢠࡡὩᛮ୕᪴ࡢ⩳ゼ࡚ࣝ࣊ࣜ⏍ࡋ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡱࡒࠉ᭩ࡵ㧏࠷⩳ゼຝ⋙ࢅ♟ࡊࡒⓆ⌟ࢤࣤࢪࢹࣚࢠࢹࡡኣࢤࣅ࣭ᑙථᰬ࡞࠽࠷࡙ࡢࠉ⵫✒ࡊࡒ࣭ࣝ࣎ࢰ࣭㐿ఎᏄ⏐∸㸝GUS ࢰࣤࣂࢠ㈻㸞
࠿Ⳟమහධྊ⁈ᛮࢰࣤࣂࢠ㈻ࡡ 50%௧୕ࢅ༥ࡴࡾ࡮࡜࡚࠵ࡖࡒࠊࡆࡿࡼࡡ⤎ᯕ࠾ࡼࠉ5’UTR ࡡᨭን࡞ࡻࡽ⩳ゼ(㛜ጙ)ຝ⋙ࢅ㧏ࡴࡾࡆ࡛࠿┘Ⓩࢰࣤࣂࢠ㈻ࢅ㧏⏍⏐ࡈࡎࡾ୕࡚㟸ᖏ࡞㔔さ࡚࠵ࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊ௧୕ࠉ㌹෕࣬⩳ゼࡡ୦ࣈࣞࢬࢪ࡞࠽࠷࡙Ⓠ⌟ຝ⋙ࢅ㧏ࡴࡾࡒࡴࡡ࢓ࣈ࣭ࣞࢲࢅ⣺௒ࡊࡒࠊࡆࡿࡼࡡᢇ⾙ࢅ24ฺ⏕ࡊ࡙ࠉࡆࡿࡱ࡚࡞ࡈࡱࡉࡱ࡝᭯⏕ࢰࣤࣂࢠ㈻ࡡ㧏⏍⏐࠿㐡ᠺࡈࡿ࡙࠷ࡾࠊࡊ࠾ࡊ࡝࠿ࡼࠉ┘Ⓩ࡛ࡌࡾࢰ
ࣤࣂࢠ㈻㸝㐿ఎᏄ㸞࡞ࡻࡖ࡙ࡢ㧏Ⓠ⌟໩࠿㞬ࡊ࠷ࢢ࣭ࢪࡵ࠵ࡽࠉ஢᝷ࡈࡿࡾさᅄࡵࡈࡱࡉࡱ࡚࠵ࡾࠊ௑ᚃࠉ
ࡐࡿࡼࡡౚࢅレ⣵࡞よᯊࡌࡾࡆ࡛࡞ࡻࡽࠉࡻࡽỏ⏕ᛮࡡ㧏࠷ຝ⋙Ⓩ࡝Ⓠ⌟ࢨࢪࢷ࣑ࡡᵋ⠇࠿᭿ᙽࡈࡿࡾࠊ
ཤ⩻ᩝ⊡1) Petersen, K. L. et. al.: Mol. Gen. Genet., 262, 668 (1999).
2) Gomi, K. et al.: Biosci. Biotechnol. Biochem., 64, 816 (2000).
3) Kato, M.: Biosci. Biotechnol. Biochem., 69, 663 (2005).
4) ᓙ᫤ಆ㈏: ໩Ꮥ࡛⏍∸, 38, 831 (2000).
5) Tsuboi, H. et. al.: Biosci. Biotechnol. Biochem., 69, 206 (2005).
6) Koda, A. et. al.: Appl. Microbiol. Biotechnol., 66, 291 (2004).
High expression system for foreign proteins in Aspergillus species: Efficient transcription and translation
Akio Koda (General Research Laboratory, Ozeki Co., Ltd.)

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S-8
㯔Ⳟࡡኣᙤ࡝∸㈻⏍⏐ࡡࡒࡴࡡ᩺ぞᐙ୹࣊ࢠࢰ࣭⣌ࡡ㛜ⓆỞ▴⏛ ༡ᶖ㸝᭮᰿෗ ⥪◂㸞ỞỞࡢࡋࡴ࡞ 㯔Ⳟ(Aspergillus oryzae)ࡢࠉΰ㒿ࠉ㓲ἔࠉ࿝ძࠉ࿝㓜࡝࡜ࡡࢂ࠿ᅗࡡఎ⤣Ⓩ࡝㔂㏸⏐ᴏ࡞౐⏕ࡈࡿ࡙ࡀࡒࠊࡆࡿࡼࡡ㔂㏸⏐ᴏ࡚ࡢࠉ⡷࣬ኬ㇃࣬㯇࡝࡜ࡡ⾪㟻࡞㯔Ⳟࢅ⏍⫩ࡈࡎࡾᅖమᇰ㣬࡞ࡻࡽࠉ㟸ᖏ࡞ኬ㔖ࡡ࢓࣐࣭ࣚࢭࡷࣈࣞࢷ࢓࣭ࢭࢅฦἢࡈࡎࠉⓆ㓕࡞ᚪさ࡝✈∸ࡡฦよ࡞ฺ⏕ࡊ࡙ࡀࡒࠊࡱࡒ㎾ᖳࡡṽⳞᢇ⾙ࡡⓆᒈ࡞ࡻࡽࠉ㯔Ⳟࡢᾦమᇰ㣬࡚ࡵኬ㔖ࡡ㓕⣪ࢅฦἢࡌࡾࡆ࡛࠿よ᪺ࡈࡿࠉ⏐ᴏ୕᭯⏕࡝㓕⣪࠿㯔Ⳟࡡᾦమᇰ㣬࡚ࡵ౩⤝ࡈࡿ࡙࠷ࡾࠊࡆ
ࡡࡻ࠹࡞ࠉᅖమᇰ㣬ࡵᾦమᇰ㣬ࡵྊ⬗࡝㯔Ⳟࡢࠉࡐࡡ㓕⣪⏍⏐㔖࡛ฦἢງ࠾ࡼࠉ᚜⏍∸ࡡ୯࡚ࡵ∸㈻⏍⏐ᐙ୹࡛ࡊ࡙㟸ᖏ࡞᭯⏕࡚࠵ࡾࡣ࠾ࡽ࡚࡝ࡂࠉ㛏ᖳࡡ㣏⤊㥺࡞⿤ࡘࡄࡈࡿࡒ
Ꮽධᛮ࠾ࡼࠉGRAS(Generally Recognized As Safe)࡞ࡵヾᏽࡈࡿ࡙࠷ࡾࠊࡻࡖ࡙ࠉ㯔Ⳟࡡ EST
࡛ࢣࢿ࣑㒼า࠾ࡼᚋࡼࡿࡾ᭯⏕㓕⣪㐿ఎᏄࢅࠉ㯔Ⳟࢅᐙ୹࡛ࡊ࡙Ⓠ⌟ࡈࡎࡾࡆ࡛࡞ࡻࡽࠉ
Ꮽධ࡝ࢰࣤࣂࢠ㈻ࢅ⏐ᴏⓏ࡞౩⤝࡚ࡀࡾࠊࡐࡆ࡚ࠉ㯔Ⳟࡡຝ⋙Ⓩ࡝∸㈻⏍⏐ࢅྊ⬗࡛ࡌࡾࡒࡴ࡞ࠉᅖమᇰ㣬࡛ᾦమᇰ㣬࡚Ⓠ⌟ࡌࡾ᩺ぞ࡝ࣈ࣓࣭ࣞࢰ࣭ࠉཀྵࡦ᩺ぞ࡝ᐙ୹࣊ࢠࢰ࣭⣌࡝࡜ࡡ㯔Ⳟ∸㈻⏍⏐ࢵ࣭ࣜࢅ㛜Ⓠࡊࡒࡆ࡛ࢅሒ࿈ࡌࡾࠊ
ᅖమᇰ㣬ࠉᾦమᇰ㣬࡚Ⓠ⌟ࡌࡾࣈ࣓࣭ࣞࢰ࣭ 㯔Ⳟࡢᅖమᇰ㣬࡛ᾦమᇰ㣬࠿ྊ⬗࡝᚜⏍∸࡚࠵ࡽࠉ㯔Ⳟ࡚ຝ⋙Ⓩ࡝∸㈻⏍⏐ࢅ⾔࠹ࡒࡴ࡞ࡢࠉࡐࡿࡑࡿࡡᇰ㣬᮪௲࡚ຝ⋙Ⓩ࡞Ⓠ⌟ࡌࡾࣈ࣓࣭ࣞࢰ࣭ࢅཱིᚋࡌࡾࡡ࠿ᚋ➿࡚࠵ࡾࠊ㯔Ⳟࡡᅖమᇰ㣬࡛ᾦమᇰ㣬ࡡ୦ᇰ㣬᮪௲࡚Ⓠ⌟ࡌࡾࣈ࣓࣭ࣞࢰ࣭࡛ࡊ࡙ࡢࠉ᪜࡞ amyB,
glaA ࡝࡜ࡡࣈ࣓࣭ࣞࢰ࣭࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉᠻࠍࡢ㯔Ⳟࡡᅖమᇰ㣬࡛ᾦమᇰ㣬࡚
Ⓠ⌟ࡌࡾࣈ࣓࣭ࣞࢰ࣭࡛ࡊ࡙ࠉࡐࡿࡑࡿࡡᇰ㣬≁␏Ⓩ࡞Ⓠ⌟ࡌࡾ㐿ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭ࢅ
㐽ᢝࡊࠉࡐࡡ∸㈻⏍⏐ᛮ࡞ࡗ࠷࡙フ౮ࡊࡒࠊ 㯔Ⳟࡡᅖమᇰ㣬࡚≁␏Ⓩ࡞Ⓠ⌟ࡌࡾ㐿ఎᏄ࡛ࡊ࡙ࠉࢡࣜࢤ࢓࣐࣭ࣚࢭ㐿ఎᏄ glaB ࢅࢠ࣭ࣞࢼࣤࢡࡊࡒࠊᮇࣈ࣓࣭ࣞࢰ࣭ࡢࠉ࣭ࣝ࣎ࢰ࣭࢓ࢴࢬ࢕࠾ࡼᅖమᇰ㣬࡚≁␏Ⓩ࡞Ⓠ⌟ࡌࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡱࡒ glaB ࣈ࣓࣭ࣞࢰ࣭ࢅࡈࡼ࡞㧏Ⓠ⌟ࡌࡾࣈ࣓࣭ࣞࢰ࣭࡫ᨭⰃࡌࡾࡒࡴ
࡞ࠉࣈ࣓࣭ࣞࢰ࣭よᯊ࠾ࡼྜྷᏽࡈࡿࡒᅖమᇰ㣬࡚ࡡⓆ⌟࡞୘ྊḖ࡝㡷ᇡࢅࠉࢰࣤࢸ࣑࡞㸭ࢤࣅ࣭᣼ථࡊࡒᨭንࣈ࣓࣭ࣞࢰ࣭ࢅᵋ⠇ࡊࡒ⤎ᯕࠉᨭንᚃࡡࣈ࣓࣭ࣞࢰ࣭Ὡᛮࡢᅖమᇰ㣬࡚⣑㸪ಶ࡞୕᪴ࡊࡒࠊ 㯔Ⳟࡡᾦమᇰ㣬࡚≁␏Ⓩ࡞Ⓠ⌟ࡌࡾ㐿ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭ࢅ᥀⣬ࡊࡒࠊࡱࡍิࡴ࡞ΰ㒿㯔ࡡ》ን⌟㇗よᯊࡡࡒࡴࠉ㯔Ⳟࡡ》ንᛮ࡞㛭୙ࡌࡾ melO 㐿ఎᏄࡡⓆ⌟᮪௲ࢅ᳠ゞࡊࡒ⤎ᯕࠉഃ↓࡞ࡵᾦమᇰ㣬≁␏Ⓩ࡞Ⓠ⌟ࡌࡾ㐿ఎᏄ࡚࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡽࠉࡐࡡࣈ࣓࣭ࣞ
ࢰ࣭ࡢᾦమᇰ㣬࡚ࡡ∸㈻⏍⏐࡞ฺ⏕࡚ࡀࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡱࡒ㯔Ⳟࡡ EST ࢸ࣭ࢰࡡⓆ⌟㢎ᗐ᝗ሒ࠾ࡼᾦమᇰ㣬࡚≁␏Ⓩ࡞Ⓠ⌟ࡌࡾ㐿ఎᏄࢅ᢫ฝࡊࠉࡐࡡࣈ࣓࣭ࣞࢰ࣭よᯊࢅ⾔ࡖࡒ⤎ᯕࠉᾦమᇰ㣬࡚≁␏Ⓩ࡞㧏Ⓠ⌟ࡌࡾ sodM ࣈ࣓࣭ࣞࢰ࣭ࢅࢠ࣭ࣞࢼࣤࢡࡌࡾࡆ
࡛࡞ᠺຉࡊࡒࠊ㯔Ⳟࡡ᩺ぞ࡝ᐙ୹࣊ࢠࢰ࣭⣌ࡡ㛜Ⓠ 㯔Ⳟࡡᐙ୹࣊ࢠࢰ࣭⣌ࡡ୯࡚ࡢࠉniaD ࠿ୌ⯙Ⓩ࡞ỏ⏕ࡈࡿ࡙࠷ࡾࠊniaD ࡢ‵ඁᛮ࣏࣭࢜
࣭࡛ࡊฺ࡙⏕ྊ⬗࡚࠵ࡾࡆ࡛࠾ࡼࠉౚ࠻ࡣᅖమᇰ㣬࡚ࡡ㐿ఎᏄ⤄ࡲᥦ࠻⏍⏐࡞㐲ࡌࡾࡻ࠹࡝ࠉ⬂Ꮔᙟᠺ⬗࠿㧏࠷ᰬ࡞ࡗ࠷࡙ࡵᐙ୹࣊ࢠࢰ࣭࡛ࡊฺ࡙⏕࡚ࡀࡾࠊࡊ࠾ࡊࠉ㐽ᢝ࣏࣭࣭࢜࠿ niaD ࡡሔྙࠉᐙ୹࡞ᑙථࡈࡿࡾ㐿ఎᏄࡡࢤࣅ࣭ᩐ࠿ୌ⯙࡞㸦ࢤࣅ࣭࡚࠵ࡽࠉ∸㈻⏍⏐࡞ฺ࡚᭯࡝࠷ࠊࡐࡆ࡚ࠉ⬂Ꮔᙟᠺ⬗࠿㧏࠷㯔Ⳟ࡚᩺ぞ࡝ᐙ୹࣊ࢠࢰ࣭⣌ࡡᵋ⠇ࢅモࡲࡒࠊΰ㒿㯔Ⳟ 1013 ᰬ࡞⣰አ⥲↯ᑏࡊࠉࣞ࢕ࢨࣤさịᛮࢅ♟ࡌᰜ㣬さịᛮን␏ᰬࢅᚋࡒࠊᙔルን26␏ࢅ┞⿭ࡌࡾ㐿ఎᏄࢅ㯔Ⳟࢣࢿ࣑ DNA ࠾ࡼࢨࣘࢴࢹ࢝ࣤࢠ࣭ࣞࢼࣤࢡࡊࡒ⤎ᯕࠉ┞⿭㐿ఎᏄࡢ β-࢕ࢮࣈࣞࣅࣜࣛࣤࢥ㓗ࢸࣃࢺࣞࢣࢻ࣭ࢭࢅࢤ࣭ࢺࡌࡾ leuA ࡚࠵ࡾࡆ࡛࠿ึ᪺ࡊࡒࠊ୕オን␏ᰬ࡞ᑊࡊ࡙ leuA ࢅ⏕࠷࡙ᙟ㈻㌹ᥦࡊࡒ⤎ᯕࠉ1-3 ࢤࣅ࣭ࡡࣈࣚࢪ࣐ࢺ࠿ᑙථࡈࡿ
ࡾࡆ࡛ࢅ☔ヾࡊࠉ᩺ぞ࡝㯔Ⳟᐙ୹࣊ࢠࢰ࣭⣌ࡡᵋ⠇࡞ᠺຉࡊࡒࠊ
ᮇᐙ୹࣊ࢠࢰ࣭⣌ࢅ⏕࠷࡙ࠉᾦమᇰ㣬࡚ࡢ sodM ࣈ࣓࣭ࣞࢰ࣭ࠉᅖమᇰ㣬࡚ࡢ glaB ࣈ࣓ࣞ ࣭ ࢰ ࣭ ࢅ ฺ ⏕ ࡊ ࡒ ∸ ㈻ ⏍ ⏐ ࡞ ࡗ ࠷ ࡙ ᳠ゞ ࡊ ࡒ ࠊ ᾦ మ ᇰ 㣬 ࡚ ࡢ ᇰ 㣬 㸨 ᪝ 㛣 ࡚ ᭩ ኬ
1500mg/L-broth ࡡࢡࣜࢤ࢓࣐࣭ࣚࢭࡡ⏍⏐࠿☔ヾࡈࡿࠉᅖమᇰ㣬࡚ࡢⳞమහ㓕⣪࡚࠵ࡾࣝࢠࢲࣤ࠿Ⳟᑊአ࡞ฦἢࡈࡿࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡻࡖ࡙ࠉ㯔Ⳟࡡᾦమᇰ㣬࡛ᅖమᇰ㣬ࡡཫ᪁࡚ࠉ㯔Ⳟࡡ㐿ఎᏄ⤄ࡲᥦ࠻࡞ࡻࡾ∸㈻⏍⏐ࢅྊ⬗࡛ࡌࡾࢵ࣭ࣜ㛜Ⓠ࡞ᠺຉࡊࡒࠊ
ㅨ㎙ ᮇ◂✪ࢅ⾔࠹࡞࠵ࡒࡽࠉ㯔Ⳟࡡᐙ୹࣊ࢠࢰ࣭⣌ࡡ㛜Ⓠ࡞ࡗ࠷࡙ࡇᣞᑙ࠷ࡒࡓࡀࡱࡊࡒࠉ
≺❟⾔ᨳἪெ㣏ဗ⥪ྙ◂✪ᡜᚺ⏕᚜⏍∸㒂⣊≟Ⳟ◂✪ᐄ ᐄ㛏 ᯵ᮄ ㇇༡ኃ࡞ཉࡂᚒ♡⏞ࡊ୕ࡅࡱࡌࠊ
Development of a novel host-vector system for protein overproduction in Aspergillus oryzae.
Hiroki Ishida
Research Institute, Gekkeikan Sake Co. Ltd.

27

S-9

High level production of biopharmaceuticals in the fungus Aspergillus oryzae.
Carsten M. Hjort, Fungal Gene Technology, Novozymes A/S
The Aspergillus oryzae expression system has been developed by Novozymes and used forrecombinant protein production since 1988. Later on also Aspergillus niger and Fusarium
venenatum expression systems have been developed.
The rationale for entering development of the Aspergillus oryzae expression system was that weneeded an efficient expression system that could secrete large amounts of protein at a high productpurity for production of industrial enzymes.
The choice of Aspergillus oryzae as the preferred expression system was made fromseveral different criteria. First of all Aspergillus oryzae is a well characterized organism that has
been used in the food and fermentation industry in Japan for several hundred years. This means thatthe products of the fungus have been recognized as “Generally Rearded As Safe” (GRAS). The
system was also used by producers of industrial enzymes using submerged fermentation for
production for decades as Aspergillus oryzae has been used for production of amylases andproteases since the dawn of modern enzyme manufacturing. Another important advantage of
Aspergillus oryzae was that it could be genetically manipulated using methods based on therecombinant DNA technology present at the time the project was initiated. In the mid eighties it was
not trivial to develop methods for transformation and genetic manipulation of a new microorganism.
Finally there were no patents blocking the use of Aspergillus oryzae at the time.
The early recombinant strains were essentially wild type strains transformed with plasmids
based on a promoter and a selection marker that was cloned directly from different Aspergillus
strains. Several shortcomings of these strains were quickly realized and work to improve the system
was undertaken.
The host strain produced a lot of unwanted proteins such as amylases, amyloglycosidases andproteases. Proteases are particularly troublesome as they can degrade the enzyme product during
production or severely interfere with the stability of the final product. The production of these
proteins could be abolished by either gene disruption of the corresponding specific gene or bymanipulation of wide domain transcription factors. In Aspergillus niger extracellular expression ofproteases can be abolished almost completely by deletion of one single transcription activator
without any identified pleiotrophic effects.
In addition to unwanted proteins, the Aspergillus expression systems also produce anumber of unwanted metabolites. These are both primary metabolites such as acids and secondary
metabolite. The production of such metabolites can be abolished by either random mutation
followed by screening for the desired mutants or by a rational metabolic engineering approach.
Aspergillus niger produce substantial amounts of oxalic acid. This is a problem both because oxalic
acid is toxic, oxalic acid precipitates with calcium which coarse recovery process problems andproduct quality problems and finally the formation of oxalic acid is simply a waste of carbohydrate
fed to the fermentation. We have identified a gene responsible for the formation of oxalic acid anddisrupted that gene in Aspergillus niger. Metabolic flux analysis of the resulting strain proves thatthe oxalic acid production has been abolished while the flux through the glycolysis and the TCA
cycle remains almost unaltered.
In addition to improving the host strain, the expression vectors have also been improved. In
particular the promoter has been the subject of optimzation.
The promoter used in the first recombinant Aspergillus oryzae production strains was the
TAKA promoter, the promoter driving the expression of the abundant TAKA amylase. We havecharacterized this promoter and found a specific transcription factor called amyR and binding sites

28

for that transcription factor in the TAKA promoter. The detailed understanding of the promoter has
enabled us to design new synthetic promoters that are substantially improved compared to the wild
type TAKA promoter.
The improvements of the Aspergillus oryzae expression system have resulted in a versatile,
clean and high yielding expression system for enzyme production. With these improvements thesystem is suitable for other uses than just expression of enzymes, including production of
bioparmaceuticals. So recently we have initiated a number of projects aiming at developing
biopharmaceutical products using Aspergillus oryzae as expression host.
Expression of monoclonal antibodies in Aspergillus has been suggested by other groups. We have
now developed a heterokaryon system for monoclonal antibody expression where the heavy chain
and the light chain are transformed into two different parent cells and are co-expressed in theheterokaryon cell. A number of the challenges including yield improvement and post translational
modifications of the monoclonal antibody, particularly the N linked glycosylation will be discussed.
We have also used the Aspergillus oryzae system for expression of a number of antimicrobial
peptides as well as other proteins of pharmaceutical interest.
The technical challenges will be discussed and the very significant regulatory challenges
will be touched upon. Also the intellectual property landscape is very complex and will briefly be
introduced.


29

S-10
Genomic analysis of protein secretion and associated stresses in Aspergillus
David B. Archer
School of Biology, University of Nottingham, University Park, Nottingham, NG7 2RD, UK
Email: david.archer@nottingham.ac.uk
This is an auspicious time for Aspergillus genomics. Manuscripts that describe the genome
sequences and annotation for A. nidulans (Galagan et al., 2005), A. fumigatus (Nierman et al., 2005)
and A. oryzae (Machida et al., 2005) have been submitted for publication. The genome of A. niger
has

been

sequenced(see

Archer

&

Dyer,

2004)

and

that

sequence(http://www.dsm.com/dfs/innovation/genomics/) is being annotated by the company thatcommissioned the sequencing, DSM, and an invited group of experts. The intention is that a
description of the annotated sequence will be published in 2006. In addition, the genome of anotherstrain

of

A.

niger

is

being

sequenced

by

the

US

DepartmentofEnergy(http://www.microbialgenome.org/announcement/seq2005.shtml) and the genome of A. flavus is
also

being

sequenced

and

the

annotatedversion

will

soon

be

accessible(http://www.aspergillusflavus.org/Genomics.html). A useful web-site with links to all the
Aspergillus genome data is http://www.aspergillus.man.ac.uk. Other useful web-sites are referred toin Archer and Dyer (2004). Thus, we will have available to us the genome sequences of a sexual
Aspergillus sp. that has served as a model for genetic studies, A. nidulans, a human pathogen, A.
fumigatus, two commercially-important spp., A. niger and A. oryzae, and a close relative of A.
oryzae that produces aflatoxin, A. flavus. These sequences will afford comparative genomic studies(Galagan et al., 2005) and scientists will be able to apply genomic approaches in their studies ofAspergillus biology (Archer and Turner, 2005). Already, for example, it has been possible to showthat A. fumigatus has the potential for sexuality even if it has not yet been demonstrated to occur(Paoletti et al., 2005). Other asexual Aspergillus spp. may also have sexual potential and, if realised
in practise, this would provide new opportunities for scientific exploration and enable theconstruction of new strains for exploitation.
The secretion of proteins is a core component of the lifestyles of Aspergillus spp and their capacity
for efficient secretion has underpinned their commercial exploitation. The genome sequences will
provide a resource for the discovery of new enzymes (Machida et al., 2005) and might provide
insight into the role of secreted enzymes in the pathogenicity of A. fumigatus (Robson et al., 2005).

30

Genome analysis and the use of transcriptomic techniques is beginning to provide insight into themechanisms of protein secretion and the cellular stress responses, including the unfolded protein
response (UPR), that are induced when Aspergillus spp. are used as cell factories for high-level
enzyme production (Sims et al., 2005).
The presentation will focus on what has already been learnt from the Aspergillus genomes about themolecular basis of protein secretion and associated stresses. Repression under endoplasmic
reticulum (ER) stress (RESS) (Pakula et al., 2003; Al-Sheikh et al., 2004) will be discussed inaddition to the UPR, as will transcriptomic studies in A. niger using Affymetrix GeneChips(Guillemette et al., in preparation). Stress is induced either by chemicals (e.g. DTT or tunicamycin)
added exogenously to cell cultures or by the secreted production of heterologous proteins. But, not
all heterologous proteins elicit stress to the same extent and it is not known why some proteins are
secreted well whereas others are poorly-secreted and induce the UPR. ER stress is initiated due topoorly-folded proteins being detected in the lumen of the ER, indicating that the assisted folding (bychaperones and foldases) of heterologous proteins presents a difficulty to the cell. The secreted
production of human lysozymes, which have well-defined folding characteristics (Dumoulin et al.,
2003; Kumita et al., 2005) from fungi may provide insights into defining some predictive rules on
the efficiency of protein folding in the ER and secretion, and this will be discussed.
Acknowledgements
I thank the Biotechnology and Biological Sciences Research Council and DSM for support.
References
Al-Sheikh, H. M., Watson, A. J., Lacey, G. A., Punt, P., MacKenzie, D. A., Jeenes, D. J., Pakula, T.,
Penttilä, M., Alcocer, M. J. C. & Archer, D. B. (2004). Endoplasmic reticulum stress leads to theselective transcriptional down-regulation of the glucoamylase gene in Aspergillus niger.
Molecular Microbiology, 53, 1731-1742.
Archer, D. B. & Dyer, P. S. (2004). From genomics to post-genomics in Aspergillus niger. Current
Opinion in Microbiology, 7, 499-504.
Archer, D. B. & Turner, G. (2005). Genomics of protein secretion and hyphal growth inAspergillus. In The Mycota XIII, Fungal Genomics, ed. A. J. Brown, Berlin: Springer-Verlag, inpress.
Dumoulin, M., Last, A.M., Desmyter, A., Decanniere, K., Canet, D., Larsson, G., Spencer, A.,
Archer, D.B., Muyldermans, S., Wyns, L., Redfield, C., Matagne, A., Robinson, C.V. & Dobson,
C.M. (2003). A camelid antibody fragment inhibits the formation of amyloid fibrils by humanlysozyme. Nature, 424, 783-788.
Galagan, J. E., Calvo, S. E., Cuomo, C. et al. (2005). Sequencing and comparative analysis ofAspergillus nidulans. Nature, submitted.
Kumita, J. R., Johnson, R. J. K., Alcocer, M. J., Dumoulin, M., Holmqvist, F., McCammon, M. G.,
Robinson, C. V., Archer, D. B., & Dobson, C. M. (2005). Expression and characterization of theamyloidogenic variants F57I and W64R of human lysozyme. Journal of Biological Chemistry,
submitted.

31

Machida, M., Asai, K., Sano, M., et al. (2005). Genome sequencing and analysis of Aspergillus
oryzae. Nature, submitted.
Nierman, W., Pain, A., Anderson, M.J., et al. (2005). Genomic sequence of the pathogenic andallergenic Aspergillus fumigatus. Nature, submitted.
Pakula, T. M., Laxell, M., Huuskonen, A., Uusitalo, J., Saloheimo, M. & Penttilä, M. (2003). The
effects of drugs inhibiting protein secretion in the filamentous fungus Trichoderma reesei:
evidence for down-regulation of genes that encode secreted proteins in the stressed cells. Journalof Biological Chemistry, 278, 45011-45020.
Paoletti, M., Rydholm, C., Schweier, E. U., Anderson, M.J., Szakacs, G., Lutzoni, F., Debeaupuis,
J.-P., Latgé, J.-P., Denning, D.W. & Dyer, P.S. (2005), Evidence for sexuality in theopportunistic fungal pathogen Aspergillus fumigatus. Current Biology 15, 1242-1248.
Robson, G. E., Huang., J., Wortman, J. & Archer, D. B. (2005). A preliminary analysis of theprocess of protein secretion, and the diversity of putative secreted hydrolases encoded inAspergillus fumigatus: insights from the genome. Medical Mycology Supplement 1, 43,
S41-S47.
Sims, A.H., Gent, M.E., Lanthaler, K., Dunn-Coleman, N.S., Oliver, S.G. & Robson, G.D. (2005).
Transcriptome analysis of recombinant protein secretion by Aspergillus nidulans and theunfolded protein response in vivo. Applied and Environmental Microbiology, 71, 2737-2747.

32

S-11
ᏄᄙⳞ࠽ࡻࡦ୘ᏰධᏄᄙⳞࡡஹ㒼࡛ஹ㒼ᆵ㐿ఎᏄ᭯Ờ ງ࣬㔘Ꮔ ຉ 1࣬㔘᲻ḿᶖ 2࣬ຊ⸠ࣀࢻ 3࣬஬ᒜ┷⌾ 4࣬ᐣᶌຊዄ࣬ᑈᒱ ᚥ᮶ா㎨ᕝኬᏕኬᏕ㝌࣬1 ⌟ ㎨ᴏ⎌ሾᢇ⾙◂✪ᡜ࣬2 ⌟ ࢦࣤࢹ࣭ࣛ඙㐅ᢇ⾙ᚺ⏕◂✪ᡜ࣬3 ⌟TIS࣬4 ⌟ JICA
ᏄᄙⳞ㛓࡞ᒌࡌࡾ⛸࡞ࡢᇱᮇⓏ࡞ 2 ⛸㢦ࡡஹ㒼ᆵ㸝MAT1-1 ࠵ࡾ࠷ࡢ MAT1-2 ࡡ࠷ࡍࡿ࠾㸞ࡡⳞᰬ࠿Ꮛᅹࡊࠉ␏࡝ࡾஹ㒼ᆵⳞᰬ࠿ஹ㒼ࡌࡾࡆ࡛࡚᭯ᛮ⏍Ṣࢅ⾔࠷ࠉᏰධୠ௥ࢅᙟ
ᠺࡌࡾ㸝self-incompatible = heterothallic㸞ࠊ྘Ⳟᰬࡡஹ㒼ᆵࡢࠉࢣࢿ࣑୕ࡡஹ㒼ᆵ㐿ఎᏄ
㸝MAT1㸞㡷ᇡ࡞Ꮛᅹࡌࡾᑊ❟㐿ఎᏄ⩄㸝MAT1-1 㐿ఎᏄ⩄࠵ࡾ࠷ࡢ MAT1-2 㐿ఎᏄ⩄㸞ࡡ࠷ࡍࡿࢅᣚࡗ࠾࡞ࡻࡖ࡙Ửᏽࡈࡿࡾࠊ
MAT1 㡷 ᇡ ࡡ よ ᯊ ࡢ ࠉ ᙔ ิ ࠉ Neurospora crassa ࠉ Podospora anserina ࠉ Cochliobolus
heterostropus㸝ࢹ࣓ࢗࣞࢤࢨࡇࡱⴝᯜ⑋Ⳟ㸞➴࡚㐅ࡴࡼࡿࡒࠊ࠷ࡍࡿࡡ⛸࡞࠽࠷࡙ࡵ MAT1
㡷ᇡ࡞ࡢࠉMAT1-1ࠉMAT1-2 Ⳟᰬ㛣࡚ሲᇱ㒼าࡡ┞ྜྷᛮ࠿ヾࡴࡼࡿ࡝࠷㡷ᇡ࠿ࠉ୦ஹ㒼ᆵ
Ⳟᰬ࡚භ㏳ࡡ㡷ᇡ࡞ᣫࡱࡿ࡙Ꮛᅹࡌࡾࡆ࡛࠿ሒ࿈ࡈࡿࡒࠊࡆࡡ㟸┞ྜྷ㡷ᇡࢅࡐࡿࡑࡿ
MAT1-1ࠉMAT1-2 ࢕ࢸ࢔࢛࣓ࣜࣆ㸝idiomorph㸞࡛࿣ࡩࠊ࢕ࢸ࢔࢛࣓ࣜࣆ୕࡞ࡢࠉࡐࡿࡑࡿࠉ
DNA ⤎ྙࢺ࣒࢕ࣤ㸝α-box ࠽ࡻࡦ HMG-box㸞ࢅᣚࡗ MAT1-1-1 ࠽ࡻࡦ MAT1-2-1 㐿ఎᏄ࠿Ꮛᅹࡌࡾࠊࡐࡿࡑࡿࡡ DNA ⤎ྙࢺ࣒࢕ࣤࡡ࢓࣐ࢿ㓗㒼าࡢ⛸ࢅ㉰࠻࡙ಕᏋᛮ࠿㧏࠷ࡡ࠿≁ᚡ࡚࠵ࡽࠉࡆࡿࢅฺ⏕ࡊ࡙ኣ⛸࡚ MAT1 㡷ᇡࡡよᯊ࠿㐅ࢆࡓࠊ㐿ఎᏄ◒ቪᐁ㥺➴࡞ࡻࡖ
࡙ࠉMAT1-1-1 ࠽ࡻࡦ MAT1-2-1 ࠿ࢤ࣭ࢺࡌࡾࢰࣤࣂࢠ㈻࠿ࠉⳞᰬࡡஹ㒼ᆵỬᏽࠉஹ㒼㛜ጙࡡโᚒ࡞㛭୙ࡌࡾࡆ࡛࠿♟ࡈࡿ࡙࠷ࡾࠊCo. hererostrophus ࡡ MAT 㡷ᇡ࡞ࡢࡆࡡ 1 ⤄ࡡ㐿ఎᏄࡡࡲ࠿ࠉN. crassa ࠽ࡻࡦ Po. anserina ࡚ࡢࠉMAT1-1 ࢕ࢸ࢔࢛࣓ࣜࣆ୕࡞ࡈࡼ࡞ 2 ࡗࡡ㐿ఎᏄ MAT1-1-2ࠉMAT1-1-3 ࠿Ꮛᅹࡌࡾࠊࡆࡿࡼࡡ㐿ఎᏄ⏐∸ࡢ⛩ᛮ➴ࠉ᭯ᛮ⏍Ṣ࡞㛭୙ࡌࡾࡆ࡛࠿♟ြࡈࡿ࡙࠷ࡾࠊ
ࡐࡡᚃࠉMycosphaerella graminicolaࠉDidymella rabieiࠉCryphonectria parasiticaࠉGibberella
fujikuroiࠉPaecilomyces tenuipesࠉPyrenopeziza brassicae ➴ࡡ self-incompatible ⛸ࠉ࠽ࡻࡦ Co.
homomorphusࠉG. zeae ➴ࡡ self-compatible 㸝= homothallic㸞⛸࡚ MAT1 㡷ᇡࡡᵋ㏸࠿⢥ᰕࡈࡿ࡙࠷ࡾࠊレࡊࡂࡢ Debuchy & Turgeon 2005 ࢅཤ↯࠷ࡒࡓࡀࡒ࠷ࠊ
ᏄᄙⳞࡢኣࡂࡡஹ㒼୘ධᛮ㸝asexual㸞ࡡ⛸ࢅྱࡳࠊᏰධୠ௥ᙟᠺ࠿ᮅሒ࿈࠾࠵ࡾ࠷ࡢṜ
࡜ぜࡼࡿ࡝࠷ࠉ࠷ࢂࡹࡾ୘ᏰධᏄᄙⳞ㢦࡚࠵ࡾࠊ᳔∸⑋ཋⳞࢅྱࡳ Fusarium oxysporumࠉ
Alternaria alternataࠉBiporalis sacchari ࡷ㯔Ⳟ Aspergillus oryzae ࡚ࡢᏰධୠ௥࠿ᮅሒ࿈࡚࠵
ࡾࠊᙔิࠉࡆࡿࡼࡡ⛸࠿ஹ㒼୘ධ࡚࠵ࡾཋᅄ࠿ MAT1 㡷ᇡࡡḖ᥾ࡷ୘ධࠉୌ᪁ࡡஹ㒼ᆵⳞᰬࡡḖዯ࡞࠵ࡾࡡ࡚ࡢ࡝࠷࠾࡛᝷ᏽࡈࡿࡒ࠿ࠉࡐࡡᚃ F. oxysporumࠉA. alternataࠉB. sacchari࡞࠽࠷࡙ࡵ୦ஹ㒼ᆵࡡⳞᰬ࠿Ꮛᅹࡌࡾࡆ࡛࠿ DNA ࡚ࣝ࣊ࣜ☔ヾࡈࡿࠉࡈࡼ࡞ࠉࡐࡿࡼࡡ
MAT1 㡷ᇡ࠿㎾⦍ࡡࣉࢷࣞࢰࣛࢴࢠ⛸࡛┞ྜྷ࡚࠵ࡾࡣ࠾ࡽ࠾ࠉᗑ஋ࡌࡾ MAT1-1-1 ࠽ࡻࡦ
MAT1-2-1 㐿ఎᏄ࠿Ⓠ⌟ࡊࠉᶭ⬗ᛮࢅࡵಕᣚࡊ࡙࠷ࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡆࡡࡒࡴࠉஹ㒼୘ධࡡཋᅄࡢࠉஹ㒼㐿ఎᏄࡡୖὮ࡚ᶭ⬗ࡌࡾࠉஹ㒼㛭㏻ࢨࢡࢻࣜఎ㐡⣌࡞࠵ࡾ࡛᝷ᏽࡈࡿ࡙࠷
ࡾࠊ࢕ࢾࡡ᭩㔔さ⑋ᐐ࡚࠵ࡾ࠷ࡵࡔ⑋ࡡ⑋ཋⳞ Magnaporthe grisea㸝anamorph, Pyricularia
grisea㸞ࡢࠉᮇᮮࣉࢷࣞࢰࣛࢴࢠ࡝ஹ㒼ࢅ⾔࠹ࡢࡍ࡚࠵ࡾ࠿ࠉ୯ᅗ㞴༞࡝࡜࡚᤿㞗ࡈࡿࡒୌ㒂ࡡᰬࢅ㝎ࡀࠉ㏳ᖏࠉஹ㒼⬗ࢅ♟ࡈ࡝࠷ࠊ᪝ᮇ⏐࠷ࡵࡔ⑋Ⳟྜྷኃࡡ࠾ࡄྙࢂࡎ࡞ࡻࡾᏰධ
ୠ௥ᙟᠺࡡሒ࿈ࡢ࡝ࡂࠉ㏳ᖏࠉஹ㒼୘ධᛮ࡚࠵ࡾ࡛⌦よࡈࡿ࡙࠷ࡾࠊࡈ࡙ࠉ࠷ࡵࡔ⑋Ⳟᅗ
හฦ㞫ᰬࢅ౩モࡊよᯊࢅ⾔ࡖࡒ⤎ᯕࠉᠻࠍࡢࠉஹ㒼ᆵ MAT1-1 ࠽ࡻࡦ MAT1-2 ࡡᰬ࠿Ꮛᅹ
ࡌࡾࡆ࡛ࠉᨭࡴ࡙ MAT1-1 Ⳟᰬ࡛ MAT1-2 Ⳟᰬࢅ࠾ࡄྙࢂࡎ࡙ࡵᏰධୠ௥ࢅᙟᠺࡊ࡝࠷ࡆ
࡛ࠉ᪝ᮇ⏐ᰬࡡ MAT1 㡷ᇡࡡᵋ㏸ࡢࣉࢷࣞࢰࣛࢴࢠ࡝㞴༞ᰬ࡛࡮࡯ྜྷᵕ࡚࠵ࡾࡆ࡛ࠉMAT1
㡷ᇡ୕ࡡ㐿ఎᏄ㸝MAT1-1-1ࠉMAT1-1-2ࠉMAT1-1-3 ࠽ࡻࡦ MAT1-2-1ࠉMAT1-2-2㸞ࡢ᪝ᮇ⏐33

ᰬ࡚ࡵⓆ⌟ࡊ࡙࠷ࡾࡆ࡛ࠉࢅ᪺ࡼ࠾࡞ࡊࡒࠊᵋ㏸よᯊࡡ㐛⛤࡚ぜฝࡊࡒ MAT1-1-3 ࠽ࡻࡦ
MAT1-2-1 ࡢࠉMAT1-1 ࠽ࡻࡦ MAT1-2 ࢕ࢸ࢔࢛࣓ࣜࣆ୕࠾ࡼභ㏳㒼า࡞࠾ࡄ࡙Ꮛᅹࡌࡾ⮾࿝῕࠷ᵋ㏸ࢅ࡛ࡖ࡙࠷ࡒࠊࡐࡡࡒࡴࠉ୦㐿ఎᏄ࠿ࢤ࣭ࢺࡌࡾ⏐∸ࡢࠉኬ㒂ฦࡡ࢓࣐ࢿ㓗࠿
භ㏳࡚ࠉN ᮆഁࡡഷ࠾࡝࢓࣐ࢿ㓗࠿␏࡝ࡾࡆ࡛࡞ࡻࡖ࡙ࠉஹ㒼ᆵ≁␏ᛮࢅᣚࡗࠊࡆࡡ⏐∸ࡡᕣ࠿ᶭ⬗Ⓩ࡞࡜ࡡࡻ࠹࡝ណ࿝ࢅ⏍ࡋࡾࡡ࠾ࠉࡱࡒࠉ୦㐿ఎᏄ࠿⏍ࡋࡒ⫴ᬊ࡞ࡗ࠷࡙ࡢᮅ
▩࡚࠵ࡾࠊࡈࡼ࡞ࠉMAT1-1-3 㐿ఎᏄࡡ୕Ὦ࡞ CT ཬᚗ㒼า (CT)n ࠿Ꮛᅹࡌࡾࡆ࡛ࠉࡆࡡ
CT ࡡཬᚗᩐ࠿Ⳟᰬ࡞ࡻࡖ࡙␏࡝ࡾ㸝ࣉࢷࣞࢰࣛࢴࢠᰬ࡚ኬࡀࡂࠉஹ㒼୘ධᰬ࡚ᑚࡈ࠷㸞ࡆ
࡛ࠉCT ཬᚗᩐࡡኣ࠷ᰬ࡚ MAT1-1-3 ࡡ㌹෕㔖࠿ኣ࠷ലྡྷ࡞࠵ࡾࡆ࡛ࢅぜฝࡊࡒࠊஹ㒼ᆵ㐿ఎᏄ⏐∸ࡢࠉࣆ࣓࢘ࣞࣤࡷࣆ࣓࢘ࣞࣤࣝࢬࣈࢰ࣭࡝࡜ࡡ⏐⏍ࢅโᚒࡊࠉࣝࢬࣈ
ࢰ࣭࠿ུᐖࡊࡒࠔ௙ஹ㒼ᆵⳞᰬࡡᏋᅹࠕ࡛࠷࠹᝗ሒ࠿୔㔖మ G ࢰࣤࣂࢠ㈻ࠉMAP ࢞ࢻ࣭ࢭ⤊㊪࠵ࡾ࠷ࡢ cAMP ⤊㊪ࢅ௒ࡊ࡙ᰶහ࡞ఎ㐡ࡈࡿࠉⳞ⣊⼝ྙࠉᰶ⼝ྙࠉᏄᐁమᙟᠺ࡝࡜ࡡ
ஹ㒼ᣪິ࠿㉫ࡆࡾࡆ࡛࠿᝷ᏽࡈࡿ࡙࠷ࡾࠊᠻࠍࡢࠉࣉࢷࣞࢰࣛࢴࢠ⛸࡚࠵ࡾ Gibberella
fujikuroi㸝anamorph, Fusarium sacchari; ࢦࢹࢗ࢞ࣄࡊࡺ࠹㢄⭁ᩃ⑋Ⳟ㸞࡞࠽࠷࡙ G ࢰࣤࣂࢠ㈻βࢦࣇࣗࢼࢴࢹ࠽ࡻࡦ MAP ࢞ࢻ࣭ࢭ㐿ఎᏄࡡ◒ቪᰬࢅࡐࡿࡑࡿషฝࡊࠉ࡜ࡔࡼࡡ◒ቪࡵⳞ⣊ᠺ㛏࡞ࡢᙫ㡢ࢅཀྵ࡯ࡈ࡝࠷ࡆ࡛ࠉ㞜ᛮ୘⛩࡛࡝ࡾࡆ࡛ࠉ⑋ཋᛮ࠿఩ୖࡌࡾࡆ࡛ࢅぜฝࡊࡒࠊ௑ᚃࡢࡆࡿࡼࢅྱࡳ᝗ሒఎ㐡⣌ࡡよᯊࢅ㐅ࡴࠉࣉࢷࣞࢰࣛࢴࢠ⛸࡛ஹ㒼୘ධ⛸ࡡẒ㍉ࢅ⾔࠹ࡆ࡛࡚ஹ㒼୘ධࡡཋᅄࢅ᥀ࡽࡒ࠷࡛⩻࠻࡙࠷ࡾࠊ
ཤ⩻ᩝ⊡Debuchy R and Turgeon B. G. (2005) Mating-type structure, evolution and function in
Euascomycetes. The Mycota ed. by K. Esser; Vol I "Growth, Differentiation and Sexuality" , 2nd edition ed.
by U. Kes and R. Fischer., Springer-Verlag, Heidelberg, chapter 15, in press.

Sexual and asexual development in ascomycetes and mating type genes
Tsutomu Arie, Isao Kaneko1, Masaki Kanamori2, Hana Kato3, Makoto Iyama4, Kana Togashi andTohru Teraoka(Tokyo University of Agriculture and Technology, 1National Institute for Agro-Environmental
Sciences, 2SUNTORY, 3TIS, 4JICA)

34

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MATING TYPE GENES-MEDIATED SEXUAL REPRODUCTION IN FILAMENTOUS
ASCOMYCETES
Sung-Hwan Yun
Division of Life Sciences, Soonchyunhyang University, Asan 336-745, Korea
Fungi capable of sexual reproduction use heterothallic (self-sterile) or homothallic (self-fertile)
mating strategies. In most ascomycetes, a single mating type locus, MAT, with two alternate forms(MAT1-1 and MAT1-2) called idiomorphs, controls mating ability. Structural analyses of MAT
sequences from homothallic and heterothallic Cochliobolus and Gibberella species support the
hypothesis that heterothallism is ancestral. In heterothallic species these alternate idiomorphs residein different nuclei. In contrast, all of homothallic species examined carry both MAT1-1 and MAT1-2
in a single nucleus, usually closely linked. The mechanism of conversion from heterothallism tohomothallism is a recombination event between islands of identity in otherwise dissimilar MAT
sequences. Expression of a fused MAT gene from a homothallic Cochliobolus species confers
self-fertility on a MAT-null strain of a heterothallic species, suggesting that MAT alone is sufficient
to change reproductive life style. Conversely, I asked if a homothallic Gibberella species (G. zeae)
could be made strictly heterothallic by manipulation of MAT. Targeted gene replacement was used
to differentially delete MAT1-1 or MAT1-2 from a wild type haploid MAT1-1;MAT1-2 strain,
resulting in MAT1-1;mat1-2, and mat1-1;MAT1-2 strains that were self-sterile, yet able to cross towild type testers and more importantly, to each other. These results indicated that differential
deletion of MAT idiomorphs eliminates selfing ability of G. zeae, but the ability to outcross isretained. They also indicated that both MAT idiomorphs are required for self fertility.
However, not much is known about the gene regulatory pathway, or genes under the control ofmating-type proteins in filamentous ascomycetes. I have focused on the genes specificallycontrolled by the mating type (MAT) locus, a master regulator of this developmental process in G.
zeae. To do identify them, I employed suppression subtractive hybridization between a G. zeae
wild-type strain Z03643 and its isogenic self-sterile mat1-2 strain T43ΔM2-2. Sequence analysis of
1,000 subtractive cDNA clones revealed 327 unigenes of expressed sequence tags. Both reverse
northern and cDNA microarray analyses using these unigenes confirmed that 171 (52.3%) unigenes
were significantly down-regulated in T43ΔM2-2, suggesting that the expression of these genes
might be critical for sexual development in G. zeae. Among these, 98 could be either manually or
automatically annotated based on known functions of their possible homologs. A high frequency of
the genes similar to those involved in signal transduction pathway, transcriptional regulation,
protein degradation, and cellular differentiation indicates that many regulatory functions are
required during the entire mating processes. Northern blot analysis revealed that all of the genes
examined were down-regulated by MAT1-2, but needed for different stages during sexual
development: specific to perithecial formation, vegetative stage, or no stage specific.

35

S-13
PUTATIVE PHEROMONE RECEPTORS AND SIGNALING OF SEXUAL
DEVELOPMENT IN ASPERGILLUS NIDULANS
Kap-Hoon Han
Dept. of Pharmaceutical Engineering, Woosuk University, Wanju, 565-701, Korea

A homothallic ascomycete, Aspergillus nidulans, which develops both sexual and asexual spores
through complicated morphogenic processes, is a model fungus to study spore formation inmolecular level. Many genes have been identified that are associated with asexual development andthe regulatory network of these genes is well established. A. nidulans also produces sexual sporesdeveloped in closed fruiting bodies, called cleistothecia. Sexual development is greatly affected bycarbon sources and environmental stresses as well as growth conditions. To understand genetic
network and regulatory mechanism(s) of the cellular developmental fate decision in response tovarious environmental factors, several genes, including nsdD which encodes a GATA typetranscription factor regarded as positive regulator of sexual development, have been identified byforward genetic screening using the development-specific characteristics in response toenvironmental stresses. In addition, two genes, gprA and gprB, that are predicted to encode putative
G protein-coupled receptors (GPCRs) similar to fungal pheromone receptors, have beencharacterized to reveal whether the putative pheromone receptors affect fruiting body formation ofthis homothallic fungus. Deletion of gprA or gprB resulted in the production of a few small
cleistothecia carrying a reduced number of ascospores, whereas ΔgprA;ΔgprB eliminatedcleistorhecia formation in homothallic conditions. However, ΔgprA and/or ΔgprB did not affectvegetative growth, asexual sporulation, or even cleistothecia formation in outcross, indicating that
GprA and GprB are specifically required for self-fertilization. Over-expression of nsdD resulted in
the production of barren cleistothecia in the ΔgprA;ΔgprB mutant, suggesting that NsdD canpartially rescue the developmental defects caused by deletion of GPCRs and that GprA/B-mediatedsignaling may involved in activation of other genes required for maturation of cleistothecia.

36

S-14
MATING GENES IN THE ASPERGILLUS ORYZAE GENOME:CAN IT HAVE A SEXUAL LIFE CYCLE?
Nanase Yamamoto1, Praveen Rao Juvvadi1, Jun-ichi Maruyama1, David B Archer2, Katsuhiko Kitamoto1
1
Department of Biotechnology, The University of Tokyo, 1-1-1, Yayoi, Bunkyo-Ku, Tokyo 113-8657, Japan.
2
School of Biology, University of Nottingham, University Park, Nottingham, NG7 2RD, UK

Sexual reproduction in general is regulated by mating-type genes, and filamentous fungi exhibit a diversity ofmating-types. In particular, the Aspergillus species undergoing both asexual and sexual life cycles withhomothallic or heterothallic patterns of breeding not only serve as tools for understanding the basis of sexuality,
but also can be role models for the analysis on switching between asexual and sexual pathways. Critical analysesof the genomic information in the three species of Aspergilli (Aspergillus nidulans, A. fumigatus and A. oryzae) is
expected to provide new insights into the cause and consequence of sexuality in filamentous fungi. With therecent report on sexuality in A. fumigatus1, a supposedly “asexual” species, our understanding on theasexual/sexual habits of filamentous fungi seems to require terser analyses to clearly define their sexual/asexualstates. While in A. nidulans, a homothallic species, sexual state is well known and sex in A. fumigatus is beginningto be understood, not much is known on the closely related species, A. oryzae, which until now is considered topropagate “asexually”. In this report we provide genomic evidences towards understanding sexuality in A. oryzae.
In addition to evidences on the existence of sexual pathway genes in A. oryzae, the expression analysis on thealpha-pheromone precursor and receptor genes will also be discussed.
Mating-type genes (MAT) are generally regarded as the primary regulators of sexual activity, and are present at asingle locus with in the genome. In contrast to A. nidulans, which consists of both the MAT-1and MAT-2 gene loci,a typical characteristic representing its homothallic nature, the genome sequencing of A. oryzae RIB40 strain
revealed the existence of only the MAT-1 gene encoding a protein with the alpha-box motif, prompting aconsideration on its sexual identity. In addition to this, the presence of genes encoding the alpha-pheromone andits receptor (designated as AoppgA and AogprA, respectively), which constitute the basic factors indispensable for
eliciting and sexual response, and also the genes encoding the proteases (Kex1, Kex2 and Ste13) required for theprocessing of the alpha-factor strongly implicated the operation of a sexual pathway in this fungus. However, as
yet we could not locate the gene encoding the a-factor, even though the a-receptor (AogprB) and many genes
encoding the proteins (Rce1, Ste24, Ram2, Ste14, Ste16, and Ste23) responsible for the processing andmodification of the a-factor could be identified in the A. oryzae genome. Moreover, the identification of several
other genes directly/indirectly related to sexual response pathway, such as the velvet (ve) gene, known to induce
sexual response in A. nidulans, the transcription factors nsdD and pro1 required for sexual development andmaturation respectively, and the general components of the MAP kinase pathway clearly indicated the inherent
sexual potential of this filamentous fungus.
Existence of mating-related genes raises a question whether they can function in A. oryzae where sexual cycle
has not yet been found. We chose the pheromone precursor and receptor genes (AoppgA, AogprA and AogprB)
among the other mating-related genes and examined their expression. RT-PCR analyses confirmed that all thethree genes were expressed. Further analysis using various culture conditions revealed that the expression of
AoppgA and AogprA genes was upregulated upon carbon source depletion whereas the expression of AogprB generemained unaltered. This result suggested that recognition of alpha-pheromone by its receptor might play animportant role during carbon source starvation condition. Hence we isolated and analyzed AoppgA and AogprA
genes to reveal the function of alpha-factor in A. oryzae.
AoppgA gene was predicted to encode 103 amino acids with a putative secretory signal sequence and two
repeats of the amino acid sequences (~10 aa). Each repeat is followed by Kex2-processing site (-Lys-Arg-) as inother fungal alpha-pheromones. In S. cerevisiae alpha-pheromone precursor is processed by the proteases such as
Kex2 in the Golgi and excreted out of the cells. For confirmation of processing and secretion of alpha-pheromone
in A. oryzae, AoPpgA-EGFP fusion construct was expressed under the control of amyB promoter and its
localization was examined by fluorescent microscopy. EGFP fluorescence was detected at the septa and in thevacuoles, which was also observed when a secretory protein was fused with EGFP and expressed. Supporting the
microscopic analysis, Western blotting using GFP-antibody suggested the secretion of the processed forms of
AoPpgA into the growth medium.
Alpha-pheromone receptor gene, AogprA is deduced to encode a protein belonging to seven-transmembrane-37spanning G protein-coupled receptor (GPCR) family. In S. cerevisiae alpha-pheromone receptor is sensitized byligand binding and activates the downstream components of the heteromeric G protein and MAPK signalingcascade. For localization analysis, AoGprA was expressed as a fusion with EGFP. Upon fluorescence microscopic
analysis EGFP-fluorescence was detected on the plasma membranes and in the vacuoles. This implied that
AoGprA might be exposed on the cell surface and after pheromone binding may internalize into the vacuoles for
degradation.
To further characterize the function of alpha-pheromone and its receptor, the disruption of the AoppgA and
AogprA genes in A. oryzae NSR13 strain (niaD-, sC-, adeA-) was performed using the adeA marker. Phenotypicvariations of these disruptants will be discussed.
1. Paoletti, M. et al., Evidence for sexuality in the opportunistic fungal pathogen Aspergillus fumigatus. Curr Biol.
2005,15, 1242-8.

38

S-15
Aspergillus nidulans ࡞࠽ࡄࡾ୔ࡗࡡ mitogen-activated protein kinase (MAPK) ⤊㊪᮶໪ኬᏕኬᏕ㝌㎨Ꮥ◂✪⛁ᚺ⏕⏍࿤⛁Ꮥᑍᨯཿᕖ೸ኯ㑳ࠉ㜷㒂ᩏᝃࡢࡋࡴ࡞
 MAPK ࡢ㓕ẍ࠾ࡼဳ஘㢦࡞⮫ࡾ┷ᰶ⏍∸࡞ᬉ㐚Ⓩ࡞ಕᏋࡈࡿࡒࣈࣞࢷ࢕ࣤ࢞ࢻ࣭ࢭ࡚࠵ࡾࠊMAPK ⤊㊪ࡢࠉᠺ㛏ᅄᏄ࣬ࢦ࢕ࢹ࢜࢕ࣤ࣬UV ↯ᑏ࣬ᾈ㏩ᅸࢪࢹࣝࢪ࡝࡜ࡡᵕࠍ࡝አ㒂ๆ⃥࡞ࡻࡖ࡙Ὡᛮ໩ࡈࡿࠉ㌹෕โᚒᅄᏄ࠿ᚺ➽㐿ఎᏄ⩄ࡡⓆ⌟ㄢ⟿ࢅࡌࡾࡆ࡛࡚㐲ว࡝⣵⬂ᚺ➽ࢅᘤࡀ㉫ࡆࡌࠊ┷ᰶ⏍∸ࡡ࣓ࢸࣜ⏍∸࡚࠵ࡾฝⰾ㓕ẍ Saccharomyces cerevisiae ࡞࠽࠷࡙ࡢࠉHOG (High-osmolarity glycerol) ⤊㊪ (Hog1)࣬cell integrity⤊㊪ (Mpk1)࣬mating ⤊㊪ (Fus3)࣬᧻Ⳟ⣊ᙟᠺ⤊㊪ (Kss1) ࡝࡜࠿▩ࡼࡿ࡙࠷ࡾ(1)ࠊ㎾ᖳࡡ EST よᯊࡷࢣࢿ࣑よᯊ࡞ࡻࡽࠉ⣊≟Ⳟ࡞࠽࠷࡙ࡵࡻ࠹ࡷࡂ MAPK ⤊㊪ࡡよᯊ࠿㐅ࡴࡼࡿࡾࡻ࠹࡞࡝ࡖ࡙ࡀࡒࠊᮇㅦⁿ࡚ࡢࠉ࢓ࢪ࣋ࣜ࢟ࣜࢪᒌ࢜ࣄࡡ࣓ࢸࣜ⏍∸࡚࠵ࡾ A. nidulans ࡡ MAPK ⤊㊪࡞ࡗ࠷࡙ࡡࡆࡿࡱ࡚ࡡ▩ぜࢅࣝࣄ࣭ࣖࡌࡾ࡛࡛ࡵ࡞ࠉ㓕ẍࡡ MAPK ⤊㊪࡛ࡡ㢦జⅤཀྵࡦ┞㐢Ⅴ࡞ࡗ࠷࡙⣺௒ࡊࡒ࠷ࠊ
㓕ẍ࡛⣊≟Ⳟࡡ MAPK ⤊㊪MAPK ⤊㊪ࡢࠉ୕Ὦࡡࢬࣤࢦ࣭⤊㊪࡛ୖὮࡡ MAPK ࢜ࢪࢢ࣭ࢺ࠾ࡼᠺࡾࠊMAPK ࢜ࢪࢢ࣭ࢺࡢࠉMAPKࠉMAPK ࢅࣛࣤ㓗໩ࡊ࡙Ὡᛮ໩ࡌࡾ MAPKKࠉMAPKK ࢅࣛࣤ㓗໩ࡊ࡙Ὡᛮ໩ࡌࡾ MAPKKK ࡻࡽᵋᠺࡈࡿ
ࡾ࢞ࢻ࣭ࢭ࢜ࢪࢢ࣭ࢺ࡚࠵ࡾࠊA. nidulans ࡢᄿࡗࡡ MAPKࠉHogA (Hog1 ࢛ࣜࢮࣞࢡ)ࠉMpkA (Mpk1 ࢛ࣜࢮࣞࢡ)ࠉMpkB (Fus3/Kss1 ࢛ࣜࢮࣞࢡ)ࠉMpkC (Hog1 ࢛ࣜࢮࣞࢡ) ࢅᣚࡔࠉHog1 ࢛ࣜࢮࣞࢡ࠿㔔々ࡊ࡙࠷ࡾⅤࡷ Fus3/Kss1 ࢰ࢕ࣈࡡ MAPK ࠿ୌࡗࡊ࠾Ꮛᅹࡊ࡝࠷Ⅴ࡚㓕ẍ࡛␏࡝ࡾࠊ㓕ẍࡢ MAPKKK ࡛ MAPKKࢅᄿࡗࡍࡗᣚࡗ࠿ࠉୌࡗࡡ⤊㊪࡚ᶭ⬗࠿㔔々ࡊ࡙࠷ࡒࡽࠉSte11 MAPKKK ࡡࡻ࠹࡞々ᩐࡡ⤊㊪࡞㛭୙ࡊ࡙࠷ࡾ࢞ࢻ࣭ࢭࡵᏋᅹࡌࡾࠊ࡛ࡆࢀ࠿ࠉA. nidulans ࡢ MAPKKK ࡛ MAPKK ࢅ୔ࡗࡍࡗࡊ࠾ᣚࡒ࡝࠷ࡒࡴࠉ
MAPK ࢜ࢪࢢ࣭ࢺࡢ㓕ẍࡻࡽࡵ༟⣟࡝ᵋᠺࢅࡊ࡙࠷ࡾ࡛஢᝷ࡈࡿࡾࠊA. nidulans ࡡ HOG ⤊㊪ࡢ௧ୖ࡚㏑࡬ࡾ࠿ࠉcell integrity ⤊㊪࡛ mating ⤊㊪࡞ࡗ࠷࡙ࡢ in silico よᯊ(2)ࡷୌ㒂ࡡ㐿ఎᏄࡡよᯊ࠿⾔ࢂࡿ࡙࠷ࡾࠊCell
integrity ⤊㊪ࡡ MpkA MAPK ࡡḖ᥾࡚ࡢᴗᛮᠺ㛏࠿኶ࢂࡿࠉmating ⤊㊪ࡡ SteC MAPKKK ࡢ᭯ᛮ⏍Ṣ࡞ᚪ㡪࡚࠵ࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾ(3, 4)ࠊ
A. nidulans HOG (AnHOG) ⤊㊪ (5)┷ᰶ⏍∸ࡡᾈ㏩ᅸᚺ➽ࡡ◂✪ࡢࠉ㐿ఎᏕⓏよᯊࡡᐖ᪾࡝ฝⰾ㓕ẍ࡞࠽࠷࡙᭩ࡵ㐅ᒈࡊ࡙࠽ࡽࠉHOG ⤊㊪ࡡ࡮࡯ධᐖ࠿よ᪺ࡈࡿ࡙࠷ࡾ (ᅒ A)ࠊHOG ⤊㊪ࡢ୕Ὦࡡ஦ࡗࡡᾈ㏩ᅸࢬࣤࢦ࣭⤊㊪ (Sln1ࠉSho1) ࡛ୖὮࡡHog1 MAPK ࢜ࢪࢢ࣭ࢺ࠾ࡼᵋᠺࡈࡿ࡙࠷ࡾࠊSln1 ⤊㊪ (=஦ᠺฦᛮ᝗ሒఎ㐡⤊㊪) ࡢࠉࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ
Sln1-ࣛࣤ㓗ᇱ௯௒ᅄᏄ Ypd1-ࣝࢪ࣎ࣤࢪ࣭ࣝ࢟ࣖࣝࢰ࣭ Ssk1 ࠿ࠉHis-Asp ࣛࣤ㓗ᇱ࣭ࣛࣝࢅᙟᠺࡊ࡙࠽ࡽࠉ
㧏ᾈ㏩ᅸๆ⃥࡚ Ssk2/Ssk22 MAPKKKs ⤊⏜࡚ୖὮ⤊㊪ࢅὩᛮ໩ࡌࡾࠊࡱࡒࠉSho1 ⤊㊪ࡢࠉ㧏ᾈ㏩ᅸๆ⃥࡚
Pbs2 MAPKK ࠿ Sho1 ࡡ SH3 ࢺ࣒࢕࡛ࣤ⤎ྙࡊࠉࡐࡡ⤎ᯕ࡛ࡊ࡙ Ste11 MAPKKK ⤊⏜࡚ୖὮ⤊㊪ࢅὩᛮ
໩ࡌࡾࠊHOG ⤊㊪ࡡ㐵᩷ࡢ㧏ᾈ㏩ᅸវུᛮࢅᘤࡀ㉫ࡆࡊࠉ㏣࡞ HOG ⤊㊪ࡡᜇᖏⓏ࡝Ὡᛮ໩ࡢ⮬Ṓ࡛࡝ࡾࠊࢣࢿ࣑よᯊ࡞ࡻࡽࠉA. nidulans ࡢ㓕ẍ HOG ⤊㊪┞ྜྷ㐿ఎᏄࢅධ࡙᭯ࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒ (ᅒ B)ࠊ
஦ᠺฦᛮ᝗ሒఎ㐡⣌ᅄᏄ࡚࠵ࡾ TcsBࠉYpdAࠉSskA ࡢࠉࡐࡿࡑࡿ㓕ẍࡡ┞ྜྷ㐿ఎᏄࡡḖ᥾ࢅᶭ⬗┞⿭ࡊࡒࡆ࡛࠾ࡼࠉࡆࡿࡼࡢ A. nidulans හ࡞࠽࠷࡙ Sln1 ⤊㊪࡛㢦జࡡᶭᵋ഼࡚࠷࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡾࠊࡊ࠾ࡊ
࡝࠿ࡼࠉSLN1 ࡛␏࡝ࡽ tcsB ࡡḖ᥾࠿⾪⌟ᆵࢅ♟ࡈ࡝࠾ࡖࡒࡆ࡛࠾ࡼ々ᩐࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭࡡ㛭୙࠿⩻࠻ࡼࡿࡾࠊୌ᪁ࠉsskA ࡷ hogA ን␏మࡓࡄ࡚࡝ࡂࠉSsk2/Ssk22ࠉPbs2 ࡡ࢛ࣜࢮࣞࢡࢅࢤ࣭ࢺࡌࡾ sskBࠉpbsB
ࡡን␏మࡵ㧏ᾈ㏩ᅸ᮪௲࡚⏍⫩఩ୖࢅ♟ࡊࠉHogA MAPK ࡡࣛࣤ㓗໩ࡵふᐳࡈࡿ࡝࠾ࡖࡒࠊࡆࡡࡻ࠹࡞ࠉAnHOG ⤊㊪ࡡὩᛮ໩ࡢ஦ᠺฦᛮ᝗ሒఎ㐡⣌౪ᏋⓏ࡚࠵ࡽࠉMAPKK ࡡὩᛮ໩ᶭᵋࡢࠉSln1 ࡛ Sho1 ࡡ஦ࡗࡡ⤊㊪࠾ࡼὩᛮ໩ࡈࡿࡾ㓕ẍࡡࡐࡿ࡛␏࡝ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊPbsB ࡛ Pbs2 ࡡ࢓࣐ࢿ㓗㒼าࢅẒ㍉ࡊࡒ࡛ࡆࢀࠉPbs2 ࡞Ꮛᅹࡌࡾ Ssk2/Ssk22 ⤎ྙࢺ࣒࢕ࣤࡷ Sho1 ⤎ྙࢺ࣒࢕ࣤ (=Pro-rich motif) ࡢಕᏋࡈࡿ࡙
࠷࡝࠷ࡻ࠹࡞ᛦࢂࡿࡒࠊPbsB ࡢࠉpbs2Δ ࡡ㧏ᾈ㏩ᅸវུᛮࢅ┞⿭ࡊࡒ࠿ࠉPbsB ࠿㓕ẍහ࡚ Ssk2/Ssk22 ࡛
Sho1 ࡡ࡜ࡔࡼ࠾ࡼὩᛮ໩ࡈࡿࡒࡡ࠾࡛࠷࠹␪ၡ࠿⏍ࡋࡒࠊࡐࡆ࡚ࠉ∞᪁ࡡ⤊㊪࠿Ḗ᥾ࡊࡒ pbs2Δ (sho1Δ
pbs2Δࠉssk2Δ ssk22Δ pbs2Δ) ࡞࠽࠷࡙ pbsB ࢅⓆ⌟ࡈࡎࡒ࡛ࡆࢀࠉsho1Δ pbs2Δ ࡡ᪁ࡓࡄࢅᶭ⬗┞⿭ࡊࡒࠊࡆ
ࡿࡼࡡ⤎ᯕ࠾ࡼࠉ㓕ẍහ࡞࠽ࡄࡾ PbsB ࡡὩᛮ໩ࡢ Ssk2/Ssk22 ࡞౪Ꮛࡊ࡙࠽ࡽࠉPbsB ࡢ Sho1 ࡛⤎ྙ࡚ࡀ࡝࠷ࡆ࡛࠿♟ြࡈࡿࡒࠊA. nidulans හ࡞࠽࠷࡙ࠉPbsB ࡢ඼ᆵⓏ࡝ Pro-rich motif ࢅᣚࡒ࡝࠷ࡒࡴ ShoA ࡛⤎ྙ࡚ࡀࡍࠉࡐࡡ⤎ᯕ࡛ࡊ࡙ࠉHogA ࡡὩᛮ໩ࡢ SskA ࡷ SskB ౪ᏋⓏ࡞㉫ࡆࡾ࡛஢᝷ࡊࡒࠊࡐࡆ࡚ࠉPbsB ࡡ
N ᮆ❻㡷ᇡ࡞ Pbs2 ⏜ᮮࡡ Pro-rich motif (KPLPPLPV) ࢅ᣼ථࡊࡒን␏మ PbsB(Pro) ࢅష⿿ࡊࡒࠊPbsB(Pro)ࡢ ssk2 Δ ssk22 Δ pbs2Δ ࡡ㧏ᾈ㏩ᅸវུᛮࢅ┞⿭ࡊࡒࡆ࡛࠾ࡼࠉPbsB(Pro) ࡛ Sho1 ࡡ㐿ఎᏕⓏ┞பష⏕࠿♟ࡈࡿࡒࠊࡊ࠾ࡊ࡝࠿ࡼࠉPbsB(Pro) ࢅ HogA ࢅὩᛮ໩ࡌࡾࡆ࡛࠿࡚ࡀ࡝࠷ sskAΔ ࡞࠽࠷࡙Ⓠ⌟ࡈࡎࡒ࡛ࡆࢀࠉ

39㧏ᾈ㏩ᅸវུᛮࡡᅂᚗࡢぜࡼࡿࡍࠉHogA ࡡὩᛮ໩ࡵふᐳࡈࡿ࡝࠾ࡖࡒࠊࡆࡿࡼࡡ⤎ᯕ࠾ࡼࠉPbsB ࠿ Pro-richmotif ࢅᣚࡒ࡝࠷ࡆ࡛࡞ຊ࠻࡙ࠉShoA ࠾ࡼࡡ㧏ᾈ㏩ᅸᚺ➽ࢨࢡࢻࣜ࠿ PbsB ࡞ఎ㐡࡚ࡀ࡝࠷రࡼ࠾ࡡḖ᥾࠿Ꮛᅹࡌࡾࡵࡡ࡛஢᝷ࡈࡿࡒࠊࡆࡿࡼࡡࡆ࡛ࢅ๑ᥞ࡞ࠉAnHOG ⤊㊪࡛ mating ⤊㊪ࡡ㛭౿࡞ࡗ࠷࡙ࡵ⣺௒ࡊࡒ࠷ࠊཤ⩻ᩝ⊡1) Hohmann S. (2002) Microbiol Mol Biol Rev 66: 300-372.
2) Muthuvijayan V. et al. (2004) In Silico Biol 4: 605-631.
3) Bussink H.J. et al. (1999) FEMS Microbiol Lett 173: 117-125.
4) Wei et al. (2003) Mol Microbiol 47: 1577-1588.
5) Furukawa K. et al. (2005) Mol Microbiol 56: 1246-1261.
Three mitogen-activated protein kinase (MAPK) pathways in Aspergillus nidulans.
Kentaro Furukawa, Keietsu Abe (Tohoku Univ., Grad. Sch. Agri. Sci.)

40

O-1
㯔Ⳟࡡࣆ࢔ࣜࢰ࣭ᇰ㣬᮪௲ୖ࡚ࡡⓆ⌟よᯊఫ㔕ඔ᫓㸡ᑚᯐலᕵᏄࠉ⏣⏛㞖஄ ࠉኬ⟺ಘୌ 㸝㔘Ἁᕝኬࠉ ⏐⥪◂㸞

<┘Ⓩ> ⣊≟Ⳟࡢ⏐ᴏⓏ࡞ࡵ㔔さ࡝᚜⏍∸࡚ࠉᠻᅗ࡚ࡢࠉఎ⤣ⓏⓆ㓕⏐ᴏࡷࢰࣤࣂࢠ㈻ࡡฦἢ⏍⏐࡝࡜࡞㯔Ⳟ Aspergillus oryzae ࠿ᗀࡂฺ⏕ࡈࡿ࡙࠷ࡾࠊ㯔Ⳟࡡᇰ㣬ࡢኬࡀࡂฦࡄ࡙ࠉࣆࣚࢪࢤ࡝࡜࡞ᇰ㣬ᠺฦࢅථࡿ࡙
ᇰ㣬ࢅ⾔࠹ᾦమᇰ㣬࡛ࠉ⡷ࡷ㯇࡝࡜ࡡᅖమ୕࡚ᇰ㣬ࢅ⾔࠹ᅖమᇰ㣬ࡡ 2 ⛸㢦࠿Ꮛᅹࡌࡾࠊ㔂㏸Ⓠ㓕⏐ᴏ࡚㔔さ࡝㓕⣪ࡢࠉᅖమᇰ㣬࡚ࡊ࠾⏍⏐ࡈࡿ࡝࠷∸࠿ኣࡂᅖమᇰ㣬࡚ࡡよᯊ࠿㟸ᖏ࡞㔔さ࡚࠵ࡾࠊࡊ࠾ࡊ࡝࠿ࡼࠉ
ᅖమᇰ㣬࡚ࡢᇰᆀ࡛Ⳟమࢅฦ㞫ࡌࡾࡆ࡛࠿ᅏ㞬࡚ࠉᇰ㣬㏭୯࡞ᇰᆀ᮪௲ࢅን໩ࡈࡎࡾࡆ࡛࠿㞬ࡊ࠷ࠊࡐࡡࡒ
ࡴᇰᆀᠺฦࡡን໩࡞ࡻࡾࠉ㐿ఎᏄⓆ⌟ࡡよᯊ࠿୘ྊ⬗࡚࠵ࡖࡒࠊࡐࡆ࡚௑ᅂࠉᅖమᇰ㣬࡛ྜྷᵕ࡝᮪௲࡚ᇰ㣬࠿࡚ࡀࠉᇰᆀᠺฦ࠿ฦ㞫ྊ⬗࡝ࣆ࢔ࣜࢰ࣭ᇰ㣬Ἢ 1)࡚㯔Ⳟࢅ⏍⫩ࡈࡎࠉDNA ࢛ࣛࢥ࣏࢕ࢠࣞ࢓ࣝ࢕ࢅ⏕࠷࡙㐿ఎᏄࡡ⥑⨮ⓏⓆ⌟よᯊࢅ⾔࡝ࡖࡒࡡ࡚ሒ࿈ࡌࡾࠊ
<᪁Ἢ࠽ࡻࡦ⤎ᯕ> ௑ᅂࡡⓆ⌟よᯊ࡞ࡢࠉ㯔Ⳟࡡࢣࢿ࣑よᯊ࡚᪺ࡼ࠾࡞࡝ࡖࡒ࡮࡯ධ࡙ࡡ㐿ఎᏄ࡞ᑊᚺࡌࡾࠉ
11000 ࡡ≁␏Ⓩࣈ࣭ࣞࣇࢅࢪ࣎ࢴࢹࡊࡒ࢛ࣛࢥ࢓ࣝ࢕ࢅ౐⏕ࡊࡒ 2)ࠊࣆ࢔ࣜࢰ࣭ᇰ㣬࡚㯔Ⳟࢅᇰ㣬ࡊࡒᚃࠉRNA ࢅ᢫ฝࡊ࡙࢓ࣝ࢕⏕ࣈ࣭ࣞࣇࢅష⿿ࡊࠉ㐿ఎᏄࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣜࡡよᯊࢅ⾔࡝ࡖࡒࠊ1) H. ISHIDA et al. J. Ferment. Bioeng., 86: 301-307 (1998)2) ఫ㔕ࡼ 㸯᪝ᮇ⏍∸ᕝᏕኬఌさ᪠ࠉ P138 (2004)
Gene expression analysis under filter culture condition of Aspergillus oryzae
Motoaki Sano, Akiko Kobayashi, Masayuki Machida*, Shinichi Ohashi(KIST, *AIST)

O-2ࢷ࣒ࣞ࢓௛ຊ࣊ࢠࢰ࣭࡞ࡻࡾ㯔Ⳟᙟ㈻㌹ᥦ࡛ᑙථ )3& ࡡよᯊᴃᮇ៿ୌࠉཿᕖ⫩௥ࠉᮄᮟኣỜࠉᮿୖ┷⏜⨶ࠉ㕝ᮄ⪵ࠉ᯵ᮄ㇇ࠉ㸝㣏⥪◂㸞࠘┘Ⓩ࠙ⁿ⩽ࡼࡢ㯔Ⳟ Aspergillus oryzae ࡡ᯹Ⰵమᮆ❻࡞న⨠ࡌࡾࢷ࣒ࣞ࢓ࢅฦ㞫ࡊࠉࡐࡡᵋ㏸࠿ 12bp
㸝TTAGGGTCAACA㸞ࢅ༟న࡛ࡊࡒ⧖ࡽ㏁ࡊ㒼า࡚࠵ࡾࡆ࡛ࢅよ᪺ࡊࡒ㸦㸞ࠊ㓕ẍ➴ࡡࢷ࣒ࣞ࢓ࡢ᯹Ⰵమࡡ々⿿ࡷᏭᏽ໩࡞㛭ࢂࡾࡆ࡛࠿▩ࡼࡿࡾ࠿ࠉ㯔Ⳟࢷ࣒ࣞ࢓࡞ࡗ࠷࡙ࡢࡐࡡ々⿿ᶭᵋࡷ᯹Ⰵమᮆ❻ಕᣚᶭᵋ➴ࡡᇱ♇Ⓩ▩ぜ࠿よ᪺ࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚㯔Ⳟࢷ࣒ࣞ࢓ࢅ௛ຊࡊࡒ࣊ࢠࢰ࣭ࢅషᠺࡊࠉ⣵⬂ᑙථᚃࡡ≁ᛮࢅ᳠ゞࡊࡒࠊ࠘᪁Ἢཀྵࡦ⤎ᯕ࠙ࢷ࣒ࣞ࢓㒼า㸝135bp㸞ࢅ㸦ࢤࣅ࣭ࠉ࠵ࡾ࠷ࡢ㏣ྡྷࡀ࡞ 2 ࢤࣅ࣭᭯ࡊࠉ࣏࣭࣭࢜㐿ఎᏄ ptrA
ࢅ㏻⤎ࡊࡒࣈࣚࢪ࣐ࢺ࣊ࢠࢰ࣭ࢅష⿿ࡊࡒࠊ࣊ࢠࢰ࣭୕ࡡ ptrA ࡢ MfeI ヾㆉ㒼าࢅ୕Ὦഁ࡞᭯ࡊࠉୖὮ࡞ࢷࣞ
࣒࢓㒼าࢅ㏻⤎ࡌࡾࡻ࠹㒼⨠ࡊࡒࠊࡆࡿࡼࡡ࣊ࢠࢰ࣭ࢅ⏕࠷࡙ A. oryzae NFRI1599 ᰬࡡᙟ㈻㌹ᥦࢅ⾔ࡖࡒࠊᚋࡼࡿࡒᙟ㈻㌹ᥦమࡡࢣࢿ࣑ DNA ࢅ MfeI ࡚ᾐ໩ࡊࠉࢦࢧࣤよᯊࢅ⾔ࡖࡒ⤎ᯕࠉᙟ㈻㌹ᥦᰬ࡞≁᭯࡝ ptrA ࢨࢡࢻࣜ࠿ 2-6 ᮇ᳠ฝࡈࡿࠉ࣊ࢠࢰ࣭ࡢኣࢤࣅ࣭࡚ᑙථࡈࡿ࡙࠷ࡒࠊࡆࡿࡼࡡࢨࢡࢻࣜࡢᐙ୹ཀྵࡦᙟ㈻㌹ᥦᰬ
࡞භ㏳࡝ࢷ࣒ࣞ࢓ࢨࢡࢻࣜ㸝10-12 ᮇࠉⳞᰬ࡞ࡻࡽ␏࡝ࡾ㸞ࡡୌ㒂࡛⛛ິᗐ࠿ୌ⮬ࡊࡒࠊᙟ㈻㌹ᥦᰬ࡞ࡻࡖ࡙
ࡢࠉᐙ୹ᰬ࡞࡝࠷᩺ࡒ࡞⏍ࡋࡒ ptrA ࢨࢡࢻ࡛ࣜࠉྜྷᵕ࡞᩺ࡒ࡞⏍ࡋࡒࢷ࣒ࣞ࢓ࢨࢡࢻࣜࡡ⛛ິᗐ࠿ୌ⮬ࡌࡾ
ᰬࡵぜࡼࡿࡒ㸝᳠ฝ⠂ᅑ⣑ 1 – 15kb㸞ࠊ௧୕ࡡ⤎ᯕ࠾ࡼࠉᑙථ࣊ࢠࢰ࣭ࡢ々ᩐࡡࢷ࣒ࣞ࢓ࡡ㎾എ࡞⤄ࡲ㎲ࡱࡿࠉ࣊ࢠࢰ࣭⏜ᮮࡡࢷ࣒ࣞ࢓࠿ ptrA ࢅ㏻⤎ࡊࡒࡱࡱ᯹Ⰵమᮆ❻࡛ࡊ࡙ᶭ⬗ࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
1) ᴃᮇࡼࠉ᪝ᮇ㎨ⰹ໩Ꮥఌ 2003 ᖳᗐኬఌㅦⁿさ᪠㞗 p29㸝2003㸞
Transformation of Aspergillus oryzae with the telomere-attached vectors and characterization of the introducedDNA
Ken-Ichi Kusumoto, Ikuyo Furukawa, Tae Kimura, Mayumi Matsushita, Satoshi Suzuki, Yutaka Kashiwagi(Natl. Food Res. Inst.)

41

O-3
⣊≟Ⳟ࣬㓕ẍ࡞࠽ࡄࡾ RNA ࢦ࢕ࣝࣤࢨࣤࢡ⤊㊪ࡡ㐅໩Ⓩよᯊ୯ᒁᩔᆍ㸡みㆺ├ᶖ㸡Nguyen Bao Quoc㸡┶ᒜ⁘ᚷ㸝♼ᡖኬ࣬㎨Ꮥ㒂㸞RNA ࢦ࢕ࣝࣤࢨࣤࢡࡢ dsRNA ࢅ௒ࡊࡒ㌹෕ᚃࡡ㐿ఎᏄᢒโᶭᵋ࡚࠵ࡾ࠿ࠉࡆࡡ⤊㊪࡞ࡢ RNaseIII ᵕࢽࢠࣝ࢓࣭ࢭ࡚࠵ࡾ DicerࠉRNA ౪Ꮛ RNA ࣒࣭࣎ࣛࣚࢭ(RdRP)ࠉRNA induced silencing complex (RISC)ࡡᵋᠺさ⣪࡚࠵ࡾ Argonaute ࡝࡜ࡡ⺦Ⓣ㈻࠿㛭୙ࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ࡙࠷ࡾࠊࢣࢿ࣑㒼า᝗ሒ࠿ࠉප㛜ࡈࡿ࡙࠷ࡾ㸦㸥⛸ࡡᏄࡡ࠹Ⳟࠉ㸩⛸ࡡᢰᏄⳞࠉ㸦⛸ࡡ᥃ྙⳞ࡞࠽ࡄࡾࡆࡿࡼࡡ⺦Ⓣ㈻㒼าࢅㄢᰕࡊࡒᡜࠉ㸨⛸ࡡᏄࡡ࠹Ⳟ(Candida albicans, C. lusitaniae, Saccharomyces cerevisiae)ࠉ㸦⛸ࡡᢰᏄⳞ(Ustilago maydis)࡞࠽࠷࡙ࠉ୕オࡡRNA ࢦ࢕ࣝࣤࢨࣤࢡ⺦Ⓣ㈻ࡡ࣓࣌ࣞࢡࡡࡌ࡬࡙࠵ࡾ࠷ࡢࡐࡡ࡮࡛ࢆ࡜࠿Ꮛᅹࡊ࡙࠽ࡼࡍࠉRNA ࢦ࢕ࣝࣤࢨ
ࣤࢡ⤊㊪࠿኶ࢂࡿ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊୌ᪁ࠉᑊ↯Ⓩ࡞ᢰᏄⳞ࡚࠵ࡾ Coprinus cinereus ࡚ࡢࠉ㸬⛸ࡡ
Argonauteࠉ㸨⛸ࡡ Dicer, 㸭⛸ࡡ RdRP ࣂࣚࣞࢡࢅࢣࢿ࣑හ࡞ಕ᭯ࡊ࡙࠽ࡽࠉᴗࡴ࡙々㞟࡝ RNA ࢦ࢕ࣝࣤࢨ
ࣤࢡ⤊㊪ࢅಕ᭯ࡊ࡙࠷ࡾࡆ࡛࠿᥆ᏽࡈࡿࡒࠊࡆࡿࡼࡡ⤎ᯕࡢࠉ⣊≟Ⳟ࡞࠽࠷࡙ RNA ࢦ࢕ࣝࣤࢨࣤࢡ⤊㊪ࡢࠉࡐࡡⳞࡡ⏍ឺࡷᙟឺฦ໩ࡡ⛤ᗐ࡞ᚺࡋ࡙ኣᵕᛮ࡞ᐣࢆࡓฦ໩ࢅ㐑ࡅ࡙࠷ࡾࡆ࡛ࢅ♟ြࡊࡒࠊ྘Ⳟ࡞࠽ࡄࡾ
RNA ࢦ࢕ࣝࣤࢨࣤࢡ⺦Ⓣ㈻ࡡฦᏄ⣌⤣よᯊࢅ⾔ࡖࡒ⤎ᯕࠉ⮾࿝῕࠷ࡆ࡛࡞ୌࡗࡡⳞහࡡࣂࣚࣞࢡྜྷኃࡻࡽࡵࠉ
Ꮔࡡ࠹Ⳟ࣬ᢰᏄⳞ࣬᥃ྙⳞ࡞࠽ࡄࡾ࢛࣭ࢮࣞࢡࡡ᪁࠿ࡻࡽ㎾⦍࡚࠵ࡾౚ࠿♟ࡈࡿࡒࠊࡆࡡ⤎ᯕࡢࠉ々ᩐࡡ RNAࢦ࢕ࣝࣤࢨࣤࢡ⤊㊪ࡡ㉫″࠿ኬንཿࡂࠉ୹さ࡝Ⳟ⩄ࡡฦ໩௧๑࡞Ꮛᅹࡊ࡙࠷ࡒࡆ࡛ࢅ♟ြࡌࡾࡵࡡ࡚࠵ࡖࡒࠊ
Ancient origin of RNA silencing pathways in fungi
Hitoshi Nakayashiki, Naoki Kadotani, Nguyen Bao Quoc, Shigeyuki Mayama(Fac. of Agri., Kobe Univ.)

O-4
EGFP ࡛ AoVam3p ࡡ⼝ྙࢰࣤࣂࢠ㈻ࢅ⏕ ࠷ࡒᾦ⬂⭯ࡡྊっ໩࡞ࡻࡾ㯔Ⳟ A. oryzae ᾦ⬂ࡡᙟឺよᯊ
ḿ㊪ῗஒ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞࠘┘Ⓩ࠙⣊≟Ⳟ࡞࠽࠷࡙ᾦ⬂ࡢࠉࢲ࣭ࣖࣇ≟ᾦ⬂ࡡᏋᅹ࡝࡜ฝⰾ㓕ẍ࡚ࡢぜࡼࡿ࡝࠷≁ᚡࢅങ࠻࡙࠷ࡾ㸣ᠻࠍࡢࡆࡿࡱ࡚ࠉ㯔Ⳟ A. oryzae ࡞࠽࠷࡙ EGFP ࡛ᾦ⬂⭯୕ࡡ t-SNARE ࡛ࡊ࡙ᶭ⬗ࡌࡾ࡛஢᝷ࡈࡿࡾ AoVam3p
ࡡ⼝ྙࢰࣤࣂࢠ㈻(EGFP-AoVam3p)ࢅ⏕࠷ࡾࡆ࡛࡞ࡻࡽࠉᾦ⬂ິឺࡡよᯊࢅ⾔ࡖ࡙ࡀࡒ 1) 㸣௑ᅂᠻࠍࡢ
EGFP-AoVam3p ࢅ⏕࠷ࡒᾦ⬂ふᐳ⣌ࢅࡈࡼ࡞Ⓠᒈࡈࡎࠉฦ⏍ᏄⓆⰾࠉẴ୯Ⳟ⣊ࠉᇱᗇ㒂ࡡཿ࠷Ⳟ⣊࡛࠷ࡖࡒ
ᵕࠍ࡝Ⓠ㐡ṹ㝭ࡡⳞ⣊࡞࠽ࡄࡾᾦ⬂ᙟឺࢅふᐳࡊࡒ㸣࠘⤎ᯕ࠙⣵⬂᯹Ⰵ࡛ࡡẒ㍉࠾ࡼࠉEGFP-AoVam3p ࡢᾦ⬂࡞ຊ࠻ࠉฝⰾ㓕ẍࡡᚃ᭿࢙ࣤࢺࢮ࣭࣑࡞㢦జࡊࡒࢤࣤࣂ࣭ࢹ࣒ࣤࢹ࡞ࡵᑻᅹࡌࡾࡆ࡛࠿ฦ࠾ࡖࡒ㸣ࡱࡒࠉฦ⏍ᏄⓆⰾ᫤ࡡᾦ⬂ᙟឺࢅふᐳࡊࡒ࡛ࡆࢀࠉฦ⏍Ꮔࡡ⭶₮࡞ఔࡖ࡙ᾦ⬂࠿ኬࡀࡂⓆ㐡ࡊ࡙࠷ࡂࡆ࡛࠿ふᐳࡈࡿࡒ㸣ࡈࡼ࡞ᇰᆀ࡛├᥃᥃ࡊ࡙࠷࡝࠷Ẵ୯Ⳟ⣊࡞࠽࠷࡙ࡢࠉኬࡀࡂⓆ㐡ࡊࡒᾦ⬂ࡢ㏳ᖏふᐳࡈࡿࡍࠉ࠾ࢂࡖ࡙ࢲ࣭ࣖࣇ≟ᾦ⬂࡛ᑚࡈ࡝ᾦ⬂࡞ࡻࡾࢾࢴࢹ࣭࣠ࢠ
ᵋ㏸࠿ふᐳࡈࡿࡒ㸣ࡱࡒᇱᗇ㒂ࡡཿ࠷Ⳟ⣊࡞࠽࠷࡙ࡲࡼࡿࡾⓆ㐡ࡊࡒᾦ⬂࡚ࡢ EGFP ⺧ක࠿ᾦ⬂හ⭅࡞ふᐳࡈࡿࡒࡆ࡛࠾ࡼࠉࡆࡿࡼࡡⳞ⣊࡞࠽࠷࡙ࡢ࢛࣭ࢹࣆ࢒ࢩ࣭࡞ࡻࡾ⣵⬂㈻ᠺฦࡡฦよ࡛ࠉࡐࡿ࡞ఔ࠹ᾦ⬂⭯ࡡཱི㎲ࡲ࠿࠽ࡆࡖ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾ㸣௧୕ࡡ⤎ᯕ࠾ࡼࠉ⣊≟Ⳟࡡᾦ⬂ࡢᵕࠍ࡝Ⓠ㐡ṹ㝭࡞࠽࠷࡙≁␏Ⓩ࡝഼
ࡀࢅᢰࡖ࡙࠷ࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒ㸣1)ḿ㊪ࡼࠉ➠㸨ᅂ⣊≟ⳞฦᏄ⏍∸Ꮥࢤࣤࣆ࢒ࣝࣤࢪさ᪠㞗㸡p48
Detailed observations of vacuoles using a fusion protein of EGFP with AoVam3p in A. oryzae.
Jun-ya SHOJI, Manabu ARIOKA㸡Katsuhiko KITAMOTO (Dept. of Biotech., Univ. of Tokyo)

42

O-5
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪ⤊㊪ࡡよᯊᵵཾ⿩ḗ㑳ࠉ୯℀ᬓ஄ࠉḿ㊪ῗஒࠉ᭯ᒱ Ꮥࠉ໪ᮇົࡥࡆ (᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ)
࠘┘Ⓩ࢙࠙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪࡡᶭᵋࡢࠉິ∸⣵⬂࠽ࡻࡦ㓕ẍ࡞࠽࠷࡙◂✪࠿㐅ࢆ࡚࠷ࡾࡵࡡࡡࠉ⣊≟Ⳟ࡞࠽࠷࡙ࡢ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪ⮤మ࠿Ꮛᅹࡌࡾ࠾ྫྷ࠾࡚ࡆࡿࡱ࡚㆗ㄵ࠿࡝ࡈࡿ࡙ࡀࡒࠊ᭩㎾ A. nidulans ࡞࠽࠷࡙ࠉ
ࣈࣛࣤࢹࣚࣤࢪ࣭࣎ࢰ࣭࡚࠵ࡾ UapC ࢰࣤࣂࢠ㈻࠿ᇰᆀ࡫ࡡ࢓࣓ࣤࢼ࣑ࢗሲࡡ῟ຊ࡞ࡻࡖ࡙⣵⬂⭯࠾ࡼᾦ⬂࡫࡛㍲㏞ࡈࡿࡾࡆ࡛࠿ሒ࿈ࡈࡿࠉ⣊≟Ⳟ࡞࠽࠷࡙ࡵ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪࡡᏋᅹ࠿ᙁࡂ♟ြࡈࡿࡒࠊࡐࡆ࡚ᮇ
◂✪࡚ࡢ⣊≟Ⳟ࡞࠽ࡄࡾ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪᶭᵋࡡよᯊࢅ┘Ⓩ࡛ࡊ࡙ A. oryzae ࡞࠽ࡄࡾྜྷ⤊㊪ࡡྊっ໩ࢅ⾔ࡖࡒࠊ࠘᪁Ἢ࡛⤎ᯕ࠙A. oryzae ࡞࠽ࡄࡾ uapC ┞ྜྷ㐿ఎᏄ AouapC ࢅࢠ࣭ࣞࢼࣤࢡࡊࠉࡐࡡ⏐∸ࡡ C ᮆ❻࡞ EGFP ࢅ
⼝ྙࡊࡒ AoUapC-EGFP ࢅ A. oryzae NS4 ᰬ࡞࠽࠷࡙ amyB promoter ࡡโᚒୖ࡚Ⓠ⌟ࡈࡎࡾᰬࢅష⿿ࡊࡒࠊභ
↌Ⅴ࣭ࣝࢧ࣭㢟᚜㙶ふᐳ࡞ࡻࡽ AoUapC-EGFP ࡢ⣵⬂⭯࡞ᑻᅹࡊ࡙࠽ࡽࠉ࢓࣓ࣤࢼ࣑ࢗሲࢅྱࡳᇰᆀ࡞ࢨࣆ
ࢹࡌࡾ࡛ࠉ⣵⬂⭯ࡡ EGFP ⺧ක࠿⷟ࡿ࡙࠷ࡂᵕᏄ࠿ふᐳࡈࡿࡒࠊࡱࡒࡆࡡཱིࡽ㎲ࡲ࠿ୌ⯙Ⓩ࡝࢙ࣤࢺࢦ࢕ࢹ
࣭ࢨࢪࡡ≁ᚡࢅങ࠻࡙࠷ࡾࡆ࡛ࢅ ATP ྙᠺ㜴ᐐ๠ࡷ࢓ࢠࢲࣤ⬲㔔ྙ๠➴ࢅ⏕࠷࡙☔ヾࡊࡒࠊ௧୕ࡡࡆ࡛࠾ࡼ
AoUapC-EGFP ࠿࢓࣓ࣤࢼ࣑ࢗሲࡡᏋᅹୖࠉ⣵⬂⭯࠾ࡼ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪ࡞ࡻࡽཱིࡽ㎲ࡱࡿࡾࡆ࡛ࢅ♟ࡊࠉ⣊≟Ⳟ࡞࠽ࡄࡾ࢙ࣤࢺࢦ࢕ࢹ࣭ࢨࢪࡡᏋᅹࢅิࡴ࡙☔ヾࡊࡒࠊࡈࡼ࡞ࠉྜྷ⤊㊪ふᐳࡡ㝷࡞ࠉᾦ⬂᯹Ⰵモ࡚ⷾ࠵ࡾ CMAC ࡞ࡻࡖ࡙᯹Ⰵࡈࡿ࡝࠷㢓⢇≟ࡡ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మ࠿ぜฝࡈࡿࡒࠊ᚜ᑚ⟮⬲㔔ྙ๠࡞ࡻࡽࡆࡿ
ࡼࡡᵋ㏸మࡡິࡀ࠿ೳḾࡊࡒࡆ࡛࠾ࡼࠉ࢙ࣤࢺࢮ࣭࣑ᵕᵋ㏸మࡢⳞ⣊හࢅ᚜ᑚ⟮࡞Ἒࡖ࡙⛛ິࡊ࡙࠷ࡾࡆ࡛
࠿ࢂ࠾ࡖࡒࠊ
Analysis of endocytic pathway in Aspergillus oryzae
Yujiro Higuchi, Tomoyuki Nakahama, Jun-ya Shoji, Manabu Arioka, Katsuhiko Kitamoto (Dept. of Biotech., Univ. ofTokyo)

O-6
㯔Ⳟ(Aspergillus oryzae)ࡡ impala ᵕ DNA ࢹࣚࣤࢪ࣎ࢯࣤ Aoimp1 ࡡⳞᰬ㛣ኣᵕᛮ࡛㌹⛛Ὡᛮᑚ➗ཋ༡ಘ㸦㸡㸧㸡ᑚ⏷ ᾀ㸨㸡⛑ Ὂ஦㸨㸡㧏ᶣ▹⧂㸧㸡஫࿝ົஒ㸦㸝㸦᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ㸡㸧⚽⏛┬⥪㣏◂࣬⏍∸ᶭ⬗㸡㸨᭮᰿෗࣬⥪◂㸞

࠘┘Ⓩ࠙ᐁ⏕㯔Ⳟ Aspergillus oryzae OSI1013 ᰬ࠾ࡼぜฝࡈࡿࡒ᩺ぞࢹࣚࣤࢪ࣎ࢯࣤ Aoimp1 ࡢ,㌹⛛Ὡᛮ࠿㧏࠷ Fusarium ᒌࡡ impala㸝Tc1/mariner superfamily㸞࡛㧏࠷┞ྜྷᛮࢅ᭯ࡊ,㯔Ⳟ࡞࠽ࡄࡾ㌹⛛Ὡᛮ࠿᭿ᙽࡈࡿࡒ㸣ࡐࡆ࡚,ࢹࣚࣤࢪ࣎ࢯࣤࢅ⏕࠷ࡒඁⰃᰬࡡ⫩⛸ࡷ㐿ఎᏄ Tagging Ἢࡡ㛜Ⓠࢅ┘Ⓩ࡛ࡊ࡙,Ⳟᰬ㛣࡞࠽ࡄࡾࢹࣚࣤ
ࢪ࣎ࢭ࣭ࢪ㸝aotA㸞㒼าࡡฦᕱ࡛ᵋ㏸ࢅよᯊࡌࡾ࡛ྜྷ᫤࡞㌹⛛Ὡᛮ࡞ࡗ࠷࡙ࡵ᳠ゞࡊࡒ㸣࠘᪁Ἢ࡛⤎ᯕ࠙ITR 㒂ฦࢅࣈࣚ࢕࣏࣭࡞⏕࠷ࡒ PCR ࡞ࡻࡽ,RIB40 ᰬࢅྱࡳ A.oryzae ࡷ A.sojae ౩モᰬࡡኣࡂ࡞ᗀࡂฦᕱ࠿ヾࡴࡼࡿ,ࢦࢧࣤよᯊ࡞ࡻࡾࢤࣅ࣭ᩐࡡ㐢࠷࠾ࡼ㯔Ⳟ౩モᰬࡢ 3 ࢡ࣭ࣜࣈ࡞ฦ㢦ࡈࡿࡒ.aotA ࡡሲᇱ㒼าࢅỬᏽࡊࡒ࡛ࡆࢀ,OSI1013 ➴ࡡኣࢤࣅ࣭ᰬ࡚ࡢ࢓࣐ࢿ㓗㒼า࠿ୌ⮬ࡊ࡙࠷ࡾࡡ࡞ᑊࡊ,RIB209 ᰬ㸝3ࢤࣅ࣭㸞࡚ࡢ 2 ಴ࡡ࢓࣐ࢿ㓗ን␏,RIB40 ➴ࡡ 1 ࢤࣅ࣭ᰬ࡚ࡢ㧏㢎ᗐࡡ RIP ᵕ㸝GCЍAT㸞ን␏࠿ヾࡴࡼࡿࡒ㸣ୌ᪁㸡OSI1013 ᰬࡡࢿ࣭ࢧࣤよᯊ࡞࠽࠷࡙ CuSO㸩῟ຊᇰ㣬ࡷ㧏Ὼᇰ㣬᫤࡞ aotA ࡡ㌹෕㔖୕᪴࠿ヾࡴࡼࡿࡒࡡ
࡚㸡ྜྷᵕ᮪௲ୖ࡚ᇰ㣬ࡊࡒฦ⏍Ꮔ࠾ࡼࡡ Transposon-Trapping ࢅ⾔ࡖࡒ㸣ࡐࡡ⤎ᯕ㸡niaD ࡡࡲ࡝ࡼࡍ crnA 㐿ఎᏄ࡫ࡡ᣼ථࡵヾࡴࡼࡿ㸡Aoimp1 ࡢࢪࢹࣝࢪ᮪௲ୖ࡚㌹⛛Ὡᛮࢅ♟ࡌࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒ㸣
Distribution and transposition of impala-like DNA transposon Aoimp1 in Aspergillus oryzae
Hironobu Ogasawara1,㸧, Hiroshi Obata㸨, Yoji Hata㸨, Saori Takahashi㸧, Katsuya Gomi㸦
(㸦Akita Res. Inst. Food and Brewing, Bioeng., 㸧Div. Biosci. Biotech. Future Bioind., Grad. Sch. Agric. Sci., Tohoku
Univ.㸡㸨Res.Inst.,Gekkeikan Sake Co.)

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ࢠ ࢙ ࣤ 㓗 ⏍ ⏐ ⣊ ≟ Ⳟ Aspergillus niger ࡞ ࠽ ࡄ ࡾ Ὡ ᛮ 㓗 ⣪ ⛸ ࡞ ᑊ ࡌ ࡾ ⪇ ᛮ ࡫ ࡡ
alternative oxidase ࡡᐞ୙᭱㒂㈏⃀ࠉᮄ㔕㑝ჹࠉ᱊ᮟකኯ㑳 㸝᪡ኬ࣬⌦ᕝ࣬ᚺ໩㸞࠘┘Ⓩ࠙ࢠ࢙ࣤ㓗⏍⏐⣊≟Ⳟ A. niger WU-2223L ࡢ㏳ᖏࡡࢨࢹࢠ࣑ࣞ࿣ྺ⣌ࡡ௙࡞ࢨ࢓ࣤ㟸វུᛮ࿣ྺ⣌ࢅ᭯ࡌࡾࠊⁿ⩽ࡼࡢࠉᙔル࿣ྺ⣌ࢅᵋᠺࡌࡾ㓕⣪ alternative oxidase ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ(aox1)ࢅཱིᚋࡊ࡙࠷ࡾ1)ࠊ
ᮇ◂✪࡚ࡢ㹹Ὡᛮ㓗⣪⛸࡞ᑊࡌࡾ⪇ᛮ࡫ࡡ alternative oxidase ࡡᐞ୙ࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙ࠉaox1◒ቪᰬ(DAOX-1)࡞ࡗ࠷࡙Ὡᛮ㓗⣪⛸ࢅྱࡳᇰᆀ࡚ࡡ⏍Ꮛ⋙ࢅㄢ࡬ࡒࠊࡈࡼ࡞ࠉDAOX-1 ࡞࠽ࡄࡾ catalase(CAT)Ὡᛮ࡛ superoxide dismutase(SOD)ὩᛮࢅῼᏽࡊࠉὩᛮ㓗⣪⛸ࡡ㝎ཡ࡞㛭୙ࡌࡾ㓕⣪ࡡ⏍⏐࡞ aox1 ◒ቪࡡཀྵ࡯ࡌᙫ㡢࡞ࡗ࠷࡙᳠ゞࡊࡒࠊ࠘᪁Ἢ࠽ࡻࡦ⤎ᯕ࠙ A. niger WU-2223L ⏜ᮮࡡ aox1 ◒ቪᰬ(DAOX-1)ࡢ㹹┞ྜྷ⤄ᥦ࠻࡞ࡻࡖ࡙షᠺࡊࡒࠊDAOX-1 ࡚ࡢ aox1 ࡡ㌹෕࠽ࡻࡦ alternative oxidase Ὡᛮ࠿ᾐ኶ࡊࡒࡆ࡛ࢅ☔ヾࡊࡒࠊDAOX-1 ࡞ࡗ࠷࡙ࡢ㹹Ὡᛮ㓗⣪⛸࡛ࡊ࡙ 5 mM ࡡ㐛㓗໩Ề⣪ࢅྱࡳᇰᆀ୕࡚ࡡ⏍Ꮛ⋙࠿ WU-2223L ࡡ⏍Ꮛ⋙ࡡ⣑ 20%࡛࡝ࡽࠉࡈࡼ࡞
t-butyl hydroperoxide ࢅྱࡳᇰᆀ࡚ࡡ⏍Ꮛ⋙ࡵ఩ୖࡊࡒࡆ࡛࠾ࡼὩᛮ㓗⣪⛸࡞ᑊࡌࡾ⪇ᛮࡡ఩ୖ࠿ヾࡴࡼࡿࡒࠊ
5 mM ࡡ㐛㓗໩Ề⣪ࢅྱࡳᇰᆀ࡚ 4 ᪝㛣⏍⫩ࡈࡎࡒ DAOX-1 ࡚ࡢࠉWU-2223L ࡛Ẓ㍉ࡊ࡙ CAT ẒὩᛮ࠿⣑ 1.2
ಶࠉSOD ẒὩᛮ࠿ 10 ಶ௧୕࡞ቌኬࡊࡒࠊ௧୕ࡡ⤎ᯕࡢࠉaox1 ◒ቪ࡞࡛ࡵ࡝࠹ alternative oxidase Ὡᛮࡡᾐ኶
࠿ DAOX-1 ࡡ⣵⬂හ࡞࠽࠷࡙Ὡᛮ㓗⣪⛸ࢅቌኬࡈࡎࠉSOD Ὡᛮ࠿ቌኬࡌࡾࡆ࡛ࢅᙁࡂ♟ြࡊࡒࠊ1) K. Kirimura et al., Curr. Genet., 34, 472-477 (1999)
Contribution of Alternative Oxidase to Resistance to Reactive Oxygen Species in Aspergillus niger Producing
Citric Acid
Takasumi Hattori, Kuniki Kino, Kohtaro Kirimura(Dept. Appl. Chem., Sch. Sci. Eng., Waseda Univ.)

O-8࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ APSES ࢰࣤࣂࢠ㈻ Mstu1 ࡢ௛╌ჹࡡᠺ⇅ࢅโᚒࡌࡾ
こᮟ㯖㔓Ờ㸡ᯐ㛏⏍ 㸝⏍∸◂㸞⣊≟Ⳟ࡞࠽࠷࡙ࠉAPSES ㌹෕ᅄᏄࡢⳞࡡᙟឺฦ໩ࢅโᚒࡌࡾࠊ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ APSES ㌹෕ᅄᏄ Mstu1 ࡡḖ᥾ᰬࡢࠉⳞ⣊⏍㛏ࡡ㐔ᘇࠉ⬂Ꮔᙟᠺᩐࡡ఩ୖ㸝㔕⏍ᆵࡡ⣑ 10%㸞ࠉஹ㒼᫤ࡡ㞜ᛮḖ᥾ࠉᄂ㟕᥃⛸᫤ࡡវ᯹⋙ࡡ఩ୖࢅ♟ࡌࠊMstu1 ࡞࠽ࡄࡾ௛╌ჹ㸝វ᯹≁␏Ⓩჹᏻ㸞ᙟᠺࢅふᐳࡊࡒ࡛ࡆࢀࠉḖ᥾ᰬ࡚ࡢิ᭿ࡡ௛╌ჹᙟᠺ㸝<8h㸞࡞㐔ᘇ࠿ࡲࡼࡿࡒࡵࡡࡡࠉ16 ᫤㛣ᚃ࡞ࡢ㔕⏍ᆵᰬ࡛ྜྷ⛤ᗐࡡᩐࡡࠉᙟឺⓏ࡞ḿᖏ࡝௛╌ჹࢅᙟᠺࡊࡒࠊࡊ࠾ࡊ࡝࠿ࡼࠉ౴ථⳞ⣊ࡡᙟᠺ⋙ࡢ Mstu1 Ḗ᥾ᰬ࡚ࡢ㔕⏍ᆵᰬࡡ⣑ 20%ࡱ࡚఩ୖࡊ࡙࠷ࡒࠊࡱࡒࠉ
௛╌ჹᙟᠺㄇᑙ 24 ᫤㛣ᚃࡡ Mstu1 Ḗ᥾ᰬ࡚ࡢⓆⰾ⟮ࡡ␏ᖏ࡝ฦᒪ࠿ふᐳࡈࡿࡒࠊ࠷ࡵࡔ⑋Ⳟ࡚ࡢࠉ⬂Ꮔ/ฦ
⏍Ꮔᯮᙟᠺࠉ௛╌ჹᙟᠺࠉ౴ථⳞ⣊ᙟᠺ࠿ cAMP ࢨࢡࢻࣜࡡโᚒࢅུࡄ࡙࠷ࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾ࠿ࠉMstu1
Ḗ᥾ᰬࡡ࠷ࡍࡿࡡḖ᥾ᙟ㈻࡞ࡗ࠷࡙ࡵ cAMP ῟ຊ࡞ࡻࡽⴥࡊ࠷ᅂᚗࡢぜࡼࡿ࡝࠾ࡖࡒࠊMstu1 Ḗ᥾ᰬ⬂Ꮔࢅ↋ഭࡡ࢕ࢾ࡫ᄂ㟕᥃⛸ࡊࡒሔྙ࡞ࡢ⑋ᩤ࠿࡮࡛ࢆ࡜ぜࡼࡿ࡝࠷࠿ࠉഭࢅࡗࡄࡒ࢕ࢾ࡞᥃⛸ࡊࡒሔྙ࡞ࡢ࢕ࢾ
࡞౴ථࡊ⑋ᩤࢅᙟᠺࡊࡒࠊ࠷ࡵࡔ⑋Ⳟࡢ௛╌ჹහ࡞㧏⃨ᗐ࡞⵫✒ࡈࡿࡒࢡࣛࢬ࣭ࣞࣜ࡞ࡻࡽᙟᠺࡈࡿࡒ⭶ᅸࢅฺ⏕ࡊ࡙࢕ࢾ࡫౴ථࡌࡾ࠿ࠉࢡࣛࢬ࣭ࣞࣜࡢ⬂Ꮔහࡡࢡࣛࢤ࣭ࢣࣤࡷ⬙㈻࠾ࡼ⏍ྙᠺࡈࡿࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊ௛╌ჹᙟᠺㄇᑙ 24 ᫤㛣ᚃ࡞࠽࠷࡙ࠉ⬂Ꮔ࣬௛╌ჹහࢡࣛࢤ࣭ࢣࣤࢅ࣭ࣙࢺ࣑࢜ࣛࢗᾦࠉ⬙㈻ࢅࢻ࢕ࣜ
ࣇ࣭࡚ࣜ᯹Ⰵࡊふᐳࡊࡒ࡛ࡆࢀࠉ㔕⏍ᆵᰬ࡛Ẓ࡬ࠉMstu1 Ḗ᥾ᰬ࡚ࡢ⬂Ꮔහࢡࣛࢤ࣭ࢣࣤ࣬⬙㈻ࡡ௛╌ჹ࡫ࡡ⛛ິ⋙࡛ฦよ⋙࠿ⴥࡊࡂ఩ୖࡊ࡙࠷ࡒࠊࡆࡿࡼࡡ⤎ᯕࡢࠉMstu1 ࠿௛╌ჹහࡡ⭶ᅸᙟᠺ࡛࠷ࡖࡒᶭ⬗Ⓩᠺ
⇅ࢅโᚒࡊ࡙࠷ࡾࡆ࡛ࢅ♟ြࡌࡾࠊ࠷ࡵࡔ⑋Ⳟࡡ cAMP-dependent protein kinase A (PKA) catalytic subunit(Cpka)ࡢ Mstu1 ࡛ྜྷᵕࠉ⬂Ꮔහࡡࢡࣛࢤ࣭ࢣࣤࡷ⬙㈻ࡡฦよ࡛௛╌ჹࡡᠺ⇅ࠉ࢕ࢾ࡫ࡡ௛╌ჹࢅ௒ࡊࡒ౴ථ
࡞㛭୙ࡌࡾࠊMstu1 ࡞ PKA ࣛࣤ㓗໩ࢦ࢕ࢹ࠿࠵ࡾࡆ࡛࠾ࡼࠉ⌟ᅹࠉCpka ࡞ࡻࡾ Mstu1 ࡡโᚒ࡞ࡗ࠷࡙᳠ゞࡊ࡙࠷ࡾࠊ
An APSES protein Mstu1 regulates appressorium maturation in Magnaporthe grisea.
Marie Nsihimura, Nagao Hayashi (NIAS)

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Cryphonectria parasitica ࡡ GTP ⤎ྙࢰࣤࣂࢠ㈻ βγ-ࢦࣇࣗࢼࢴࢹ࡛ࣆ࢚ࢪࢸ࣭ࣖࢨࣤ➗ཋ ⣸ 㸝ᐋᇖኬ࣬㣏⏐ᴏ࣬⎌ሾࢨࢪࢷ࣑㸞᪝ᮇ㸝ࡱࡒࡢ୯ᅗ㸞࠾ࡼ⡷ᅗ࡞ࡵࡒࡼࡈࡿࡒ࡛ࡈࡿࡾࢠࣛ⬏ᯜ⑋Ⳟ Cryphonectria parasitica ࡢࠉ⡷ᅗ࢓ࣂࣚࢲ
࢓ᆀ᪁࡞࠽࠷࡙ᴗࡴ࡙ඁເ࡚࠵ࡖࡒࢠࣛ(American chestnut)ࢅ࡮࡯ୌᤪࡌࡾ࡮࡜ࡡ⿍ᐐࢅࡵࡒࡼࡊࡒࠊࢗ࢕ࣜ
ࢪࢅ⏕࠷ࡒྜྷⳞࡡࣁ࢕࢛ࢤࣤࢹ࣭ࣞࣜᢇ⾙㛜Ⓠࡡୌ⎌࡛ࡊ࡙ࠉ࣒࣭ࣛࣚࣤࢺኬᏕࡡ D. Nuss ࡼࡢྜྷⳞࡻࡽࢗ
࢕ࣜࢪវ᯹ࡡᙫ㡢ࢅུࡄࡾ୔㔖మ GTP ⤎ྙࢰࣤࣂࢠ㈻ࡡ α-ࢦࣇࣗࢼࢴࢹࢅ༟㞫ࡊ࡙よᯊࢅ⾔࠷ࠉ⑋ཋᛮ࡛ࡡ
㛭ࡽࢅ♟ࡊࡒࠊࡐࡡᚃⁿ⩽ࡼࡢ β-ࢦࣇࣗࢼࢴࢹ࠽ࡻࡦ γ-ࢦࣇࣗࢼࢴࢹ㐿ఎᏄࢅ༟㞫ࡊࠉ྘ࠍࡡ⏐∸ࡡᛮ㈻ࢅ
᳠ゞࠉࡱࡒḖ᥾ን␏మࢅᚋࡒࠊβ-ࢦࣇࣗࢼࢴࢹḖ᥾ᰬࡢ≁ᚡⓏ࡝⾪⌟ᆵࢅ♟ࡊࠉPDA ᇰᆀ୕࡚ࡢ㟸ᖏ࡞㧏࠷⏍⫩⬗ࢅ♟ࡊࡒ࠿ࠉࢠࣛᖷ୕࡞࠽ࡄࡾ⑋ཋᛮࡢ࡮࡛ࢆ࡜኶ࡖ࡙࠷ࡒࠊࡱࡒྜྷ᫤᭿࡞ཱིᚋࡊࡒን␏మࡡ㸦ࡗ࠿
β-ࢦࣇࣗࢼࢴࢹḖ᥾ን␏మ࡞㓖జࡌࡾ⾪⌟ᆵࢅ♟ࡊࡒࡡ࡚よᯊࡊࡒ࡛ࡆࢀࠉን␏ࡢ β-ࢦࣇࣗࢼࢴࢹ࡚ࡵ γ-ࢦࣇࣗࢼࢴࢹࡡ㐿ఎᏄ࡚ࡵ࡝ࡂࠉβ−ࢦࣇࣗࢼࢴࢹ࡞⤎ྙࡊ࡙ࡐࡡᶭ⬗ࢅㇿ࡞ㄢ⟿ࡌࡾ࡛ࡈࡿࡾࣆ࢚ࢪࢸ࣭ࣖࢨ
ࣤᵕࢰࣤࣂࢠ㈻ࡡ㐿ఎᏄ࡞⏍ࡋ࡙࠷ࡒࠊࡆࡿࡼ β−ࢦࣇࣗࢼࢴࢹ࠽ࡻࡦࣆ࢚ࢪࢸ࣭ࣖࢨࣤᵕࢰࣤࣂࢠ㈻࡞ࡗ࠷
࡙ࠉα−ࢦࣇࣗࢼࢴࢹ࡛ࡡ┞㛭ࢅྱࡴฦᏄ࡚ࣝ࣊ࣜよᯊࡊࡒࠊࡱࡒິ∸⣵⬂ࡡࣆ࢚ࢪࢸ࣭ࣖࢨࣤ࡞ࡗ࠷࡙ᥞ၌
ࡈࡿ࡙࠷ࡒᶭ⬗࡛ࡢ␏࡝ࡾ᩺ぞࡡᛮ㈻ࢅぜฝࡊࡒࠊࡱࡒ β−ࢦࣇࣗࢼࢴࢹ࠽ࡻࡦࣆ࢚ࢪࢸ࣭ࣖࢨࣤ㐿ఎᏄ࠿ࢣࢿ࣑୕࡞࠽࠷࡙㝼᥃ࡌࡾᗑన࡞Ꮛᅹࡌࡾࡆ࡛ࠉN. crassa ࡞࠽࠷࡙ࡵྜྷᵕࡡᵋ㏸ࢅ࡛ࡾࡆ࡛ࢅぜฝࡊࡒࠊࡆࡡ
ࢨࣤࢷࢼࢴࢠ࡝ᵋ㏸ࡢ௙ࡡ㓕ẍࡷ௙ࡡ⣊≟Ⳟ࡞ࡵᗀࡂヾࡴࡼࡿࡒࠊࡱࡒ γ-ࢦࣇࣗࢼࢴࢹ࡞ࡗ࠷࡙ࡢࠉࡐࡡᵋ
㏸ࢅ N. crassa ➴࡛Ẓ㍉ࡊࡒࠊ
G-protein β γ-subunits and phosducin in the chestnut blight fungus Cryphonectria parasitica
Shin Kasahara (Dept. of Environmental Sciences, Miyagi Univ.)

O-10࣊ࢹ⑋Ⳟ, Phytophthora infestans, ࡞࠽ࡄࡾ఩Ὼ࡞ᚺ➽ࡊࡒ㐿ఎᏄⓆ⌟โᚒᶭᵋ ࡞ࡗ࠷࡙ㆺಞ἖*㸡 Howard S. Judelson 㸝Dept. of Plant Pathology, UC Riverside. * ⌟ᡜᒌ㸯㜨ᗋኬ㝌࣬⏍࿤⎌㸞ࢩࣔ࢝࢕࣓࡞វ᯹ࡊ࡙࣊ࢹ⑋ࢅᘤࡀ㉫ࡆࡌ Phytophthora infestans ࡢࠉ఩Ὼ㸝<14˚C㸞࡚ࡢ⬂Ꮔ࠾ࡼ㐗㉦Ꮔࢅᨲฝࡊࠉᬦ࠾࠷Ὼᗐ㸝>15˚C㸞࡚ࡢ⬂Ꮔ࠾ࡼ├᥃Ⳟ⣊ࢅఘ㛏ࡌࡾࠊP. infestans ࡞ࡻࡾ᳔∸࡫ࡡវ᯹ࡢࠉ୹࡞
఩Ὼ࡞ㄇᑙࡈࡿࡾ⬂Ꮔࡡฦ໩࠿ᘤࡀ㔘࡛࡝ࡾࠊࡐࡆ࡚ࠉῺᗐን໩࡞ࡻࡾ⬂Ꮔฦ໩ࡡฦᏄ࣒࢜ࢼࢫ࣑ࢅよ᪺ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙ࠉᠻࠍࡢ఩Ὼ࡞ࡻࡽⓆ⌟࠿ㄇᑙࡈࡿࡾ NIF (nuclear LIM-interacting interactor) 㐿ఎᏄࡡⓆ⌟โᚒᶭᵋ࡞ࡗ࠷࡙よᯊࡊࡒࠊGUS 㐿ఎᏄࢅ࣭ࣝ࣎ࢰ࣭࡞⏕࠷࡙ࠉḖ᥾࣒࣬࢞ࣚ࣬ᨭንࣈ࣓࣭ࣞࢰ࣭ࡡᶭ⬗ࢅよᯊࡊࠉ఩Ὼ࡞ᚺ➽ࡊࡒㄇᑙⓆ⌟࡞ࡢࠉ㌹෕㛜ጙⅤࡡ୕Ὦ 139-bp ࡞న⨠ࡌࡾ 7-nt 㒼า, GGACGAG,࠿ᚪ㡪࡚࠵ࡾࡆ࡛ࢅ᪺ࡼ࠾࡞ࡊࡒࠊࢤ࣭ࣜࢺ࣍ࢴࢠࢪ࡛ྞ௛ࡄࡼࡿࡒࢨࢪ࢙࣒ࣝࣤࢹࡢࠉ㎾⦍⛸ࡡ P. sojea ⏜ᮮࡡ
NIF 㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭୕࡞ࡵぜ࠷ࡓࡈࡿࡒࠊࢤ࣭ࣜࢺ࣍ࢴࢠࢪ≁␏Ⓩ࡞⤎ྙࡌࡾᰶࢰࣤࣂࢠ㈻࠿ EMSA Ἢ࡞ࡻࡽ᳠ฝࡈࡿࠉ⌟ᅹࢰࣤࣂࢠ㈻ࡡ⢥⿿ࢅモࡲ࡙࠷ࡾࠊ᭞࡞ࠉ㐗㉦Ꮔࡡᨲฝ࡛ࢤ࣭ࣜࢺ࣍ࢴࢠࢪࢅ௒ࡊࡒ NIF
㐿ఎᏄⓆ⌟ㄇᑙࡢࠉ⭯ࡡὮິᛮࢅ΅ᑛࡈࡎࡾ dimethyl sulfoxide ࡞ࡻࡽಀ㐅ࡈࡿࠉ⭯ࡡὮິᛮࢅቌຊࡈࡎࡾ
benzyl alcohol ࡞ࡻࡽᢒโࡈࡿࡒࠊ௧୕ࡡ⤎ᯕࡻࡽࠉ⬂Ꮔࡢࠉ⣵⬂⭯ࡡὮິᛮࡡ΅ᑛ࡞ࡻࡽῺᗐ఩ୖࢅវུࡊࠉ
ࡐࡿ࠿㐗㉦Ꮔࡡᨲฝ࡛ࢤ࣭ࣜࢺ࣍ࢴࢠࢪࢅ௒ࡊࡒ㐿ఎᏄⓆ⌟ࢅㄇᑙࡌࡾ࡛᥆ῼࡈࡿࡾࠊ
Activation of zoosporogenesis-specific genes in Phytophthora infestans involves a seven-nucleotide promoter
motif and cold-induced membrane rigidity.
Shuji Tani* and Howard S. Judelson (Dept. of Plant Pathology, UC Riverside. *present address: Grad. Sch. of Life &Env. Science, Osaka Pref. Univ.)

45

O-11
Monascus purpureus ࡞࠽ࡄࡾ࢜ࣄẐࢨࢹࣛࢼࣤࡡ⏍⏐ㄢ⟿ᅄᏄࡡᶭ⬗よᯊΰỀ೸㞕 1ࠉᮄୖᾀ 1ࠉỄ஬ྍ㑳 2ࠉொᖲ༜ஒ 1 㸝1 㜨ኬ࣬⏍∸ᅗ㝷ࢬࠉ2 ࣕ࣢࢝࢞㓬㓕ᢇ◂㸞⣊≟Ⳟ Monascus purpureus ࡢ⣒㯔࢜ࣄ࡛⛘ࡈࡿࠉཿࡂ࠾ࡼⓆ㓕㣏ဗ࠽ࡻࡦኮ↓╌Ⰵᩩ࡛ࡊ࡙㣏ဗᴏ⏲ฺ࡚⏕ࡈࡿ࡙࠷ࡾ㉝ⰅⰅ⣪ࡡ⏍⏐࡞⏕࠷ࡼࡿ࡙ࡀࡒࠊ㎾ᖳࠉⰅ⣪௧አ࡞ࡵ㧏⬙⾉⑍἖⒢࡛ⷾྜྷᵕࡡ⏍⌦Ὡᛮࢅ♟ࡌ࣓ࢻࢤࣛࣤࡷ GABA ࢅ஦ḗ௥ㅨ࡞ࡻࡽ⏍⏐ࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡽࠉ೸ᗛ㣏ဗ࡫ࡡᚺ⏕࠿᭿ᙽࡈࡿ࡙࠷ࡾࠊ
ࡊ࠾ࡊࠉᮇⳞࡢྜྷ᫤࡞ெ㛣࡞࡛ࡖ࡙᭯ᐐ࡝࢜ࣄẐ࡚࠵ࡾࢨࢹࣛࢼࣤࡵ⏍⏐ࡌࡾࡆ࡛࠾ࡼࠉ᭞࡝ࡾ᭯ຝฺ⏕࡞࠽࠷࡙ࠉࢨࢹࣛࢼࣤࡡ⏍ྙᠺᶭᵋࡡよ᪺ࠉโᚒ࠿ᚪ㡪࡛࡝ࡖ࡙࠷ࡾࠊᙔ◂✪ᐄ࡚ࡢࠉࢨࢹࣛࢼࣤ୹㦭᰹ࢅ⏍ྙᠺࡌࡾࠉ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ࢅࢤ࣭ࢺࡊ࡙࠷ࡾ㐿ఎᏄ pksCT ࡡࢠ࣭ࣞࢼࣤࢡ࡞ᠺຉࡊ࡙࠷ࡾࠊ⣊≟Ⳟ࡚ࡢࠉ஦ḗ௥ㅨ∸㈻ࡡ⏍ྙᠺ㛭㏻㐿ఎᏄ⩄ࡢࢠࣚࢪࢰ࣭ࢅᙟᠺࡊ࡙࠷ࡾࡆ࡛࠾ࡼࠉᮇⳞ࡞࠽࠷࡙ࡵ pksCT ࿔㎰࡞ࢨࢹࣛࢼࣤ⏍ྙᠺ㛭㏻㐿ఎᏄ⩄ࡢࢠࣚࢪࢰ࣭ࢅᙟᠺࡊ࡙࠷ࡾྊ⬗ᛮ࠿࠵ࡾ࡛⩻࠻ࡼࡿࡒࠊ࿔㎰㡷ᇡࡡࢨ࣭ࢠ࢙ࣤࢪよᯊࡡ⤎ᯕࠉpksCT ࡡ୕Ὦ⣑ 6 kb ࡡన⨠࡞ࠉDNA ⤎ྙ⬗ࢅ᭯ࡌࡾ 2Cys6 zinc finger motif ࢅᣚࡗ
regulator ࡛┞ྜྷᛮࢅ♟ࡌ ORF ࠿ぜฝࡈࡿࡒࠊࡆࡡ㐿ఎᏄ࡞ࡗ࠷࡙㐿ఎᏄ◒ቪ࡞ࡻࡾᶭ⬗よᯊࢅモࡲࡒ࡛ࡆࢀࠉ◒ቪᰬ࡞࠽࠷࡙ࢨࢹࣛࢼࣤ⏍⏐࠿⣑ 1/70 ࡞΅ᑛࡊࡒࡆ࡛࠾ࡼࠉࡆࡡ regulator ࡢࢨࢹࣛࢼࣤ⏍⏐ࡡὩᛮ໩࡞㛭୙ࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡒࠊᮇ◂✪ࡢࠉM. purpureus ࡞࠽࠷࡙ࡵ௙ࡡ⣊≟Ⳟ࡛ྜྷᵕ࡞ 2Cys6 zinc finger
motif ࢅᣚࡗ regulator ࠿Ꮛᅹࡊࠉ஦ḗ௥ㅨ∸㈻ࡡ⏍⏐࠿ㄢ⟿ࡈࡿ࡙࠷ࡾ᭩ิࡡሒ࿈ౚ࡚࠵ࡽࠉࡆࡡ regulator ࡡᶭ⬗โᚒࢅ㏳ࡋ࡙࢜ࣄẐ࡚࠵ࡾࢨࢹࣛࢼࣤࡡ⏍⏐㔖ࢅ΅ᑛࡈࡎࡾࡆ࡛࠿࡚ࡀࡾࡆ࡛࠿ึ᪺ࡊࡒࠊ
Functional analysis of regulator involved in citrinin production in Monascus purpureus.
Takeo Shimizua, Hiroshi Kinoshitaa, Shiro Nagaib, Takuya Nihiraa (aICBiotech, Osaka Univ., b Yaegaki Bio-industry,
Inc)

O-12
ⓉⰅ⭁ᮑⳞ⏜ᮮ P450/P450 ࣝࢱࢠࢰ࣭ࢭ⼝ྙࢰࣤࣂࢠ㈻ PcCYP17a ࡡᶭ⬗よᯊᚷỀඔ஼ 1ࠉᮿᓧⰟ⨶Ꮔ 1ࠉᗽᮆួႵ 2ࠉ᭯Ἁ❮ 2ࠉᜇᕖ༡ 2ࠉ๪▴༡஄ 1㸝1 ஐኬ㝌㎨ࠉ2 ࣒ࣜࢨࣔࣤ㸞ⓉⰅ⭁ᮑⳞ Phanerochaete chrysosporium ࡡࢣࢿ࣑ࣈࣞࢩ࢘ࢠࢹ࠿⤂஡ࡊࠉࡐࡡࢣࢿ࣑୯࡞ࡢ 154 ࡡ P450
㐿ఎᏄࢅ᭯ࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡈࡼ࡞ࠉP450 ࡛ P450 ࣝࢱࢠࢰ࣭ࢭࡡ⼝ྙࢰࣤࣂࢠ㈻࡞ࡗ࠷࡙ࡵ 7⛸Ꮛᅹࡊ࡙࠷ࡒࠊࡆࡿࡱ࡚ࠉ⼝ྙࢰࣤࣂࢠ㈻ࡡᶭ⬗࡞ࡗ࠷࡙ࡢࠉFusarium oxysporum ⏜ᮮࡡ P450foxy ࡷࠉᯜⲙⳞࡡ P450BM-3 ࡞ࡗ࠷࡙ሒ࿈ࡈࡿ࡙࠽ࡽࠉ㣤࿰⬙⫣㓗ࡡỀ㓗໩ (Ȱ-1ࠤȰ-3 న) ࢅ⾔࠹ࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊࡊ࠾ࡊ࡝࠿ࡼࠉP. chrysosporium ࡞࠽ࡄࡾᶭ⬗࡞ࡗ࠷࡙ࡢ᪺ࡼ࠾࡛࡝ࡖ࡙࠷࡝࠷ࠊࡐࡆ࡚ᮇ◂✪࡚ࡢࠉP. chrysosporium ⏜ᮮ P450/P450 ࣝࢱࢠࢰ࣭ࢭ⼝ྙࢰࣤࣂࢠ㈻࡚࠵ࡾ PcCYP17a ࡡᶭ⬗よᯊࢅ⾔ࡖࡒࠊኬ⭘Ⳟࢅᐙ୹࡛ࡊࡒ␏⛸Ⓠ⌟ࢅ⾔࠷ࠉᚋࡼࡿࡒࣛࢤࣤࣄࢻࣤࢹࢰࣤࣂࢠ㈻ࢅ⏕࠷࡙⅛⣪㙈㛏 9 ࠾ࡼ 18 ࡱ࡚ࡡ㣤࿰⬙⫣㓗ࢅᇱ㈻࡛ࡊ࡙ཬᚺࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ⅛⣪㙈㛏 9 ࠾ࡼ 15 ࡱ࡚ࡡ㣤࿰⬙⫣㓗࡞ࡗ࠷࡙ࡢ Ȱ-1ࠤȰ-6 నࢅỀ㓗໩ࡊࡒࡡ࡞ᑊࡊࠉ⅛⣪㙈㛏 16 ࠾ࡼ 18 ࡱ࡚ࡡ㣤࿰⬙⫣㓗࡞ࡗ࠷࡙ࡢ Ȱ-1ࠤȰ-4 నࢅỀ㓗໩ࡊࡒࠊࡱࡒࠉ⅛⣪㙈㛏 9 ࠾ࡼ 18 ࡱ࡚ࡡ㣤࿰⬙⫣࢓ࣜࢤ࣭ࣜ࡞ᑊࡊ࡙ࡵྜྷᵕ࡞Ề㓗໩ࡊࡒࠊࡈࡼ࡞ࠉࢻ
ࣆࢰࣝࣤࡷ࣊ࣤࢯ(a)ࣅࣝࣤ࡝࡜ⰶ㤮᪐໩ྙ∸࡞ࡗ࠷࡙ࡵỀ㓗໩ࡊࡒࠊࡆࡡࡆ࡛࠾ࡼࠉPcCYP17a ࡢࠉP450foxy
ࡷ P450BM-3 ࡛ᵋ㏸Ⓩ࡞␏࡝ࡖࡒᇱ㈻ヾㆉ㒂నࢅ᭯ࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ⌟ᅹࠉ㐿ఎᏄࡡⓆ⌟โᚒᶭᵋࢅྱࡴࠉࡈࡼ࡞レ⣵࡞よᯊࢅ⾔ࡖ࡙࠷ࡾࠊ

Molecular Characterization of a Fused Protein of P450 and P450 Reductase PcCYP17a from the White-rot
Fungus Phanerochaete chrysosporium
Motoyuki Shimizu1, Fumiko Matsuzaki1, Shinji Hirosue2, Akira Arisawa2, Hiroshi Tsunekawa2, Hiroyuki Wariishi1(1Faculty of Agriculture, Kyushu Univ., 2 Bioresource Laboratories, Mercian Co.)

46

O-13
ⓉⰅ⭁ᮑⳞ Phanerochaete chrysosporium ࡡ㧏✽⣪࠽ࡻࡦ఩✽⣪᮪௲ୖ࡞࠽ࡄࡾ⣵⬂አ
ࢰࣤࣂࢠ㈻ࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣛࣤࢡᑈᮇ ᐰࠉᚷỀඔ஼ࠉ๪▴༡஄ (ஐኬ㝌࣬㎨)
࠘┘Ⓩ࠙ⓉⰅ⭁ᮑᢰᏄⳞࡢᆀ⌣୕࡚᭩ࡵ㞬ฦよᛮࡡኮ↓㧏ฦᏄ࡚࠵ࡾࣛࢡࢼࣤࢅ။ୌ༟≺࡚↋ᶭ໩ྊ⬗࡝᚜⏍∸࡚࠵ࡾࠊⓉⰅ⭁ᮑⳞ Phanerochaete chrysosporium ࡢࣛࢡࢼࣤฦよ㓕⣪࡛ࡊ࡙ࠉ࣏࢛ࣤ࢝ࣤ࣋ࣜ࢞ࢨࢱ࣭ࢭ (MnP) ࠽ࡻࡦࣛࢡࢼ࢛ࣤ࣋ࣜ࢞ࢨࢱ࣭ࢭ (LiP) ࢅฦἢࡌࡾࠊࡆࡿࡼ㓕⣪ࡡⓆ⌟ࡢ✽⣪ᯜῤୖ࡞࠽࠷࡙ㄇ
ᑙࡈࡿࡾࡆ࡛࠿♟ࡈࡿ࡙࠷ࡾࠊࡆࡡࡆ࡛࠾ࡼࠉP. chrysosporium ࡡ✽⣪⃨ᗐ࡞౪Ꮛࡊࡒ⣵⬂አࢰࣤࣂࢠ㈻ࡡⓆ⌟โᚒᶭᵋ࡞⮾࿝࠿ᣚࡒࡿࡒࠊࡐࡆ࡚ࡱࡍࠉ㧏✽⣪ (HN) ࠽ࡻࡦ఩✽⣪ (LN) ᮪௲࡞࠽࠷࡙ࡐࡿࡑࡿฦἢࡈ
ࡿࡾࠉP. chrysosporium ⣵⬂አࢰࣤࣂࢠ㈻ࡡ⥑⨮Ⓩよᯊࢅ㛜ጙࡊࡒࠊ࠘᪁Ἢ࠽ࡻࡦ⤎ᯕ࠙HN ࠽ࡻࡦ LN ᮪௲ୖ࡚ㄢ⿿ࡊࡒ⣵⬂አࢰࣤࣂࢠ㈻ࢅ஦ḗඔ㞹Ẵὃິ࡞౩ࡊࡒ࡛ࡆࢀࠉ୦
᮪௲࡚Ⓠ⌟ࣂࢰ࣭ࣤ࠿ኬࡀࡂ␏࡝ࡖࡒࠊMALDI-TOF-MS よᯊ࡚ᚋࡒ࣋ࣈࢲࢺࣆ࢔࣭ࣤ࢝ࣈࣛࣤࢹࢅ⏕࠷࡙ࠉ
P. chrysosporium in silico protein database ࡞ᑊࡊ࡙ MASCOT ᳠⣬ࡌࡾࡆ࡛࡚ࢰࣤࣂࢠ㈻ࢅྜྷᏽࡊࡒࠊLN ୖ࡚ࡢࠉMnP1 ࡷ⣵⬂አ㐛㓗໩Ề⣪౩⤝⣌࡛⩻࠻ࡼࡿ࡙࠷ࡾ glyoxal oxidaseࠉaryl-alcohol oxidase ➴࠿ྜྷᏽࡈࡿࡒࠊ
ୌ᪁ HN ୖ࡞࠽࠷࡙ࡢࠉ々ᩐࡡ⢶㈻ຊỀฦよ㓕⣪ࠉglutaminaseࠉlipolytic enzymeࠉmannose 6-phosphataseࠉoxalate
decarboxylase ➴ࠉLN ୖ࡚ࡢぜࡼࡿ࡝࠾ࡖࡒ㓕⣪࠿ྜྷᏽࡈࡿࡒࠊ
Proteomic analysis of extracellular proteins from Phanerochaete chrysosporium grown on media with high andlow nitrogen concentrations
Hiroshi Teramoto, Motoyuki Shimizu, Hiroyuki Wariishi (Faculty of Agriculture, Kyushu Univ.)

O-14
Aspergillus oryzae ࢲࣞࢨࢻ࣭ࢭࡡ࢛ࣛࢥ࣏࣭໩࡛㓗Ὡᛮ໩ᶭᵋ
ኣ⏛⨮Ὂኯ㸡㞬ἴ ๙㸡ྚ⏛ Ꮟ 1㸡ୌᓞⱝ἖ 㸝๭౮ኬ㝌࣬⏍∸ᕝ㸡1 ᘧ๑ኬ࣬㎨Ꮥ⏍࿤㸞
A. oryzae ࢲࣞࢨࢻ࣭ࢭࡢ㓗ᛮ᮪௲࡚Ὡᛮ໩ࡌࡾ࡛࠷࠹≁␏Ⓩ࡝ᛮ㈻ࢅᣚࡗࠊᄿḗᵋ㏸ࡢ࣓࣌ᄿ㔖మ࡚Ὡᛮ୯ᚨࡢ 2 ࡗࡡ㖙ཋᏄ࡚࠵ࡾࠊࡆࡡ々㞟࡝ᵋ㏸ࢅᣚࡗࢲࣞࢨࢻ࣭ࢭࡡ㓗Ὡᛮ໩ᶭᵋࢅよ᪺ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡌ
ࡾࠊ୘Ὡᛮᆵࡡࣈࣞࢲࣞࢨࢻ࣭ࢭ (pro-TY)ࡢ pH 3.0 ࡞࠽࠷࡙Ὡᛮ໩ࡈࡿࡒ (acid-TY)ࠊࡱࡒ 0.01%ࡡ SDS ࡞ࡻࡽࢂࡍ࠾࡞Ὡᛮ໩ࡈࡿࡒࠊࢣࣜࢀ㐛࠾ࡼ pro-TY ࡛ acid-TY ࡡࢾ࢕ࢷ࢔᮪ࣥ௲࡞࠽ࡄࡾฦᏄ㈻㔖ࡢࡐࡿࡑࡿ
290kDa ࡛ 165kDa ࡚࠵ࡖࡒࠊCD ࢪ࣋ࢠࢹࣜࡡよᯊ࠾ࡼࠉ㓗Ὡᛮ໩࡞ࡻࡽ 2 ḗᵋ㏸࡞ን໩ࡢぜࡼࡿ࡝࠾ࡖࡒ
࠿ࠉ3 ḗᵋ㏸࡞ኬࡀ࡝ን໩࠿࠵ࡖࡒࠊࢹࣛࣈࢹࣆ࢒ࣤ⺧ක࡛ࢹࣛࣈࢹࣆ࢒ࣤ࡞⤎ྙࡌࡾ⺧කモⷾ
8-anilino-1-naphthalene sulfonic acid (ANS)࡞ࡻࡽࠉ㓗Ὡᛮ໩࡞ࡻࡽࢹࣛࣈࢹࣆ࢒ࣤṟᇱ࠿㟚ฝࡊ࡙࠷ࡾࡆ࡛࠿
♟ြࡈࡿࡒࠊࡆࡿࡼࡡ⤎ᯕ࠾ࡼ pro-TY ࡢ pH 3.0 ࡚ᄿ㔖మࡡฦᏄ࠿୘ྊ㏣Ⓩ࡞よ㞫ࡊ࡙஦㔖మ࡛࡝ࡽὩᛮ୯
ᚨࡡ㖙ཋᏄ࠿㟚ฝࡌࡾ࡛⩻࠻ࡼࡿࡾࠊ㑇ඔ≟ឺ࡞࠽ࡄࡾࢣࣜࢀ㐛࠾ࡼࠉὩᛮᆵ஦㔖మࡡࢦࣇࣗࢼࢴࢹ㛣ࡡ⤎ྙ࡞ࢩࢪࣜࣆ࢔ࢺ⤎ྙ࠿㛭୙ࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࢨࢪࢷ࢕ࣤṟᇱࢅ≁␏Ⓩ࡞⺧කᵾㆉࡌࡾ ABD-F ࡞
ࡻࡽࠉCys108 ࠿ฦᏄ㛣ࡡࢩࢪࣜࣆ࢔ࢺ⤎ྙ࡞㛭୙ࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊCys82 ࡢ㐗㞫ࢨࢪࢷ࢕࡚ࣤ࠵ࡽࠉ
ࡐࡡ௙ࡡ 6 ࡗࡡࢨࢪࢷ࢕ࣤṟᇱࡢฦᏄහ࡞࠽ࡄࡾࢩࢪࣜࣆ࢔ࢺ⤎ྙ࡞㛭୙ࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾࠊC108A ን␏㓕⣪ࡢ 68kDa ࡡ༟㔖మ (mono-C108A)࡛ 380kDa ࡡኣ㔖మ (oligo-C108A)ࡡ 2 ㏳ࡽࡡฦᏄ࠿⏍⏐ࡈࡿࡒࠊࡆ
ࡡ࠹ࡔ mono-C108A ࡞ࡢὩᛮ࠿ࡲࡼࡿࡍࠉ
oligo-C108A ࡢ㓗Ὡᛮ໩ࢅ⤊࡙㓕⣪Ὡᛮࢅࢂࡍ࠾࡞♟ࡊࡒࠊ
A. oryzaeࢲࣞࢨࢻ࣭ࢭࡡὩᛮ࡞ࡢ࢛ࣛࢥ࣏࣭໩࠿ᚪ㡪࡚࠵ࡾ࡛⩻࠻ࡼࡿࡾࠊ
Acid-activation and oligomerization of Aspergillus oryzae tyrosinase
Yota Tatara, Takeshi Namba, Takashi Yoshida1 Eiji Ichishima(Graduate School of Bioeng., Soka Univ. 1Faculty of Agric. and Life Sci., Hirosaki Univ.)

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⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾࣝࢪ࣎ࣤࢪ࣭ࣝ࢟ࣖࣝࢰ࣭ࡡよᯊⴏཋኬ♰ࠉ୷஬ῗୌ᭹ࠉຊ⸠㞖ኃࠉᑚᯐူኰࠉỀ㔕⊓㸝ྞኬ㝌࣬⏍࿤㎨Ꮥ㸞㸧ᠺฦโᚒ⣌ࡢ⣵Ⳟࡡࡲ࡝ࡼࡍ┷ᰶ᚜⏍∸࡞࠽࠷࡙ࡵᬉ㐚Ⓩ࡝⎌ሾᚺ➽᝗ሒఎ㐡ᶭᵋ࡚࠵ࡾࠊ࣓ࢸࣜ⣊≟Ⳟ A.nidulans ࡡ㸧ᠺฦโᚒ⣌ࢅ⥑⨮Ⓩ࡞よᯊࡌࡾࡆ࡛࡞ࡻࡖ࡙ࠉ⣊≟Ⳟ࡞࠽ࡄࡾᬉ㐚Ⓩ࡝⎌ሾᚺ➽࣒࢜ࢼࢫ࣑ࢅ᪺ࡼ࠾࡞ࡊࡒ࠷࡛⩻࠻ࡒࠊA.nidulans ࡡࢣࢿ࣑㒼าࢅㄢ࡬ࡾ࡛ࠉ㸧ᠺฦโᚒ⣌ࡡᵋᠺᅄᏄ࡚࠵ࡾࣃࢪࢲࢩ
ࣤ࢞ࢻ࣭ࢭ㸝HK㸞ࠉHPt ᅄᏄࠉࣝࢪ࣎ࣤࢪ࣭ࣝ࢟ࣖࣝࢰ࣭㸝RR㸞ࡢࡐࡿࡑࡿ 15 ⛸㢦ࠉ㸦⛸㢦ࠉ㸩⛸㢦Ꮛᅹࡌࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊᮇ◂✪࡚ࡢฝງ⣌࡚࠵ࡾࣝࢪ࣎ࣤࢪ࣭ࣝ࢟ࣖࣝࢰ࣭࡞↌Ⅴࢅࡊ࡯ࡽࠉ྘ RR ࡡ⏍⌦ᶭ⬗ࢅ᪺ࡼ࠾࡞ࡌࡾࡒࡴ࡞ 4 ⛸㢦ࡡ RR ◒ቪᰬࡡష⿿ࢅモࡲࡒࠊᚋࡼࡿࡒ◒ቪᰬࡢ࠷ࡍࡿࡵ⏍⫩㏷ᗐ࡞࠽࠷࡙㔕⏍ᰬ࡛ኬᕣࡢ࡝࠾ࡖࡒࠊࡊ࠾ࡊ SrrA ◒ቪᰬࡡᙟឺࡢ㔕⏍ᰬ࡛ⴥࡊࡂ␏࡝ࡽࠉ
ฦ⏍Ꮔᯮࡡᙟᠺ࡞␏ᖏ࠿ぜࡼࡿࠉ↋ᛮ⬂Ꮔ(conidia)ࡡ╌⏍ࡢⴥࡊࡂ఩ୖࡊ࡙࠷ࡒࠊࡈࡼ࡞ࠉ᭯ᛮ⏍Ṣჹᏻ࡚࠵ࡾࢠࣚ࢕ࢪࢹࢬࢲ࢓ࡡᙟᠺࡵふᐳࡈࡿ࡝࠾ࡖࡒࠊࡱࡒࠉSrrA ࡢ௧๑࡞ᠻࠍ࠿よᯊࡊࡒฦ⿛㓕ẍࡡ Prr1 ࡡ࣓࣌ࣞࢡ࡚࠵ࡽࠉPrr1 ࡛ྜྷᵕ࡞㓗໩ࢪࢹࣝࢪᚺ➽࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛࠿⩻࠻ࡼࡿࡒࠊ஢᝷㏳ࡽࠉ
SrrA ◒ቪᰬࡢ㓗໩ࢪࢹࣝࢪ࡞ᑊࡊ࡙វུᛮࢅ♟ࡊࠉᢘ㓗໩ᅄᏄ࡚࠵ࡾ࢜ࢰ࣭ࣚࢭࡡⓆ⌟࡞ࡵ㛭୙ࡊ࡙࠷ࡾࡆ࡛࠾ࡼࠉ㓗໩ࢪࢹࣝࢪᚺ➽ᶭᵋ࡞࠽࠷࡙㔔さ࡝ᙲ๪ࢅᯕࡒࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡆࡿࡼࡡࡆ࡛࠾ࡼ SrrA ࡢ⣊≟Ⳟࡡ⎌ሾᚺ➽࡞࠽࠷࡙ኣ㟻Ⓩ࡝࢙ࣆ࢘ࢠࢰ࣭࡚࠵ࡾ࡛⩻࠻ࡼࡿࠉࡐࡡ⏍⌦ᶭ⬗࡞ࡗ࠷࡙⩻ᐳࡌࡾࠊ
Aspergillus nidulans response regulator SrrA involved in oxidative stress response.
Daisuke Hagiwara, Junichirou Marui, Masashi Kato, Tetsuo Kobayashi, Takeshi Mizuno(Agriculture, Univ. of Nagoya)

O-16㯔Ⳟࣈࣞࢬࢴࢨࣤࢡࣈࣞࢷ࢓࣭ࢭ (KexB)ࡡḖ᥾࠿⣵⬂ባᵋ㏸࡞ཀྵ࡯ࡌᙫ㡢Ềㆺ἖ 1 㸡᳕ྞᮿᏄ 2, ఫ㔕ඔ᫓ 3, ᒜᙟὊᖲ 2, ῳ㑌๙ᚷ 4, 㜷㒂ᩏᝃ 2, ⏣⏛㞖஄ 5, ୯ᓞబ 2, ஫࿝ົஒ 1
㸝1 ᮶໪ኬ㝌㎨࣬⏍∸⏐ᴏ๭ᠺ, 2 ᮶໪ኬ㝌㎨࣬ᚺ⏍໩ࠉ3 㔘Ἁᕝኬࠉ4 ᩺₪ኬ࣬㎨ࠉ5 ⏐⥪◂㸞 ᠻࠍࡢࠉ㯔Ⳟࡡࣈࣞࢬࢴࢨࣤࢡ㓕⣪ KexB ࡡⳞమහ࡚ࡡᙲ๪ࢅよ᪺ࡌࡾࡒࡴ࡞ࠉkexB 㐿ఎᏄ◒ቪᰬࢅ㏸ᠺࡊࠉࡐࡡよᯊࢅ⾔ࡖ࡙ࡀࡒࠊࡐࡡ⤎ᯕࠉkexB ◒ቪᰬࡢࠉࣈ࣭ࣝࢹᇰ㣬࡚㢟ⴥ࡝ᙟឺᙟᠺ␏ᖏࢅ♟ࡊࠉࡐࡡᙟឺ␏ᖏࡡཋᅄࡡୌࡗ࡞⣵⬂ባࡡ⤣ྙᛮ࡞㛭୙ࡌࡾ cell integrity ࢨࢡࢻࣜఎ㐡⤊㊪ࡡᵋᠺⓏὩᛮ໩࠿♟ြࡈࡿࡒࠊࡐࡆ࡚ࠉࡆࡡ⤊㊪ࡡὩᛮ໩࠿ kexB ◒ቪᰬࡡ⣵⬂ባ࡞࡜ࡡᵕ࡝ᙫ㡢ࢅ୙࠻࡙࠷ࡾ࠾ࢅ᪺ࡼ࠾࡞ࡌࡾ┘Ⓩ࡚ࠉ㯔Ⳟ kexB ◒ቪᰬ⣵⬂ባࡡレ⣵࡝໩Ꮥᵋ㏸よᯊࢅモࡲࡒࠊ⣵⬂ባᵋ㏸よᯊࢅ⾔࠹࡞ࡒࡴ࡞ࠉkexB ◒ቪᰬཀྵࡦ㔕⏍ᰬࡡⳞమࢅ⇍Ề᢫ฝᚃࠉ࢓ࣜ࢜ࣛ᢫ฝ࡞ࡻࡽ࢓ࣜ࢜ࣛྊ⁈⏤ฦ࡛࢓ࣜ࢜ࣛ୘⁈⏤ฦ࡛࡞ฦ⏤ࡊࡒࠊࡐࡡ⤎ᯕࠉ kexB ◒ቪᰬࡡ⣵⬂ባࡢࠉ࢓ࣜ࢜ࣛ࡞ྊ⁈࡝⣵⬂ባ⏤ฦ࠿㔕⏍ᰬ࡛Ẓ㍉ࡊ࡙㟸ᖏ࡞ᑛ࡝࠷ࡆ࡛࠿
᪺ࡼ࠾࡞࡝ࡖࡒࠊ⌟ᅹࠉ྘⏤ฦ࡞ᑊࡌࡾᵋᠺ⢶ࡡᏽ㔖ཀྵࡦ࢞ࢲࢻ࣭ࢭࠉș-1,3-ࢡࣜ࢜ࢻ࣭ࢭ➴ࡡ㓕⣪ฌ⌦ࢅ⾔࠷ࠉᚋࡼࡿࡒ⏤ฦࢅよᯊࡊࠉኣ⢶ࡡฦᒪࡷ⤎ྙᵕᘟ࡞ࡱ࡚ን໩࠿࠵ࡾ࠾᳠ゞ୯࡚࠵ࡾࠊ
Perturbation of cell wall caused by defect of the KexB in Aspergillus oryzae.
Osamu Mizutani1, Matsuko Shiina1, Motoaki Sano2, Youhei Yamagata1, Takeshi Watanabe3, Keietsu Abe1, Masayuki
Machida4, Tasuku Nakajima1, Katsuya Gomi1(1 Tohoku Univ.,Grad.Sch.Agri.Sci, 2KIST, 3Niigata Univ., 4 AIST )

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⣊≟Ⳟ Aspergillus nidulans ࡡ࣐࢛ࢨࣤᵕࢺ࣒࢕ࣤࢅᣚࡗ஦ࡗࡡ࢞ࢲࣤྙᠺ㓕⣪
㸝CsmA, CsmB㸞ࡡ⣵⬂හᑻᅹ໩㒂నࠉᶭ ⬗Ⓩ┞㛭㛭౿ࡡよᯊ➁ୖ඼⏠ࠉᇷහ⿩஄ࠉኯ⏛᪺ᚠ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞 ࢞ࢲࣤࡢ⣊≟Ⳟࡡ⣵⬂ባࡡ୹さᵋᠺᠺฦࡡୌࡗ࡚࠵ࡽࠉࡐࡡ⏍ྙᠺࡢᙟឺᙟᠺࠉฦ໩࡞㔔さ࡝ᙲ๪ࢅᯕࡒ
ࡊ࡙࠷ࡾ࡛஢᝷ࡈࡿࡾࠊAspergillus nidulans ࡞ࡢࠉN ᮆ❻ഁ࡞࣐࢛ࢨ࡛ࣤ┞ྜྷᛮࢅ♟ࡌࢺ࣒࢕ࣤࢅᣚࡗࢰ࢕ࣈ
ࡡ࢞ࢲࣤྙᠺ㓕⣪ࢅࢤ࣭ࢺࡌࡾ csmAࠉ csmB ࡡ஦⛸㢦ࡡ㐿ఎᏄ࠿Ꮛᅹࡌࡾࠊࡆࡿࡱ࡚ࡡよᯊ࡞ࡻࡽࠉcsmA◒ቪᰬ࡛ csmB ◒ቪᰬࡡ⾪⌟ᆵࡡ㢦జᛮ࠿♟ࡈࡿࠉcsmA ࡛ csmB ࡡ஦㔔◒ቪ࠿ྙᠺ⮬Ṓ࡛࡝ࡾࡆ࡛࠿♟ࡈࡿ
࡙࠷ࡾ 1)ࠊࡱࡒ CsmA ࡢ࣐࢛ࢨࣤᵕࢺ࣒࢕ࣤࢅ௒ࡊ࡙࢓ࢠࢲࣤ⣵⬂㦭᰹౪ᏋⓏ࡞Ⳟ⣊඙❻ࡷ㝰ባᙟᠺ㒂న࡞ᑻᅹ໩ࡊࠉ⣵⬂ባྙᠺࢅ⾔࠹ࡆ࡛࠿♟ြࡈࡿ࡙࠷ࡾ 2)ࠊᮇ◂✪࡚ࡢࠉCsmB ࡡ⣵⬂හ࡚ࡡᑻᅹ໩㒂నࢅ᳠ゞࡌࡾࡒࡴࠉcsmB ࡡ ORF ࡡ C ᮆ❻ഁ࡞ 3 ࢤࣅ࣭ࡡ FLAG ࡡ tag ࠿ࡗ࡝࠿ࡖࡒᙟࡡࢰࣤࣂࢠ㈻ CsmB-FLAG ࢅ㔕⏍ᆵ CsmB ࡡ࠾ࢂࡽ࡞Ⓠ⌟࡚ࡀࡾᰬࢅష⿿ࡊࡒࠊ㛣᥃⺧කᢘమἪ࡞ࡻࡽよᯊࡊࡒ࡛ࡆࢀࠉCsmB-FLAG ࠿Ⳟ⣊඙❻ࠉ㝰ባᙟᠺ㒂నࡡ࢓ࢠࢲࣤ㎾എ࡞ᑻᅹࡌࡾᵕᏄ࠿ふᐳࡈࡿࡒࠊࡱࡒࠉCsmB ࡡ࣐࢛ࢨࣤᵕࢺ࣒࢕࡛ࣤ
࢓ࢠࢲࣤࣆ࢔࣒ࣚࣤࢹ࡛ࡡ⤎ྙࢅ in vitro ࡞࠽࠷࡙♟ࡊࡒࠊࡈࡼ࡞ࠉCsmA ࡛ CsmB ࡡᑻᅹ໩㒂నࡡẒ㍉ࡷ┞பష⏕࡞ࡗ࠷࡙ࡵよᯊࢅ⾔࠷ࠉ୦⩽ࡡᶭ⬗Ⓩ┞㛭㛭౿ࢅ᳠ゞࡊࡒࠊ1) ➁ୖࡼࠉ➠ 4 ᅂ⣊≟ⳞฦᏄ⏍∸Ꮥࢤࣤࣆ࢒ࣝࣤࢪさ᪠㞗 p. 7
2) Takeshita, N. et al. (2005) Mol. Biol. Cell 16, 1961-1970
Functional relevance between two chitin synthases CsmA and CsmB, each with a myosin motor-like domain, in
Aspergillus nidulans
Norio Takeshita, Hiroyuki Horiuchi, and Akinori Ohta
(Dept. of Biotechnol., Univ. of Tokyo)



O-18
Putative role of protein kinase C in the phosphorylation and multimerization of the Woroninbody protein, AoHex1, in Aspergillus oryzae
Praveen Rao JUVVADI, Jun-ichi MARUYAMA and Katsuhiko KITAMOTO(Dept. of Biotechnology, Univ. of Tokyo)
We previously characterized the Aspergillus oryzae Aohex1 gene encoding the major protein of the Woronin body anddemonstrated its plugging at the septal pore upon hypotonic shock by three-dimensional confocal microscopy1).
However, the regulatory mechanisms involved in the formation of the Woronin body remain unclear. In this report weshow the reduction in the dimeric and tetrameric forms of AoHex1 upon lambda-protein phosphatase treatment, whichindicated that AoHex1 phosphorylation in vivo may facilitate its multimerization. Concomitant with the presence of ahighly conserved predicted protein kinase C (PKC) phosphorylable site (Ser151), the recombinant AoHex1 was
phosphorylated in a PKC-dependent manner in vitro. Moreover, culturing A. oryzae in presence of chelerythrine, apotent PKC specific inhibitor, resulted in the reduction of the multimeric forms of AoHex1 implicating the relevance of
PKC-dependent phosphorylation of AoHex1 for its multimerization. Supporting these data, mutation of Ser151 to
Ala in AoHex1 caused reduced phosphorylation by PKC in vitro. While the Woronin bodies were visualized as dot-likestructures by expressing the dsred2-Aohex1, expression of the mutagenised construct, Aohex1 mt-S151A as dsred2 fusion
showed perturbed localization to ring-like structures, which were also evident under chelerythrine treatment of thedsred2-Aohex1 expressing strain.
1) Maruyama J. et al., (2005) Biochem. Biophys. Res. Commun. 33, 1081-8.
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㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ࢛࣭ࢹࣆ࢒ࢩ࣭ࡡฦ໩ࠉⓆ⏍࡫ࡡ㛭୙ⳝ㛣㝧ᚷ㸡ኬ᰷⏛Ꮼ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞࠘┘Ⓩ࠙ࣃࢹࡷ㓕ẍࢅࡢࡋࡴ࡛ࡌࡾ┷ᰶ⏍∸ࡢࠉᰜ㣬㣒㣱࡞ᚺ➽ࡊ࡙⣵⬂㈻ࡷ࢛ࣜ࢝ࢾࣚࢅᾦ⬂࡚ฦよࡊࠉ්ฺ⏕ࡌࡾ⣵⬂හฦよᶭᵋ࠿Ꮛᅹࡌࡾࠊࡱࡒ㎾ᖳࠉ࢛࣭ࢹࣆ࢒ࢩ࣭࠿ᰜ㣬㣒㣱᫤ࡡ⏍Ꮛᡋ␆࡛ࡊ࡙ࡓࡄ࡚࡝
ࡂࠉ㏳ᖏ᫤ࡡ⣵⬂හ௥ㅨᅂ㌹ࠉฦ໩ࡷⓆ⏍ࠉඞ␷ᶭᵋࠉវ᯹⑋ཋమ࠾ࡼࡡ㜭ᚒࠉ⣵⬂Ṓ࡝࡜࡞ࡵ㛭୙ࡌࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊᮇ◂✪࡚ࡢࠉA. oryzae ࢅࡢࡋࡴ࡛ࡌࡾ⣊≟Ⳟ࡞࠽ࡄࡾ࢛࣭ࢹࣆ࢒ࢩ࣭࡞㛭ࡌࡾ▩ぜࢅᚋࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙ࠉ࢛࣭ࢹࣆ࢒ࢩ࣭࡞࠽࠷࡙㔔さ࡝ᶭ⬗ࢅᯕࡒࡌฝⰾ㓕ẍ ATG8 ࣓࣌ࣞࢡ㐿ఎᏄ Aoatg8ࡡ◒ቪ࡞ࡻࡾ࢛࣭ࢹࣆ࢒ࢩ࣭ࡡ⏍⌦ᶭ⬗ࡡよᯊࢅ⾔ࡖࡒࠊ࠘᪁Ἢཀྵࡦ⤎ᯕ࠙ࡆࡿࡱ࡚࡞ᠻࠍࡢࠉEGFP-AoAtg8 ࠽ࡻࡦ DsRed2-AoAtg8 ⼝ྙࢰࣤࣂࢠ㈻ࡡᑻᅹよᯊ࡞ࡻࡽA. oryzae ࡞࠽ࡄࡾ࢛࣭ࢹࣆ࢒ࢩ࣭ࡡᏋᅹࢅ♟ࡊࡒ㸦㸞ࠊ௑ᅂࠉAoatg8 ◒ቪᰬࢅషᠺࡊࠉࡐࡡ⾪⌟ᆵࡡよᯊࢅ⾔ࡖࡒࠊAoatg8 ◒ቪᰬࡢẴ୯Ⳟ⣊࠽ࡻࡦฦ⏍Ꮔࡡᙟᠺ࡞Ḗ᥾࠿ぜࡼࡿࡒࠊࡱࡒ᭩ᑚᇰᆀ࡞࠽࠷࡙ࡢࢂࡍ࠾࡝࠿
ࡼ⏍⫩㜴ᐐ࠿ふᐳࡈࡿࡒࠊࡆࡡ◒ቪᰬࡡ⣵⬂㈻࡞ DsRed2 ࢅⓆ⌟ࡈࡎࠉᰜ㣬㣒㣱࡞࠽ࡄࡾᾦ⬂࡫ࡡཱིࡽ㎲ࡲ
ࢅふᐳࡊࡒ࡛ࡆࢀࠉAoatg8 ◒ቪᰬ࡚ࡢ DsRed2 ࡡཱིࡽ㎲ࡲ࠿ふᐳࡈࡿ࡝࠾ࡖࡒࠊࡆࡿࡢ Aoatg8 ◒ቪᰬ࠿ᐁ㝷࡞࢛࣭ࢹࣆ࢒ࢩ࣭ࢅḖ᥾ࡊ࡙࠷ࡾࡆ࡛ࢅ♟ࡊ࡙࠽ࡽࠉAoatg8 ࠿࢛࣭ࢹࣆ࢒ࢩ࣭࡞ᚪ㡪࡚࠵ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡱࡒࠉࢲ࢓࣐ࣤࣈ࣓࣭ࣞࢰ࣭ୖ࡚ AoAtg8 ࢅ Aoatg8 ◒ቪᰬ࡞࠽࠷࡙Ⓠ⌟ࡈࡎࡒ┞⿭ᰬࢅふᐳࡊࡒ⤎ᯕࠉࢲ࢓࣐⃨ࣤᗐ౪ᏋⓏ࡞Ẵ୯Ⳟ⣊࠽ࡻࡦฦ⏍Ꮔࡡᙟᠺ࠿ᅂᚗࡌࡾࡆ࡛࠿ふᐳࡈࡿࡒࠊ௧୕ࡡ⤎ᯕࡻࡽࠉA. oryzae࡞࠽࠷࡙࢛࣭ࢹࣆ࢒ࢩ࣭࠿ฦ໩ࡷⓆ⏍࡞㛭୙ࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ⌟ᅹࠉࡆࡡ┞⿭ᰬ࠾ࡼฦ⏍Ꮔࢅᅂ཭ࡊࠉ
࢛࣭ࢹࣆ࢒ࢩ࣭࠿Ⓠⰾ࡞୙࠻ࡾᙫ㡢࡞ࡗ࠷࡙ࡵ᳠ゞ୯࡚࠵ࡾࠊ
㸦㸞ⳝ㛣ࡼࠉ➠ 4 ᅂ⣊≟ⳞฦᏄ⏍∸ࢤࣤࣆ࢒ࣝࣤࢪさ᪠㞗 p.67
Evidences for autophagy during differentiation and development of A. oryzae
Takashi Kikuma, Mamoru Ohneda, Manabu Arioka, Katsuhiko Kitamoto(Dept. of Biotechnology, Univ. of Tokyo)

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࣑࢟㢦㉝࢜ࣄ⑋Ⳟ Fusarium graminearum ࡡⓆⰾ⬂Ꮔ࡞࠽ࡄࡾࣛࣤ㓗໩ࢰࣤࣂࢠ ㈻ࡡよᯊ⏍㥎༜ஒ㸡㡪㈙ᬍ஁*㸡㕝ᮄᚥ*㸡H. C. Kistler. **㸡Ⓤ⏣‫‮‬᭹㸝ᒪኬ࣬ᚺ⏍㸡*ᒪኬ࣬⏍࿤ࢬ㸡**Univ. of Minnesota㸞࣑࢟㢦㉝࠾ࡦ⑋Ⳟ Fusarium graminearmu ࡢ✈∸࡞⑋ᐐ࡛࠾ࡦẐỗ᯹ࢅ㉫ࡆࡌࡆ࡛࡚ኬࡀ࡝ၡ㢗࡛࡝ࡖ࡙࠷ࡾ
⣊≟Ⳟ࡚࠵ࡾࠊ2003 ᖳࠉᮇⳞࡢࡐࡡ㔔さᛮ࠾ࡼ᳔∸⑋ཋ⣊≟Ⳟ࡚ 2 ␊┘࡞ 36Mbp ࠾ࡼ࡝ࡾࢣࢿ࣑ࡡධሲᇱ㒼า࠿Ửᏽࡈࡿࠉ⣑ 11,640 ಴ࡡ㐿ఎᏄ࠿ぜฝࡈࡿࡒࠊⁿ⩽ࡼࡢវ᯹ิ᭿࡞ F. graminearum ࡚ᶭ⬗ࡊ࡙࠷ࡾ㐿ఎᏄࢅ᪺ࡼ࠾࡞ࡌࡾࡒࡴⓆⰾฦ⏍⬂Ꮔ࡞࠽ࡄࡾࣈࣞࢷ࢛࣭࣑よᯊࢅ⾔ࡖࡒ⤎ᯕࠉࢰࣤࣂࢠ㈻࡚ࣝ࣊ࣜᑛ࡝ࡂ࡛
ࡵ 188 ಴ࡡ㐿ఎᏄⓆ⌟ࢅヾࡴࡒ㸝ᖲᠺ 17 ᖳᗐ ᪝ᮇ᳔∸⑋⌦Ꮥఌኬఌ㸞ࠊࡐࡆ࡚ᮇ◂✪࡚ࡢ⩳ゼᚃಞ㣥ࡡୌ⛸࡚࠵ࡾࢰࣤࣂࢠ㈻ࡡࣛࣤ㓗໩࡞ࡗ࠷࡙ㄢ࡬ࡒࠊࡱࡍⓆⰾฦ⏍⬂Ꮔ࠾ࡼࢰࣤࣂࢠ㈻ࢅ᢫ฝࡊࠉ஦ḗඔࢣࣜ㞹Ẵὃິ࡚ฦ㞫ࡊࡒࠊḗ࡞ࢲࣞࢨࣤࠉࢬࣛࣤࠉࢪ࢛ࣝࢼࣤṟᇱ࡞⤎ྙࡊࡒࣛࣤ㓗ᇱࡡ᳠ฝ࠿ྊ⬗࡝モⷾࢅ⏕࠷࡙ࢣࣜࢅ᯹Ⰵࡊࡒࠊࡐࡡ⤎ᯕࠉ⣑ 410 ಴ࡡࢰࣤࣂࢠ㈻ࢪ࣎ࢴࢹ୯ࠉ37 ಴ࡡࢪ࣎ࢴࢹ࠿ࣛࣤ㓗໩ࢰࣤࣂࢠ㈻࡛
ࡊ࡙᳠ฝࡈࡿࡒࠊࡆࡿࡱ࡚࡞షᠺࡊࡒ 2DE-reference map㸝಴ࠍࡡࢰࣤࣂࢠ㈻ࢪ࣎ࢴࢹ࡞㐿ఎ᝗ሒ࠿௛ຊࡈࡿ
ࡒ஦ḗඔࢣࣜ㞹Ẵὃິാ㸞࡞↯ྙࡊ࡙ࡐࡿࡼࡢ௙ࡡ᳔∸⑋ཋᛮ⣊≟Ⳟ࡚⑋ཋᛮ࡞㛭౿࠿࠵ࡾ࡛ࡈࡿ࡙࠷ࡾ
CAP20ࠉ྘⛸ dehydrogenaseࠉworonin body protein precursorࠉelongation factor ࡝࡜ࢅྱࡳࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ
ࡒࠊ
Phosphoprotein analysis of germed conidiospores of Fusarium graminearum
Takuya Ikoma, *Haruhisa Suga, *Toru Suzuki, **H. C. Kisler, Mituro Hyakumachi(Faculty of Applied Biological Sciences, Gifu Univ *LSRC, Gifu Univ. **Univ. of Minnesota)

P-2ࢲࣛ⏐ࢹ࣏ࢹᒌ㔕⏍⛸࠾ࡼฦ㞫ࡊࡒ㟸⑋ཋᛮ Fusarium oxysporum ࡡฦᏄ⣌⤣よᯊᒱ㒂᪺Ꮔ, ▴ᕖᬰᏄ*, ᕖ㒂┶Ⓡ**, ඡ⋚ᇱୌ㑳***, ᑈᒱ ᚥ, ᭯Ờ ງ 㸝᮶ா㎨ᕝኬ࣬*⌟㣏ဗฦᯊࢬࣤࢰ
࣭࣬**࣠ࢨࣤࢹࣤᕗኬ䝿***㫵ཱིኬ㸞ࢹ࣏ࢹཋ⏐ᆀ㎾എ࡛᥆ᏽࡈࡿ࡙࠷ࡾࢲࣛභ࿰ᅗ໪㒂ᆀᇡ࠾ࡼࠉ 2002 ᖳ 12 ᭮࠽ࡻࡦ 2004 ᖳ 3 ᭮࡞ࢹ࣏ࢹᒌ㔕⏍⛸ ( Lycopersicon pervanum, L. chilens ) ࠽ࡻࡦ᰺ᇰ⛸ ( L. esculentum ) ࢅ᤿㞗ࡊࡒࠊࡆࡿࡼ᳔∸మࡡᯕᐁහ㒂ࠉᯕᐁአ㒂ࠉⱴአ㒂ࠉⱴහ㒂ࠉⴝࠉ᰷࠾ࡼゝ 562 ᰬࡡ⣊≟Ⳟࢅฦ㞫ࡊࠉྜྷᏽࢅ⾔ࡖࡒ࡛ࡆࢀࠉAlternalia
ᒌⳞࠉ Aspergillus ᒌⳞࠉ Cladosporium ᒌⳞࠉ Fusarium ᒌⳞࠉ Geotrichum ᒌⳞࠉ Mucor ᒌⳞࠉRhizopus
ᒌⳞࠉ Papulaspora ᒌⳞࠉPenicillium ᒌⳞࠉ Phoma ᒌⳞࠉ Trichoderma ᒌⳞࠉ Ulocladium ᒌⳞ࠿ぜฝࡈࡿࡒࠊ Fusarium ᒌⳞ࡞ࡗ࠷࡙ࡢ⛸ࢅྜྷᏽࡊࠉ F. oxysporum 39 ᰬࡡࢹ࣏ࢹⴆ෺⑋Ⳟ࣭ࣝࢪ᳠ᏽ⏕ࢹ࣏ࢹဗ⛸࡞ᑊࡌࡾ⑋ཋᛮࢅㄢᰕࡊࡒ࡛ࡆࢀࠉ࠷ࡍࡿࡵ⑋ཋᛮࢅ♟ࡈࡍࠉ㟸⑋ཋᛮ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡒࠊࡆࡿࡼࡡⳞᰬ
ࡡ rDNA IGS 㡷ᇡࡡሲᇱ㒼าࢅỬᏽࡊࠉࡌ࡚࡞Ⓡ㘋ῥࡲࡡ F. oxysporum ྘ฦ໩ᆵᰬࡡሲᇱ㒼า᝗ሒ ( ᕖ㒂ࡼ, 2005 ) ࡛Ẓ㍉ࡊࠉฦᏄ⣌⤣よᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉࢹ࣏ࢹᒌ㔕⏍⛸࠽ࡻࡦ᰺ᇰ⛸⏜ᮮࡡ F. oxysporumⳞᰬࡢᴣࡠ F. oxysporum f. sp. lycopersici ( FOL ) ᰬ࡞ࡻࡖ࡙ᵋᠺࡈࡿࡾࢠࣚࢪࢰ࣭ࡡ㎾എ࡞న⨠ࡊࠉ௙ฦ໩ᆵ
Ⳟᰬࡻࡽࡵ FOL ࡞㎾⦍࡚࠵ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡱࡒ 2005 ᖳ 7 ᭮࡞ࠉ࣒࢞ࢨࢤ࡚ࠉ㔕⏍⛸࠾ࡼ᰺ᇰ⛸࡫ࡡ⛛⾔᭿࡞࠵ࡖࡒ࡛᝷ᏽࡈࡿ࡙࠷ࡾࢹ࣏ࢹ ( L. esculentum var. cerasiforme ) ࢅ᤿㞗ࡊࠉྜྷᵕ࡞ F. oxysporumࢅฦ㞫ࡊࠉ⌟ᅹࡆࡿࡼࡡⳞᰬ࡞ࡗ࠷࡙ࡵ࠵ࢂࡎ࡙⣌⤣よᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
Genealogical analyses of non - pathogenic Fusarium oxysporum isolated from Wild Lycopersicon spp. in Chile.

Akiko Okabe, *Nobuko Ishikawa, **Masato Kawabe, ***Motoichiro Kodama, Tohru Teraoka, Tsutomu Arie
(Tokyo Univ. of Agric. & Tech., *Japan Food Res. Lab., **Washington state Univ., ***Tottori Univ.)

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Gibberella fujikuroi Ꮔᄙ⬂Ꮔ࡞࠽ࡄࡾᙟ㈻ࡡฦ㞫࡛ᇰ㣬୕ΰ࡞ࡻࡾᏄᄙṾᙟᠺㄇᑙᐣᶌຊዄ㸡ᑈᒱ ᚥ㸡᭯Ờ ງ 㸝㎨ᕝኬ࣬㎨㸞
 ࢦࢹࢗ࢞ࣄࡊࡺ࠹㢄⭁ᩃ⑋Ⳟ Gibberella fujikuroi mating population B(anamorph : Fusarium sacchari)ࡢஹ㒼ᆵ
㸝mating type㸞ࡡ␏࡝ࡾ(MAT1-1㸡MAT1-2)Ⳟᰬ㛣࡚ஹ㒼ࢅ⾔࠷ࠉᏰධୠ௥ࢅᙟᠺࡌࡾ heterothallic ࡝ᏄᄙⳞ࡚࠵ࡾࠊG. fujikuroi FGSC 7610 ᰬ(MAT1-2㸡㞜ᛮ୘⛩)࠽ࡻࡦ FGSC 7611 ᰬ(MAT1-1㸡୦ᛮ⛩ᛮ)࡞ EGFP Ⓠ⌟
࢜ࢬࢴࢹ࡛ࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤ B ⪇ᛮ࢜ࢬࢴࢹࢅྜྷ᫤࡞ᑙථࡊ࡙ᚋࡼࡿࡒᙟ㈻㌹ᥦమ࡛㔕⏍ᰬ㛣࡛࡚ஹ㒼ࡊ࡙ᙟᠺࡈࡿࡒᏄᄙ⬂Ꮔࢅ༟⬂Ꮔฦ㞫ᚃࠉ3 ᙟ㈻㸝GFP/ࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤ⪇ᛮ/ஹ㒼ᆵ㸞࡞ࡗ࠷࡙㐿ఎⓏよᯊࢅ⾔ࡖࡒ࡛ࡆࢀࠉ㔕⏍ᆵ FGSC 7610 ᰬࢅ㞕ഁࠉᙟ㈻㌹ᥦమ FGSC 7611 ᰬࢅ㞜ഁ࡛ࡊ࡙ஹ㒼ࡌࡾ࡛ࠉ࡮࡯ 1㸯1 ࡞
྘ᙟ㈻࠿ฦ㞫ࡊࡒ࠿ࠉᙟ㈻㌹ᥦమ FGSC 7610 ᰬࢅ㞕ഁࠉ㔕⏍ᆵ FGSC 7611 ᰬࢅ㞜ഁ࡞ࡌࡾ࡛ࠉ྘ᙟ㈻࠿ᚪࡍ
ࡊࡵ⌦ㄵ㏳ࡽ࡞ฦ㞫ࡊ࡝࠾ࡖࡒ㸝2004 ᖳᗐሒ࿈㸞ࠊ්ᗐࠉࡐࡡฦ㞫Ẓࢅ⢥ᰕࡌࡾ࡛࡛ࡵ࡞ࠉ⫴ᬊ࡛࡝ࡾஹ㒼ᣪິ࡞ࡗ࠷࡙⩻ᐳࡊࡒࠊࡱࡒࠉ㏳ᖏࡡஹ㒼᫤࡞㞕࡛ࡊ࡙⏕࠷ࡾ㞕ᰬ⬂Ꮔᠩ⃦ᾦ࡞௥࠻࡙ࠉ㞕ᰬࡡᇰ㣬୕ΰ㸝℈㐛ᇰ㣬ᾦ㸞ࢅฌ⌦ࡊࡒሔྙ࡞ࡵᏄᄙṾᙟᠺ࠿ㄇᑙࡈࡿࡒࠊࡈࡼ࡞ࠉᮇⳞࡡ㎾⦍⛸࡚࠵ࡾ Fusarium oxysporum㸝୘Ᏸධᛮ㸞ࡡ⬂Ꮔᠩ⃦ᾦࢅฌ⌦ࡊࡒሔྙ࡞ࡵࠉஹ㒼ᆵࡡ␏࡝ࡾⳞᰬྜྷኃࡡ⤄ྙࡎ㛣࡞࠽࠷࡙ࡡࡲᏄᄙṾᙟᠺ࠿ㄇᑙࡈࡿࡒࠊ
Segregation of phenotypes in ascospores of Gibberella fujikuroi and induction of barren perithecia formation by
culture supernatantKana Togashi㸡 Tohru Teraoka㸡Tsutomu Arie
(Tokyo Univ. of Agric.㸛 Tech.)P-4ࣁ࣭ࢷ࢔ࢨ࣑ࣛࢗⴆ෺⑋Ⳟ Verticillium longisporum ࡡ PCR ᳠ฝ࡛ฦᏄ⣌⤣よᯊ᩟⸠⚵ᠺ㸦㸡᲻ἃ⧂㸦㸡㒿஬Ᏻ㸧㸡⁵ཋᑋᙢ㸧㸡⸠ᮟ┷㸦 㸝㸦᮶Ὂኬ࣬⏍࿤㸡㸧⩄㤷㎨ᢇࢬ࣬⏍⏐⎌ሾ㸞ᅰቫ᳔∸⑋ཋⳞ Verticillium ࡞ࡻࡾష∸⿍ᐐࡢࠉ୹࡞ V. dahliae ࡛ V. albo-atrum ࡞ࡻࡾ࡛ࡈࡿ࡙࠷ࡾ࠿ࠉ⩄㤷┬ࡡᎶᜂᆀ༇࡚ࡢࠉ࢓ࣇࣚࢻ⛁ష∸࡞ᙁ࠷⑋ཋᛮࢅ♟ࡌ V. longisporum ࠿ V. dahliae ࡛࡛ࡵ࡞ၡ㢗࡛࡝ࡖ࡙࠷ࡾࠊྜྷᆀ༇࠾ࡼฦ㞫ࡊࡒ V. longisporum ࡡ CA9 ᰬࢅ⏕࠷࡙ rDNA ࡡ㒼าࢅỬᏽࡊࠉV. dahliae ࡷ V. albo-atrum࡛Ẓ㍉ࡊࡒ࡛ࡆࢀࠉITS 㡷ᇡࡢࡳࡊࢀ V. albo-atrum ࡛┞ྜྷᛮ࠿㧏࠷ࡆ࡛ࠉ18S 㡷ᇡ࡞ 841bp ࡡ≁␏Ⓩ࡝᣼ථ㒼าࢅࡵࡗࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡆࡿࡼࡡ᝗ሒࢅࡵ࡛࡞ V. longisporum ࢅ PCR ᳠ฝࡌࡾ 2 ࢬࢴࢹࡡࣈࣚ࢕࣏
࣭ࢅషᠺࡊࠉᅰቫ୯ࡡ CA9 ᆵ V. longisporum ࢅ᳠ฝࡊࠉᏽ㔖ࡵྊ⬗࡝ࣛ࢓ࣜࢰ࢕࣑ PCR ⣌ࢅᵋ⠇ࡊࡒࠊࡊ࠾
ࡊࠉྜྷᆀ༇࠾ࡼ V. longisporum ࡛ࡊ࡙ฦ㞫ࡊࡒ 8 ᰬࢅ PCR チ᩷ࡊࡒ࡛ࡆࢀࠉ⣑༖ᩐ࠿ V. dahliae ࡛チ᩷ࡈࡿࡒࠊ
ࡆࡿࡼࡡ Vd ᆵᰬࡡᙟឺᏕⓏ≁ᚡ㸝⬂Ꮔ㛏ᙼࡷⳞᰶᙟ≟㸞࠽ࡻࡦ⏍໩ᏕⓏ≁ᚡ㸝࣎ࣛࣆ࢘ࢿ࣭࢛ࣜ࢞ࢨࢱ࣭
ࢭὩᛮ㸞ࡢࠉ࠷ࢂࡹࡾ V. dahliae ࡛ࡢ␏࡝ࡽ V. longisporum ࡡࡐࡿ࡛ୌ⮬ࡊࡒࠊCA9 ᆵ࠽ࡻࡦ Vd ᆵ࡛࠷࠹ 2⛸㢦ࡡⳞᰬࡡ㐿ఎⓏ⫴ᬊࢅẒ㍉ࡌࡾࡒࡴ࡞ࠉRAPD Ἢよᯊࡷ࣐ࢹࢤࣤࢺࣛ࢓㐿ఎᏄࡡ PCR-RFLP Ἢ࡞ࡻࡾよᯊࢅ⾔࡝ࡖࡒࠊࡆࡿࡼࡡ⤎ᯕ࠾ࡼࠉVd ᆵᰬࡢࠉCA9 ᆵᰬ࡛භ㏳ࡡኣᆵࢅ♟ࡌሔྙ࡛ V. dahliae ࡛භ㏳ࡡኣᆵࢅ♟ࡌሔྙ࠿࠵ࡾࡆ࡛࠿ึ᪺ࡊࡒࠊࡆࡿࡼࡡࡆ࡛࠾ࡼࠉᎶᜂᆀ༇ࡡ V. longisporum ࡞ࡢࠉ㐿ఎⓏ࡝⫴ᬊ࠿␏࡝ࡾ࡛ᛦࢂࡿࡾ 2 ⛸㢦ࡡᰬ࠿ᏋᅹࡊࠉVd ᆵᰬࡢ V. longisporum ࡛ V. dahliae ࡛୦᪁ࡡ㐿ఎⓏ⫴ᬊࢅࡵࡗྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊ
Molecular characterization of two types of Verticillium longisporum isolates from cabbage fields in Tsumagoi
Hidenari Saito1, Saori Mori1, Hiroshi Sakai2, Toshihiko Urushibara2, Makoto Fujimura1㸝1Fac.of Life Sci.,Toyo Univ.;2

Gunma Pref. Agri. Tech. Center㸞

52

P-5
ࢗࣛ㢦⅛⑃⑋Ⳟࡡ ClaCWH41 㐿ఎᏄࡢᐙ୹౴ථ࡞ᚪ㡪࡚࠵ࡽ౴ථჹᏻࡡ⣵⬂ባྙᠺ࡞㛭୙ࡌࡾᮿ஬⌦ᜠ࣬ᐋᆀಆᙢ࣬㎱ ඔெ࣬㎱ᒜ ᙪୌ࣬Richard O'Connell࣬Ⓣ▴ཪ⣎㸟࣬஁ಕᗛ஄ 㸝ாᗋኬ㝌㎨࣬ᒱᒜ
ኬ㎨㸟㸞
ࢗࣛ㢦⅛⑃⑋Ⳟࡡ௛╌ჹᙟឺฦ໩ࡢᐙ୹౴ථ࡞ᚪ㡪࡚࠵ࡾ㸣௑ᅂ㸡REMI Ἢ࡞ࡻࡽషฝࡊࡒ௛╌ჹᙟឺᙟᠺን␏ᰬ P51 ࡡ◒ቪ㐿ఎᏄࡡྜྷᏽ࡛ᛮ≟よᯊࢅ⾔࡝ࡖࡒ㸣ᮇን␏ᰬࡡን␏㡷ᇡࡡよᯊࢅ⾔ࡖࡒ࡛ࡆࢀ㸡ฝⰾ㓕ẍ࡞࠽࠷࡙⣵⬂ባྙᠺ࡞㛭୙ࡌࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾ CWH41 㐿ఎᏄ࡛㧏࠷┞ྜྷᛮࢅ♟ࡊ㸡ᮇ㐿ఎᏄࢅ
ClaCWH41 ࡛࿤ྞࡊࡒ㸣ḗ࡞ AtMT Ἢࢅ⏕࠷࡙ clacwh41 ◒ቪᰬࢅషฝࡊ㸡ᛮ≟よᯊࢅ⾔ࡖࡒ㸣clacwh41 ◒ቪᰬࡢᘽ⑋ཋᛮࢅ♟ࡊ㸡ெᕝࢬ࣭ࣜࣞࢪ⭯࡞ᑊࡌࡾ∸⌦Ⓩ౴ථ⬗ࡡ఩ୖࢅ♟ࡊࡒ㸣ࢪࣚ࢕ࢺ࢝ࣚࢪ୕࡞࠽ࡄࡾ
௛╌ჹᙟឺࢅふᐳࡊࡒ࡛ࡆࢀ㸡㔕⏍ᰬ࠿⌣ᙟࡡ௛╌ჹࢅᙟᠺࡌࡾࡡ࡞ᑊࡊ㸡◒ቪᰬࡢ␏ᖏ࡝௛╌ჹࢅᙟᠺࡊ
࡙࠷ࡒ㸣㞹Ꮔ㢟᚜㙶ࢅ⏕࠷࡙Ⓠⰾ⬂ᏄཀྵࡦⳞ⣊ࢅ㔕⏍ᰬ࡛Ẓ㍉ࡊࡒ࡛ࡆࢀ㸡Ⳟ⣊ᙟឺࡢ㔕⏍ᰬ࡛ྜྷᵕ࡚࠵ࡖ
ࡒ㸣ࡊ࠾ࡊ࡝࠿ࡼ㸡௛╌ჹᙟᠺ᫤࡞࠽ࡄࡾ◒ቪᰬࡡ௛╌ჹཀྵࡦⓆⰾ⬂Ꮔࡢ⣵⬂ባ࠿⫟ኬࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ
࠾࡛࡝ࡖࡒ㸣௧୕ࡡ⤎ᯕ࠾ࡼ㸡౴ථჹᏻࡡ⣵⬂ባྙᠺࡡ␏ᖏ࠿㸡௛╌ჹࡡ౴ථᶭ⬗ࡡḖ᥾࡞㛭㏻ࡌࡾྊ⬗ᛮ࠿⩻࠻ࡼࡿࡒ㸣The ClaCWH41 Gene is Essential for Penetration of Host Tissue and Cell Wall Formation during Germling
Development in Colletotrichum lagenarium.
Rie Matsui, Toshihiko Miyaji,Gento Tsuji, Shouichi Tsujiyama, Richard O'Connell, Tomonori Shiraishi and Yasuyuki
Kubo 㻃 (Univ. of Kyoto Pref, Univ. of Okayama.)

P-6
GATEWAY ࢨࢪࢷ࣑ࢅฺ⏕ࡊࡒ⣊≟Ⳟ⏕ 㐿ఎᏄ◒ቪ࣊ࢠࢰ࣭ࡡᵋ⠇㜷㒂 Ṅ㸡Evelyn B. Elegado㸡᭧᰷㍜㞕 㸝໪ኬ㝌㎨࣬ᚺ⏕ⳞᏕ㸞࠘┘Ⓩ࠙㎾ᖳ㸡⣊≟Ⳟࡡࢣࢿ࣑よᯊ࠿㐅ࡳ࡞ࡗࡿ㸡ኣࡂࡡ Hypothetical Protein 㐿ఎᏄ࠿ྜྷᏽࡈࡿ࡙࠷ࡾ㸣ࡆࡿࡼࡡᶭ⬗ࢅよᯊࡌࡾ࡞ࡢ㸡㐿ఎᏄ◒ቪᐁ㥺࠿୘ྊḖ࡚࠵ࡾ㸣㐿ఎᏄ◒ቪᐁ㥺ࡢ㸡┞ྜྷ⤄ᥦ࠻⣌ࢅฺ⏕ࡊ࡙⾔ࢂࡿࡾࡆ࡛࠿ኣ࠷࠿㸡ࡐࡡ◒ቪ⏕ DNA ࡡషᠺ࡞ࡢ㸡々ᩐ᩷∞ࡡࢠ࣭ࣞࢼࣤࢡࡷ㸡fusion PCR ࢅ⏕࠷ࡾ᪁Ἢ㸡ࢹࣚࣤࢪ࣎ࢯࣤࢰ࢟ࣤࢡ࡝࡜࠿࠵ࡾ㸣௑ᅂᠻࠍࡢ Inverse PCR ࢅ⏕࠷㸡ࡈࡼ࡞ GATEWAY ࢨࢪࢷ࣑ࢅฺ⏕ࡌࡾࡆ࡛࡚⠾౼࡞◒ቪ⏕ DNA ࢅషᠺࡌࡾ⣌࡛㸡ࡐࡿ࡞⏕࠷ࡾ࣊ࢠࢰ࣭ࡡᵋ⠇ࢅ⾔ࡖࡒ㸣࠘᪁Ἢ࡛⤎ᯕ࠙Inverse PCR ࡞ࡻࡽ㸡ᵾⓏ㐿ఎᏄࡡ㝼᥃ DNA ࢅቌᖕࡊ㸡࣏࣭࣭࢜㐿ఎᏄࢅᣚࡗ࣊ࢠࢰ࣭࡞㏻⤎ᚃ㸡Inverse PCR ࡞౐⏕ࡊࡒโ㝀㓕⣪࡚㛜⎌ࡌࡾࡆ࡛࡚㸡◒ቪ⏕ DNA ࠿షᠺ࡚ࡀࡾ㸣ࡆࡡ࣊ࢠࢰ࣭ࡢ㸡โ㝀㓕⣪ࢦ࢕ࢹࢅᣚࡒࡍ㸡࡝࠽࠾ࡗ⠾౼࡞ Inverse PCR ᩷∞࠿⤎ྙ࡚ࡀࡾࡆ࡛࠿᭻ࡱࡊ࠷㸣pGEM-Teasy ࣈࣚࢪ࣐ࢺࡡ MCS ࢅ⬲ⴘࡈࡎ㸡ࡐࡆ࡞ pCB1004 ࡡࣀ࢕ࢡ࣏ࣞ࢕ࢨࣤ⪇ᛮ㐿ఎᏄࢅ᣼ථࡊ㸡ࡈࡼ࡞ GATEWAY ⏕ࡡ
attR-CmR-ccdB-attR ᵋ㏸ࢅ㏻⤎ࡌࡾࡆ࡛࡚㸡pDESTR ࢅషᠺࡊࡒ㸣ࡆࡡ࣊ࢠࢰ࣭࡞ࡢ௥⾪Ⓩ࡝โ㝀㓕⣪ࡡࢦ࢕ࢹ࠿࡝ࡂ㸡GATEWAY ࡞ࡻࡽ Inverse PCR ᩷∞ࢅຝ⋙ࡻࡂ᣼ථ࡚ࡀࡾ㸣࣓ࢸ࡛ࣜࡊ࡙ Neurospora crassa ࡡ Frost㐿ఎᏄࡡ◒ቪᐁ㥺ࢅ⾔ࡖࡒ㸣 Frost 㐿ఎᏄࡡ࿔㎰ 3.6kb ࡡ XhoI ᩷∞ࢅ Inverse PCR ࡚ቌᖕࡊ㸡pENTR-D-TOPO
࡞ࢠ࣭ࣞࢼࣤࢡࡊ㸡pDESTR ࡞⤄ᥦ࠻㸡XhoI ࡚ᾐ໩ࡊࡒ㸣ࡆࡿࢅ⏕࠷࡙ N. crassa 㔕⏍ᰬࢅᙟ㈻㌹ᥦࡊࡒ࡛ࡆ
ࢀ㸡ኣࡂࡡ frost ን␏ᰬᵕࡡࢤࣞࢼ࣭࠿ᚋࡼࡿࡒ㸣ࢦࢧࣤよᯊ࡞ࡻࡽ㸡㐿ఎᏄ◒ቪ࠿☔ヾࡈࡿࡒ㸣
Construction of GATEWAY-based gene disruption vectors for filamentous fungi
Ayumi Abe, Evelyn B. Elegado, Teruo Sone
(Lab. Appl. Microbiol., Grad. Sch. Agr., Hokkaido Univ.)

53

P-7
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ DNA ⤄ᥦ࠻ಞᚗ㐿ఎᏄ⩄ࡡよᯊ
Evelyn B. Elegado, 㜷㒂 Ṅ㸡Marites A. Sales㸡ᒷᓧ㜷ᑋ⨶㸡▴஬༐὘* 㸡᭧᰷㍜㞕㸝໪ኬ㝌㎨࣬ᚺ⏕ⳞᏕ, *ඔᇳኬ⌦࣬⏍మโᚒ㸞࠘┘Ⓩ࠙࠷ࡵࡔ⑋Ⳟࡡ⑋ཋᛮ࣭ࣝࢪን␏࡞ࡢ㸡ࢹࣚࣤࢪ࣎ࢯࣤࡡ㌹⛛ࡷኬぞᶅ࡝᯹Ⰵమ්⥽ᠺ➴㸡DNA 㙈ࡡ⤄ᥦ࠻࠿㛭୙ࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾ㸣⣵⬂හ࡚ࡡ DNA ⤄ᥦ࠻ࢅᢰࡖ࡙࠷ࡾࡡࡢ DNA ⤄ᥦ࠻ಞᚗ㐿ఎᏄ⩄࡚࠵ࡾ࠿㸡࠷ࡵࡔ⑋Ⳟ࡞࠽ࡄࡾࡐࡿࡼ㐿ఎᏄ⩄ࡡよᯊࡡሒ࿈ࡢ࡝࠷㸣ࡐࡆ࡚ᠻࠍࡢ࢕ࢾ࠷ࡵࡔ⑋Ⳟ࡞࠽ࡄ
ࡾ DNA ⤄ᥦ࠻ಞᚗ㐿ఎᏄ⩄ࡡࢠ࣭ࣞࢼࣤࢡࢅ⾔࠷㸡DNA ⤄ᥦ࠻ࡡ࠷ࡵࡔ⑋Ⳟ࣭ࣝࢪን␏࡞࠽ࡄࡾᙲ๪ࢅ᪺
ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙よᯊࢅ㐅ࡴ࡙࠷ࡾ㸣࠘᪁Ἢཀྵࡦ⤎ᯕ࠙⤄ᥦ࠻ಞᚗ㐿ఎᏄ⩄ࡡ୯࠾ࡼ㸡┞ྜྷ⤄ᥦ࠻࡞㛭ࢂࡾ㐿ఎᏄ RAD52, RAD54㸡㟸┞ྜྷᮆ❻⤎ྙ
㸝NHEJ㸞࡞㛭ࢂࡾ㐿ఎᏄ MRE11, Ku70, Ku80 ࡡ࠷ࡵࡔ⑋Ⳟ࣓࣌ࣞࢡ㸡Rhm52, Rhm54, Mhm11, Khm70, Khm80
ࢅ,Degenerate PCR ཀྵࡦࢣࢿ࣑ࢸ࣭ࢰ࠾ࡼࡡ PCR ࡞ࡻࡽ㸡᪝ᮇ⏐࢕ࢾ࠷ࡵࡔ⑋Ⳟ Ina168 ᰬࡻࡽཱིᚋࡊࡒ㸣ሲᇱ㒼าよᯊࡡ⤎ᯕ㸡ࡆࡿࡼࡡ࣓࣌ࣞࢡࡢ N. crassa ࡡ࣓࣌ࣞࢡ࡛᭩ࡵ㧏࠷┞ྜྷᛮࢅ♟ࡊࡒ㸣Rhm52 ࡛ Rhm54
ࢅ N. crassa mus-11, mus-25 ን␏ᰬ࡞ᑙථࡊࡒ࡛ࡆࢀ㸡ን␏ᰬࡡ MMS(࣒ࢲ࣒ࣜࢰࣤࢪࣜ࣌ࣤ㓗)វུᛮࢅࡐࡿ
ࡑࡿ┞⿭ࡊ㸡ᶭ⬗ࡌࡾ஥࠿☔ヾࡈࡿࡒ㸣ࡱࡒࢿ࣭ࢧࣤよᯊࡡ⤎ᯕ㸡Rhm52㸡Rhm54 ࡢ MMS㸡UV ࡡን␏″࡞ຊ࠻㸡⇍ࢨࣘࢴࢠ㸝42Υ㸞㸡Methyl Viologen ࡚ࡵࡐࡡ㌹෕ࡡ୕᪴࠿☔ヾࡈࡿ㸡ᐙ୹ࡡᢤᢘᛮཬᚺ࡞ࡻࡾⓆ⌟ࡡቌኬ࠿㉫ࡆࡾྊ⬗ᛮࢅ♟ࡊ࡙࠷ࡾ㸣⌟ᅹ㸡Khm70, Khm80 ࡡ N. crassa ን␏ᰬࡡ┞⿭モ㥺࡛㸡ࡐࡿࡑࡿࡡ㐿ఎᏄࡡ pDESTR ࢅ⏕࠷ࡒ◒ቪᰬࡡషᠺࢅ⾔ࡖ࡙࠷ࡾ㸣
Analyses of recombination al repair genes in the rice blast fungus
Evelyn B. Elegado, Ayumi Abe, Marites A. Sales, Asumi Iwasaki , Chizu Ishii*, Teruo Sone
(Lab. Appl. Micriobiol. Grad. Sch. Agr., Hokkaido Univ. *Saitama Univ.)

P-8࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ௛╌ჹᙟᠺ࡞ᑊࡌࡾ࣏ࢠࣞࣚ࢕ࢺ⣌ᢘ⏍∸㈻ࡡᙫ㡢Ṃᕖ ἖㸡㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ㸞࣏ࢠࣞࣚ࢕ࢺ⣌ᢘ⏍∸㈻ࡢཋᰶ⏍∸ࡡ 50S ࣛ࣍ࢮ࣭࣑࡞⤎ྙࡊ㸡࣋ࣈࢲࢺ㌹⛛ཬᚺࢅ㜴ᐐࡌࡾࡆ࡛࡞ࡻࡽࢰ
ࣤࣂࢠ㈻ྙᠺࢅ㜴ᐐࡌࡾࡆ࡛࡚▩ࡼࡿ࡙࠷ࡾ. ࡊࡒ࠿ࡖ࡙㸡┷ᰶ⏍∸࡞࠽࠷࡙ࡢ࣐ࢹࢤࣤࢺࣛ࢓࡝࡜㸡ཋᰶ⏍∸ᆵࡡࣛ࣍ࢮ࣭࣑ࢅᣚࡗ࢛ࣜ࢝ࢾࣚ࡞ష⏕ࡊ㸡࿣ྺ㜴ᐐ➴࠿࠽ࡆࡾࡆ࡛࠿஢᝷ࡈࡿࡾ. ࡊ࠾ࡊ࡝࠿ࡼ㸡࣓ࢲࣛࣤᵕష⏕㸡IL-8 ➴ࡡࢦ࢕ࢹ࢜࢕ࣤࡡ⏐⏍㜴ᐐష⏕㸡⾉⟮᩺⏍㜴ᐐష⏕࡝࡜ᚉᮮࡡᢘⳞష⏕࡚ࡢㄕ᪺࡚ࡀ
࡝࠷ష⏕࠿ሒ࿈ࡈࡿ࡙࠷ࡾ. ࡐࡆ࡚㸡࣏ࢠࣞࣚ࢕ࢺ⣌ᢘ⏍∸㈻࠿࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ௛╌ჹฦ໩࡞ࡵᙫ㡢ࢅ୙࠻࡝࠷࠾᳠ᏽࢅ⾔࠷㸡࿣ྺࢅ㜴ᐐࡎࡍ࡞ฦ໩㜴ᐐὩᛮࢅ♟ࡌലྡྷ࠿ぜࡼࡿࡒ. ࡱࡒ㸡ཋᰶ⏍∸࡞࠽࠷࡙ࢰࣤ
ࣂࢠྙᠺ㜴ᐐຝᯕ࠿࠵ࡾ௙ࡡᢘ⏍∸㈻࡞ࡗ࠷࡙ࡵྜྷᵕࡡ᳠ᏽࢅ࠽ࡆ࡝ࡖࡒ࡛ࡆࢀ㸡ࢠ࣑ࣞࣚࣆ࢘ࢼࢤ࣭࡚ࣜ
ࡵ௛╌ჹฦ໩ࡡ㜴ᐐ࠿ふᐳࡈࡿࡒ.
The effect of macrolide antibiotics on appressorial development of Magnaporthe gresea.
Osamu Takwkawa,Takashi Kamakura.

(Tokyo Univ.of Sciense)

54

P-9
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡⴝ㠟⣵⬂හ࡚ࡡវ᯹ິឺ࡛㐿ఎᏄⓆ⌟୔⏛ᆀ㈏ྍ࣬㰳⸠៿ୌ㑳࣬㔘᲻ḿᶖ࣬᭯Ờງ࣬ᑈᒱᚥ 㸝㎨ᕝኬ㎨㸞
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ⬂ᏄⓆⰾ㸡௛╌ჹᙟᠺ࠾ࡼᐙ୹࢕ࢾ࡫ࡡ౴ථ࡞⮫ࡾវ᯹ิ᭿㐛⛤࡞ࡗ࠷࡙ࡢኣࡂࡡ▩ぜ࠿ᚋࡼࡿ࡙࠷ࡾ࠿㸡ᐙ୹౴ථᚃࡡິឺ࡞ࡗ࠷࡙ࡢᮅレࡡ㒂ฦ࠿ኣ࠷. EGFP ᖏⓆ⌟ᰬࢅ⏕࠷࡙࢕ࢾⴝ㠟⣵⬂࡫ࡡ౴ථ㸡౴ථⳞ⣊ࡡఘᒈ㸡ⶕᘇࡡᣪິࢅභ↌Ⅴ㢟᚜㙶ୖ࡚ふᐳࡊࡒ࡛ࡆࢀ㸡⮾࿝῕࠷ᣪິ࠿ふᐳࡈࡿࡒ. ౴ථⳞ⣊ࡢ㝼᥃⣵⬂࡫ࡡ౴ථ᫤࡞⣵⬂ባ௛㎾࡚࠷ࡖࡒࢆ⭶₮ᚃ㸡ࡂࡦࡿ⣵ࡱࡖࡒᚃ㸡⣵⬂ባࢅ㈇ථࡊ㸡ࡐࡡ㒂న࡚ࡢ EGFP ࡡ⺧ක࠿ぜࡼࡿ࡝࠷࠾᚜ᘽ࡚࠵ࡖࡒ. ࡐࡡᚃ㸡౴ථⳞ⣊ࡢ඙❻࠿୷ࡲࢅᖈࡦࡒྺჹᵕࡡ⌣≟ᵋ
㏸ࢅ࡛ࡖࡒ. ⤊᫤Ⓩ࡞ふᐳࡌࡾ࡛㸡᥃⛸⣑ 96 ᫤㛣ᚃࡱ࡚ࡡᣪິ࠾ࡼ౴ථⳞ⣊ࡡఘᒈࡢ஦ࡗࡡᵕᘟ࡞ኬื࡚ࡀ
ࡒ. ࡌ࡝ࢂࡔ౴ථ㛜ጙ├ᚃ࠾ࡼふᐳࡈࡿࡾఘᒈ࠿㐔ࡂኯ࠷㸡ฦᯖࢅᩐኣࡂࡵࡗⳞ⣊࡛㸡96 ᫤㛣ᚃ࡞ぜࡼࡿࡒ࡮࡛ࢆ࡜ฦᯖࢅఔࢂࡍ⣵ࡂఘᒈ࠿㏷࠷Ⳟ⣊࡚࠵ࡾ. វ᯹ิ᭿㐛⛤࡞㛭ࢂࡾ FMI1(B19)㐿ఎᏄ 1)ࡡ ORF ୕Ὦ⣑
2.1 kb ࡡ᩷∞ࡡୖὮ࡞ EGFP ࢅ㏻⤎ࡊ㸡ࡐࡡᑙථ಴మࢅⴝ㠟᥃⛸ࡊ࡙ふᐳࡊࡒ࡛ࡆࢀ㸡᥃⛸ 48 ᫤㛣ᚃࡡ๑⩽ࡡ౴ථⳞ⣊࡞ࡢ⺧ක࠿ヾࡴࡼࡿࡒࡵࡡࡡ㸡90 ᫤㛣ᚃࡡ౴ථⳞ⣊࡚ࡢ⺧ක࠿ᾐ኶ࡊ࡙࠷ࡒ. ࡆࡿࡼࡡࡆ࡛ࡢᮇ
Ⳟࡡ౴ථⳞ⣊ࡡິឺ࠿࢕ࢾ⤄⧂හ࡚ን໩ࡊ㸡ࡐࡿ࡛➚ྒࡊ࡙Ⓠ⌟ࡌࡾ㐿ఎᏄࡵวࡽ᭨ࢂࡾࡆ࡛ࢅ♟ြࡊ࡙࠷ࡒࠊ1)Saitoh et al. (2005) 23rd Fungal Genetics Conference program book p137
Dynamic behavior of rice blast fungus expressing EGFP in rice sheath cells and associated genes expression.
Takashi Mitachi, Ken-ichiro Saitoh, Masaki Kanamori, Tsutomu Arie and Tohru Teraoka.
(Tokyo Univ. Agric. & Tech.)P-10࢕ࢾ࠷ࡵࡔ⑋Ⳟ௛╌ჹᙟᠺ㛭㏻㐿ఎᏄ CBP1 ⏐∸ࡡᑻᅹᛮࡡ᳠ゞᒱᮇಆ㍔㸡㰳⸠៿ୌ㑳*㸡᭯Ờ ງ*㸡ᑈᒱ ᚥ*㸡㙂ಲ㧏ᚷ 㸝᮶⌦ኬ⌦ᕝ࣬*㎨ᕝኬ㎨㸞
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ௛╌ჹᙟᠺㄇᑙ࡞㛭୙ࡌࡾ CBP1 ࠿ࢤ࣭ࢺࡌࡾࢰࣤࣂࢠ㈻ࡢ N ᮆ❻ഁ࡞⭯㏩㐛ࢨࢡࢻࣜ㒼า࠿Ꮛᅹࡌࡾࡆ࡛࠾ࡼ㸡CBP1 㐿ఎᏄ⏐∸ࡢ⣵⬂⭯⣌ཧࡢ⣵⬂አ࡫⛛⾔ࡌࡾ࡛஢ῼࡈࡿࡾ㸣ࡱࡒᮇࢰࣤࣂࢠ㈻ࡢ C ᮆ❻ഁ࡞ࢬ࡛ࣛࣤࢪ࢛ࣝࢼࣤ࡞ᐣࢆࡓ㒼าࡵྱࢆ࡚࠽ࡽ㸡ࡆࡡࡻ࠹࡝㒼าࢅᣚࡗࢰࣤࣂࢠ㈻࡞࠽࠷࡙ GPI࢓࣭ࣤ࢜໩࡞ࡻࡖ࡙⣵⬂⭯࡞ᏽ╌ࡊ࣎ࣛ࣋ࣈࢲࢺ㙈ࢅ⣵⬂ባහ࡞ᒈ㛜ࡊ࡙ୌ㒂ࢅ⣵⬂አ࡫㟚ฝࡌࡾౚ࠿ぜ࠷ࡓࡈࡿ࡙࠷ࡾ㸣ࡵࡊ㸡CBP1 㐿ఎᏄ⏐∸࠿ GPI ࢓࣭ࣤ࢜໩ࡈࡿ࡙࠷ࡾ࡝ࡼࡣᮇࢰࣤࣂࢠ㈻ࡡᑻᅹᛮ࡛ష⏕㒂నࢅ≁ᏽࡌࡾ୕࡚ᙁງ࡝ッᣈ࡛࡝ࡾ㸣GPI ࢓࣭ࣤ࢜໩ࡈࡿࡒࢰࣤࣂࢠ㈻ࡢ PI-PLC ฌ⌦࡞ࡻࡖ࡙⬙⫣㓗㙈࡛㐗㞫ࡌࡾࡒࡴ㸡CBP1 ࡡ C ᮆ❻࡛ GFP ࡡ⼝ྙࢰࣤࣂࢠ㈻ࢅᵋᠺⓏ࡞Ⓠ⌟ࡌࡾᰬࢅషฝࡊ㸡ࣈࣞࢹࣈࣚࢪࢹ໩࠽
ࡻࡦࣈࣞࢹࣈࣚࢪࢹࡡ PI-PLC ฌ⌦࡞ࡻࡾ⺧කን໩ࢅ㢟᚜㙶ふᐳ࠽ࡻࡦ᧔ᙫ⏤ാࡡࢸࢩࢰࣜよᯊࢅ⾔࡝࠹ࡆ࡛࡞ࡻࡖ࡙ࡐࡡᑻᅹ࡛ GPI ࢓࣭ࣤ࢜໩ࡡ᭯↋ࢅ᳠ゞࡊࡒ㸣The localization of CBP1 protein of Magnaporthe grisea.
Shunsuke Okamoto,Kenichiro Saitoh,Tsutomu Arie, Toru Teraoka,and Takashi Kamakura.
(Tokyo Univ.of Sciense,Tokyo Univ.of Aguric.&Technol)

55

P-11
࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ Hik1 ࡡ᝗ሒఎ㐡⣌ࡡよᯊ᲻⏛┷⣟ 1,2㸡ᮇᒜ㧏ᖶ 1㸡Ꮻఫ⨶ㄵ 2㸡ᕝ⸠ಆ❮ 1 㸝1 ⌦◂࣬୯ኳ◂ࠉ2 ᮶Ὂኬ࣬ᕝ㸞࠘┘Ⓩ࠙࢕ࢾࡡ᭩㔔さ⑋ཋⳞ࡚࠵ࡾ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡࣀ࢕ࣇࣛࢴࢺᆵࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ Hik1 ࡡ null ን␏మ㸝Δhik1㸞ࡢ㧏⃨ᗐࡡ⢶࡞ᑊࡌࡾ㧏ᾈ㏩ᅸវུᛮࢅ♟ࡊࠉ⑋ཋᛮࡢ㔕⏍ᰬ࡛ንࢂࡼ࡝࠷ࠊୌ᪁ࠉୌ㒂ࡡⅤን␏మ㸝hik1-M16 ᆵን␏మ㸞ࡢ㧏ᾈ㏩ᅸ㉰វུᛮཀྵࡦ⑋ཋᛮࡡ఩ୖࢅ♟ࡌࠊࡆࡡࡻ࠹࡝ᙟ㈻ࢅ♟ࡌཋᅄࢅ᪺ࡼ࠾
࡞ࡌࡾ࡛࡛ࡵ࡞ࠉHik1 ࡡ᝗ሒఎ㐡⣌ࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊ࡙よᯊࡊࡒࠊ࠘᪁Ἢ࣬⤎ᯕ࠙㓕ẍ࠾ࡼࣃࢹࡱ࡚㧏ᾈ㏩ᅸ➴ࡡࢪࢹࣝࢪᚺ➽࡞ࡢ p38 MAP ࢞ࢻ࣭ࢭ࠿㛭୙ࡌࡾࠊ࢕ࢾ࠷ࡵࡔ
⑋Ⳟ࡞࠽࠷࡙ࡢ Osm1 ࠿ࡆࡿ࡞┞ᙔࡌࡾࠊHik1 ࠾ࡼࡡ᝗ሒ࠿ࡆࡡ MAPK ⤊㊪ࢅὩᛮ໩ࡌࡾ࠾࡜࠹࠾ࢅࡲࡾࡒ
ࡴ࡞ࠉΔhik1 ᰬཀྵࡦ hik1-M16 ᰬࢅ㧏ᾈ㏩ᅸฌ⌦ࡊࡒሔྙࡡ Osm1 ࡡὩᛮ໩ࢅよᯊࡊࡒࠊhik1-M16 ᰬ࡚ࡢࠉ㧏
ᾈ㏩ᅸฌ⌦ࡊࡒሔྙࡡ Osm1 ࡡὩᛮ໩ࣝ࣊ࣜࡢ㔕⏍ᰬࡷ Δhik1 ᰬ࡛Ẓ㍉ࡊ࡙఩ୖࡊ࡙࠽ࡽࠉHik1 ࡢ Osm1 ࡡὩᛮ໩࡞రࡼ࠾ࡡ㛭୙ࢅࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡱࡒࠉhik1-M16 ᰬ࡚ࡡ Osm1 ࡡὩᛮ໩ࣝ࣊ࣜࡡ఩ୖࡢ
㧏ᾈ㏩ᅸ㉰វུᛮࡡୌࡗࡡཋᅄ࡚࠵ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ

Analysis of the signal transduction pathway of a histidine kinase Hik1 of the rice blast fungus
Masumi Morita1,2, Takayuki Motoyama1, Ron Usami2, Toshiaki Kudo1(1Discovery Res. Institute, RIKEN, 2Fac. Technol., Univ. Toyo)

P-12࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ஦ᠺฦ᝗ሒఎ㐡⣌ࡡ⎌ሾᚺ➽࡫ࡡ㛭୙ࡡよᯊྡྷᆊ⏜㈏㸡ᮇᒜ㧏ᖶ㸡ᕝ⸠ಆ❮㸝⌦◂࣬୯ኳ◂㸞
࠘┘Ⓩ࠙஦ᠺฦ᝗ሒఎ㐡⣌ࡢࣁࢠࢷࣛ࢓࠾ࡼⳞ㢦ࡷ᳔∸࡝࡜࡞࠽࠷࡙⎌ሾᚺ➽࡝࡜࡞㛭୙ࡌࡾࠊ࢕ࢾࡡ᭩㔔さ⑋ཋⳞ࡚࠵ࡾ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡࢣࢿ࣑୯࡞ࡢ஦ᠺฦ᝗ሒఎ㐡⣌ᅄᏄ࡛ࡊ࡙ࠉ10 ⛸ࡡ㸝᥆ᏽ㸞ࣀ࢕ࣇࣛࢴࢺ
ᆵࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭࠉ1 ⛸ࡡ᥆ᏽྱࣃࢪࢲࢩࣤࣛࣤ㓗㌹⛛ࢰࣤࣂࢠ㈻ࠉ3 ⛸ࡡ᥆ᏽࣝࢪ࣎ࣤࢪ࣭ࣝ࢟ࣖࣝࢰ
࣭࠿Ꮛᅹࡌࡾࠊ஦ᠺฦ᝗ሒఎ㐡⣌ᅄᏄ࡞Ἰ┘ࡊࠉ࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ஦ᠺฦ᝗ሒఎ㐡⣌ࡡ⎌ሾᚺ➽ࡷ⑋ཋᛮ࡫
ࡡ㛭୙ࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡌࡾࠊ࠘᪁Ἢ࣬⤎ᯕ࠙࢕ࢾ࠷ࡵࡔ⑋Ⳟࡡ஦ᠺฦ᝗ሒఎ㐡⣌ࡡᶭ⬗ࢅよᯊࡌࡾࡒࡴࠉ3 ࡗࡡ᥆ᏽࣝࢪ࣎ࣤࢪࣝ࢟ࣖࣝ
࣭ࢰ࣭㐿ఎᏄ㸝MgSSK1ࠉMgSKN7ࠉMgRIM15㸞ࡡ㐿ఎᏄ◒ቪమࢅషᠺࡊࡒࠊ3 ⛸ࡡ㐿ఎᏄ◒ቪమ࡛ࡵ㏳ᖏࡡ
ᇰᆀ࡚ࡡ⏍⫩㏷ᗐࡢ㔕⏍ᰬ࡛ྜྷᵕࡓࡖࡒࠊMgSSK1 㐿ఎᏄ◒ቪᰬ࡚ࡢ㢟ⴥ࡝㧏ᾈ㏩ᅸវུᛮࢅ♟ࡊࡒࠊୌ᪁ࠉ
㓗໩ࢪࢹࣝࢪវུᛮࡢ࡜ࡡ㐿ఎᏄ◒ቪమ࡚ࡵ㔕⏍ᰬ࡛ྜྷ⛤ᗐࡓࡖࡒࠊ

Functional analysis of the two-component signal transduction system of the rice blast fungus in response toenvironmental signals
Yuki Mukouzaka, Takayuki Motoyama, Toshiaki Kudo
(Discovery Res. Institute, RIKEN)

56

P-13
Neurospora crassa ࡡᾈ㏩ᅸ OS ࢨࢡࢻࣜఎ㐡⤊㊪ࡡୖὮ࡚โᚒࡈࡿࡾ㐿ఎᏄࡡ ྜྷᏽ㔕ཾⲸᯖᏄ㸡ᆊ㔕┷ᖲ, ᕰᕖீኯ, ᮄᮟ┷ 1, ᒜཾຩ 2, ⸠ᮟ┷㸝᮶Ὂኬ࣬⏍࿤, 1 ⌦◂, 2 ᮶Ὂኬ࣬ᕝ㸞࢓࢜ࣂࣤ࢜ࣄࡡᾈ㏩ᅸ OS ࢨࢡࢻࣜఎ㐡⤊㊪࡛㓕ẍࡡᾈ㏩ᅸࢨࢡࢻࣜఎ㐡 HOG ⤊㊪ࡢࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ
࡛ MAP ࢞ࢻ࣭ࢭ࢜ࢪࢢ࣭ࢺ࠾ࡼᵋᠺࡈࡿ࡙࠽ࡽࠉos-2 MAP ࢞ࢻ࣭ࢭࡢ HOG1 ࢅᶭ⬗Ⓩ࡞┞⿭ࡌࡾࡆ࡛࠿ሒ
࿈ࡈࡿ࡙࠷ࡾࠊHOG ⤊㊪ࡡୖὮ࡚ࡢࠉCTT1㸝Catalase㸞ࡷ GPD1, GPP1㸝glycerol ྙᠺ㓕⣪㸞, HSP12㸝heat shockprotein㸞, FBP1㸝⢶᩺⏍㛭㏻㓕⣪㸞࡝࡜ࡡ㐿ఎᏄ࠿โᚒࡈࡿ࡙࠷ࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉOS ⤊㊪ࡡୖὮ࡚โᚒࡈࡿ࡙࠷ࡾ㐿ఎᏄ࡞ࡗ࠷࡙ࡢ࡮࡛ࢆ࡜ሒ࿈࠿࡝࠷ࠊࡐࡆ࡚ࠉHOG ⤊㊪ࡡୖὮ࡚โᚒࡈࡿ࡙࠷ࡾ
㐿ఎᏄࡡ࢛ࣜࢮࣞࢡࢅ࢓࢜ࣂࣤ࢜ࣄࡡࢣࢿ࣑᝗ሒࡻࡽ᳠⣬ࡊࠉ㐿ఎᏄࡡⓆ⌟ንິࢅࣛ࢓ࣜࢰ࢕࣑ PCR Ἢ࡞ࡻ
ࡽよᯊࡊࡒࠊ࡝࠽ࠉOS ⤊㊪ࡡୖὮ࡚โᚒࡈࡿࡾ㐿ఎᏄࡡ≁ᏽࡢࠉ1㸞ᾈ㏩ᅸฌ⌦࠽ࡻࡦ OS ⤊㊪࡞ష⏕ࡌࡾṽⳞ๠ Fludioxonil ฌ⌦ࡡ୦᪁࡞ࡻࡽⓆ⌟࠿ንິࡌࡾࠊ2㸞os-2 ን␏ᰬ࡚ࡢࢪࢹࣝࢪฌ⌦࡞ࡻࡾⓆ⌟ንິ࠿ᾐ኶
ࡌࡾࠊࡆ࡛ࢅᣞᵾ࡛ࡊࡒࠊࡐࡡ⤎ᯕࠉNcCTT1 ཀྵࡦ NcFBP1 ࡢ 1㸞2㸞࡛ࡵ࡞ぜࡼࡿࠉ㓕ẍ࡛ྜྷᵕ࡞ OS ⤊㊪ࡡୖὮ࡚โᚒࡈࡿ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡊ࠾ࡊࠉNcGPD1, NcGPP ࡷ HSP ࡡᾈ㏩ᅸࠉṽⳞ๠ฌ⌦࡞ࡻࡾㄇᑙࡢヾࡴࡼࡿ࡝࠾ࡖࡒࠊࡆࡡࡆ࡛ࡻࡽࠉHOG ⤊㊪࡛ OS ⤊㊪ࡡୖὮ࡚โᚒࡈࡿ࡙࠷ࡾ㐿ఎᏄ⩄࡞ࡢ㐢࠷࠿࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊHOG ⤊㊪࡛ྜྷᵕ࡞ࠉOS ⤊㊪ࡡὩᛮ໩ࡢ⣵⬂හࢡࣛࢬ࣭ࣞࣜ㔖ࢅቌຊࡈࡎࡾ
࠿ࠉ࢓࢜ࣂࣤ࢜ࣄ࡚ࡢࢡࣛࢬ࣭ࣞࣜྙᠺ㓕⣪࡛ࡊ࡙ࠉNcGPD1, NcGPP ࡚ࡢ࡝ࡂࠉNcGLD ࠿ os-2 ౪ᏋⓏ࡞Ⓠ⌟โᚒࡈࡿ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡈࡼ࡞ࠉOS ⤊㊪ࡡୖὮ࡚โᚒࡈࡿ࡙࠷ࡾࡆࡿࡼࡡ㐿ఎᏄࡡⓆ⌟࡞ࡗ࠷࡙ࠉ㓗໩Ⓩࢪࢹࣝࢪࡷࣃ࣭ࢹࢨࣘࢴࢠ࡝࡜ࡡ྘⛸ࢪࢹࣝࢪᚺ➽࡞ࡗ࠷࡙ࡵよᯊࡊࡒࡡ࡚ሒ࿈ࡌࡾࠊ
Neurospora genes induced by osmotic stress and fludioxonil under control of the os-2 MAP kinase.
Rieko Noguchi, Shinpei Banno, Ryouta Ichikawa, Makoto Kimura1, Isamu Yamaguchi2, Makoto Fujimura(Toyo Univ. Grad. Life Sciences, 1RIKEN. Dis. Res. Inst, 2 Toyo Univ. Fac. Engineering)

P-14࢓࢜ࣂࣤ࢜ࣄࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ-MAP ࢞ࢻ࣭ࢭ⤊㊪࡛ cAMP-PKA ⤊㊪ࡡࢠࣞࢪࢹ࣭ࢠῳ㑌⟿Ꮔ㸦㸡ሲ⃕࠵ࡍࡈ㸦ࠉᆊ㔕┷ᖲ 1㸡ᮄᮟ┷ 2㸡⸠ᮟ┷ 1 㸝㸦᮶Ὂኬ࣬⏍࿤㸡㸧⌦◂࣬୯ኳ◂㸞࢓࢜ࣂࣤ࢜ࣄࡡ cAMP-PKA ⤊㊪ࡢฦ⏍Ꮔᙟᠺ࣬Ẵ୯Ⳟ⣊ᙟᠺࡡโᚒ࡞㛭୙ࡊ࡙࠷ࡾࠊᜇᖏⓏὩᛮ໩ን␏࡛᥆
ᏽࡈࡿࡾㄢ⟿ࢦࣇࣗࢼࢴࢹ hah ᰬࡢẴ୯Ⳟ⣊ࡡஸ㐅࡛ฦ⏍ᏄᙟᠺࡡḖ኶࠿ヾࡴࡼࡿࠉྜྷ⤊㊪ࡡ㐵᩷ን␏࡛᥆
ᏽࡈࡿࡾり፳ࣗࢼࢴࢹ pkac-1 ᰬࡢฦ⏍Ꮔᙟᠺ࠿ᙁࡂஸ㐅ࡈࡿ colonial ࡝⏍⫩ࢅࡌࡾࠊୌ᪁ࠉUstilago maydis
࡚ࡢ cAMP-PKA ⤊㊪ࡡን␏ᰬࡡ࡝࠾࡞ࠉᾈ㏩ᅸវུᛮ࡛ iprodione ⪇ᛮࢅ♟ࡌౚ࠿ሒ࿈ࡈࡿ࡙࠽ࡽࠉ࢓࢜ࣂ
ࣤ࢜ࣄࡡ os-1 ࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ-MAP ࢞ࢻ࣭ࢭ(OS)⤊㊪ࡡን␏ᰬࡡᙟ㈻࡛㓖జࡊ࡙࠷ࡾࠊࡐࡆ࡚ࠉ࢓࢜ࣂ
ࣤ࢜ࣄࡡ cAMP-PKA ⤊㊪࡛ᾈ㏩ᅸᚺ➽࡛ࡡ㛭㏻ࢅㄢ࡬ࡒࠊhah ࠽ࡻࡦ pkac-1 ᰬࡡᾈ㏩ᅸࡷ iprodione ࡞ᑊࡌࡾវུᛮࡢࠉ㔕⏍ᰬ࡛㢟ⴥ࡝ᕣࡢヾࡴࡼࡿ࡝࠾ࡖࡒࠊࡱࡒࠉ஦㔔ን␏ᰬ os-1/pkac-1 ࡛ os-1/hah ࡡᙟឺࡢࠉᇱ
ᮇⓏ࡞ࡢ os-1 ን␏࡞ࡻࡾᙫ㡢ࢅུࡄࡍࡐࡿࡑࡿ pkac-1 ࡛ hah ࡡᙟឺࢅ♟ࡊࡒࠊࡊ࠾ࡊࠉ
pkac-1/os-1 ᰬ࡚ࡢ os-1
ᰬࡡࡵࡗᾈ㏩ᅸវུᛮ࠿࡮࡯Ᏸධ࡞ᢒโࡈࡿࠉࡳࡊࢀ 2㸚ࡡ NaCl Ꮛᅹୖ࡚⏍⫩࠿ಀ㐅ࡈࡿࡾࡆ࡛࠿ึ᪺ࡊࡒࠊ
㏣࡞ࠉhah/os-1 ᰬ࡚ࡢࠉos-1 ᰬࡻࡽࡈࡼ࡞㧏࠷ᾈ㏩ᅸវུᛮࢅ♟ࡊࡒࠊࡆࡿࡼࡡࡆ࡛࠾ࡼࠉᾈ㏩ᅸᚺ➽࡞ᑊ
ࡊ࡙ࠉcAMP-PKA ⤊㊪࡛ OS ⤊㊪㛣࡞ࡢࢠࣞࢪࢹ࣭ࢠ࠿࠵ࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊ

Cross talk between histidine kinase-MAP kinase pathway and cAMP-PKA pathway in Neurospora crassa
Setsuko Watanabe1, Azusa Shiozawa㸦, Shinpei Banno1, Makoto Kimura2, and Makoto Fujimura1(1Fac.of Life Sci.,Toyo Univ.,2Env.Mol.Biol.,RIKEN,)

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N. crassa ࡡ々ᩐࡡࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ࠿ᾈ ㏩ᅸᚺ➽࡞㛭୙ࡌࡾሲ⃕࠵ࡍࡈ 1㸡ᒱ⏛᫥౅ 1㸡ⴘྙ์ᖶ 2㸡ᑈ஬᪺✉ 1 㸡ᮄᮟ┷ 2㸡⸠ᮟ┷ 1 㸝㸦᮶Ὂኬ࣬⏍࿤㸡㸧⌦◂࣬୯ኳ◂㸞ࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ(HK)ࡢࠉฝⰾ㓕ẍ࡚ࡢ 1 ⛸㢦(Sln1)ࡡࡲ࡚࠵ࡾ࠿ࠉ⣊≟Ⳟ࡞ࡢኣᩐࡡ㐿ఎᏄ࠿Ꮛᅹࡌࡾࠊ
ࡊ࠾ࡊࠉࡆࡿࡼࡡᶭ⬗࡞ࡗ࠷࡙ࡢ os-1 ᵕ㐿ఎᏄ࠿ᾈ㏩ᅸᚺ➽࡛ṽⳞ๠ fludioxonil ⪇ᛮ࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛௧
አࠉ࡮࡛ࢆ࡜᪺ࡼ࠾࡞࡝ࡖ࡙࠷࡝࠷ࠊࡐࡆ࡚ࠉ⣊≟Ⳟࡡ HK ࡡᶭ⬗ࢅよ᪺ࡌࡾࡒࡴ࡞ࠉ࢓࢜ࣂࣤ࢜ࣄࡡࢣࢿ
࣑࡞Ꮛᅹࡌࡾ 11 ⛸㢦ࡡ HK 㐿ఎᏄࡡ྘◒ቪᰬࡡషᠺࢅ࠽ࡆ࡝ࡖ࡙࠷ࡾࠊ⌟ᅹࡱ࡚࡞ࠉNcFOS1 ࢅ㝎ࡂ 10 ⛸㢦ࡡ HK 㐿ఎᏄ◒ቪᰬࡡ༟㞫࡞ᠺຉࡊࡒࠊࡊ࠾ࡊࠉᚋࡼࡿࡒ྘◒ቪᰬ(os-1 ን␏ᰬࢅ㝎ࡂ)ࡢࠉᾈ㏩ᅸវུᛮࠉṽⳞ๠វུᛮ࠽ࡻࡦᙟឺᙟᠺ࡝࡜࡞㛭ࡊ࡙ࠉ㔕⏍ᰬ࡛㢟ⴥ࡝ᙟ㈻ࡡᕣࡢヾࡴࡼࡿࡍࠉ྘ HK ࡡᶭ⬗ࢅ≁ᏽࡌࡾ࡞⮫ࡼ࡝࠾ࡖࡒࠊࡐࡆ࡚ࠉHK 㐿ఎᏄ㛣ࡡᶭ⬗㔔々ࡡྊ⬗ᛮࢅ᳠ッࡌࡾࡒࡴࠉኣ㔔◒ቪᰬࢅషᠺࡊࡐࡡᙟ㈻ࢅ
ㄢ࡬ࡒࠊฝⰾ㓕ẍ࡚ࡢ SLN1㸝ᾈ㏩ᅸࢬࣤࢦ࣭㸞ࡡ◒ቪᰬࡢ lethal ࡚࠵ࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾ࠿ࠉ࢓࢜ࣂࣤ࢜
ࣄࡡ NcSLN ◒ቪᰬࡢࠉAspergillus ࡡሔྙ࡛ྜྷᵕ࡞㢟ⴥ࡝ᙟ㈻ࢅ♟ࡈ࡝࠾ࡖࡒࠊࡊ࠾ࡊࠉncsln/os-1 ஦㔔ን␏ᰬ
ࡢࠉos-1 ༟≺ᰬࡻࡽࡵ㧏࠷ᾈ㏩ᅸវུᛮࢅ♟ࡊࡒࠊ㏣࡞ࠉSer/Thr ࢞ࢻ࣭ࢭࢺ࣒࢕ࣤࢅࡵࡗ NcHK3.79 ◒ቪᰬࡢࠉos-1 ᰬࡡᾈ㏩ᅸវུᛮࢅᢒโࡊࡒࠊ࡝࠽ࠉncsln/os-1 ஦㔔ን␏ᰬࡢ os-1 ᰬ࡛ྜྷᵕ࡞ⷾ๠㧏ᗐ⪇ᛮ࡚࠵ࡖࡒ
࠿ࠉnchk3.79/os-1 ஦㔔ን␏ᰬࡢ os-1 ᰬࡻࡽࡵⷾ๠វུᛮࢅ♟ࡊࡒࠊࡆࡿࡼࡡࡆ࡛࠾ࡼࠉ々ᩐࡡ HK ࠿ᾈ㏩ᅸᚺ➽࡞㛭୙ࡊ࡙࠽ࡽࠉNcSlnp ࡛ Os1p ࡡ㛣࡞ࡢḿࡡ┞பష⏕࠿ࠉ㏣࡞ NcHK3.79p ࡛ Os1p ࡡ㛣࡞ࡢㇿࡡ┞பష⏕࠿࠵ࡾ࡛⩻࠻ࡼࡿࡒࠊ
Three histidine kinases involved in osmoregulation in Neurospora crassa
Azusa Shiozawa1, Akiyoshi Okada1, Noriyuki Ochiai2, Akiho Terai1, Makoto Kimura2, and Makoto Fujimura1(1Fac.of Life Sci., Toyo Univ.㸡2Env.Mol.Biol., RIKEN,)

P-16
Cryptococcus neoformans ࡡࣀ࢕ࣇࣛࢴࢺᆵ Histidine kinase ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ CnNik1ࡡᶭ⬗よᯊΰỀපᚠ 1)㸡ῳ㎰ူ 1)㸡ஞ஬ගᙢ 1)㸡Drivinya Antra1)㸡ྚぜၤ 2)㸡⏛୯༐ᑖ 2)㸡ᕖᮇ㐅 1)㸝1)༐ⴝኬ࣬┷Ⳟࢬ㸡2)ாኬ㝌࣬㎨㸞
Histidine kinase㸝HK㸞ࡢኣࡂࡡ┷Ⳟ࡚ࣆ࢘ࢼࣜࣅ࣭ࣞࣜ⣌ᢘ┷Ⳟ๠࡞ᑊࡌࡾវུᛮࡷᾈ㏩ᅸㄢ⟿࡞㛭ࢂࡾ
MAPK ࢨࢡࢻࣜఎ㐡⤊㊪ࢅโᚒࡌࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࢰࣤࣂࢠ㈻ࣛࣤ㓗໩㓕⣪࡚࠵ࡾࠊ௑ᅂ㸡ࣃࢹ⑋ཋ┷Ⳟ Cryptococcus neoformans ࡡ HK ࢅࢤ࣭ࢺࡌࡾ࡛᥆ᏽࡈࡿࡾ㐿ఎᏄ CnNIK1 ࢅࢠ࣭ࣞࢼࣤࢡࡊ㸡ᵋ㏸よᯊࢅ⾔ࡖࡒࠊሲᇱ㒼า㸡࢓࣐ࢿ㓗㒼าࢅ⏕࠷ࡒ┞ྜྷ᳠⣬ࡡ⤎ᯕ㸡࢓࢜ࣂࣤ࢜ࣄ㸡࢕ࢾ࠷ࡵࡔ⑋Ⳟࢅࡢࡋࡴ࡛ࡌࡾ
┷Ⳟ㢦ࡡ HK 㐿ఎᏄࡡὩᛮ㒂నࢅ୯ᚨ࡞㧏ࡂಕᏋࡈࡿ࡙࠷ࡒࠊB4500 ⏜ᮮࡡᰜ㣬さịᛮⳞᰬ TAD1 ࢅ⏕࠷࡙㸡ᰜ㣬さịᛮ࣏࣭࣭࢜㐿ఎᏄ࡛ CnNIK1 㐿ఎᏄ࡛ࡡ⨠ᥦ࡞ࡻࡾ㐿ఎᏄ◒ቪࢅ⾔࠷㸡ᙟ㈻㌹ᥦᰬ TAD12 ࢅཱིᚋࡊࡒࠊࣆ࢘ࢼࣜࣅ࣭ࣞࣜ⣌ᢘ┷Ⳟ๠ࣆࣜࢩ࢛࢞ࢮࢼࣜ࡞ᑊࡌࡾវུᛮ࡞ࡗ࠷࡙Ẓ㍉ࡊࡒ࡛ࡆࢀ㸡YPD ᾦమᇰᆀࢅ⏕࠷ࡒሔྙࡡ㔕⏍ᆵᰬ࡞ᑊࡌࡾ MIC ࡢ 3ȣg/ml ࡚࠵ࡖࡒࡡ࡞ᑊࡊ࡙㸡TAD12 ࡚ࡢ MIC ೋ࠿ 48ȣg/ml ࡻࡽࡵኬࡀࡂ㸡ᮇⳞࡡ CnNIK1 㐿ఎᏄࡢ௙ࡡ┷Ⳟ㢦࡛ྜྷᵕ࡞ࣆ࢘ࢼࣜࣅ࣭ࣞࣜ⣌ᢘ┷Ⳟ๠࡞ᑊࡌࡾវུᛮ࡞㛭୙ࡊ࡙࠷ࡒࠊୌ᪁㸡㧏ᾈ㏩ᅸ᮪௲ୖ࡚ࡡ⏍⫩ࡢ㔕⏍ᆵᰬ࡛ᕣ␏࠿ぜࡼࡿ࡝࠾ࡖࡒࠊᾈ㏩ᅸㄢ⟿ࢅโᚒࡌࡾ MAPK࡚࠵ࡾ Hog1 ࡡࣛࣤ㓗໩≟ឺࢅẒ㍉ࡊࡒ࡛ࡆࢀ㸡㔕⏍ᆵᰬࡻࡽ఩ࣝ࣊ࣜ࡝ࣛࣤ㓗໩࠿☔ヾࡈࡿࡒࠊ⌟ᅹ㸡ິ∸࣓ࢸࣜࢅ⏕࠷ࡒវ᯹ᐁ㥺ࢅ⾔࠷㸡ᮇ㐿ఎᏄࡡ⑋ཋᛮ࡫ࡡ㛭୙ࢅ᳠ッ୯࡚࠵ࡾࠊ
Functional analysis of Cryptococcus neoformans histidine kinase gene CnNIK1.
Kiminori Shimizu1), Akira Watanabe1), Katsuhiko Kamei1), Antra Drivinya1), Akira Yoshimi2), Chihiro Tanaka2),
Susumu Kawamoto1) (1)RC-PFMT, Chiba Univ., 2)Grad. Sch. Agr., Kyoto Univ.)

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Aspergillus nidulans ヒスチジンキナーゼ遺伝子 nikA、phkA、phkB の機能解析ᮿᯐⰃ༡ࠉᒜᓧࡹ࠾ࡽࠉ୷஬ῗୌ᭹ࠉຊ⸠㞖ኃࠉᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞[┘Ⓩ] ⣊≟Ⳟ Aspergillus nidulans ࡞ࡢࠉHis-Asp ࣛࣤ㓗࣭ࣛࣝᆵ᝗ሒఎ㐡ᶭᵋ࡞㛭୙ࡌࡾ hybrid ᆵ histidine
kinase ( HK )ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ࠿ 15 ⛸Ꮛᅹࡌࡾࠊᠻࠍࡢࡆࡿࡼ HK 㐿ఎᏄࡡᶭ⬗よ᪺ࢅ┘Ⓩ࡛ࡊ࡙ࠉ⥑⨮Ⓩ࡝◒ቪᰬࡡష⿿ࢅ⾔ࡖ࡙࠷ࡾࠊ௑ᅂࡢࠉNeurospora crassa nik-1/os-1 ࡡ࢛ࣜࢮࣞࢡ࡚࠵ࡾ nikAࠉ୩ࡦ࡞
Shizosaccharomyces pombe phk1/mak2, phk2/mak3 ࡡ࢛ࣜࢮࣞࢡ phkAࠉphk3/mak1 ࡡ࢛ࣜࢮࣞࢡ phkBࠉࡐࡿࡑࡿ
࡞ࡗ࠷࡙㐿ఎᏄ◒ቪࡡᙫ㡢ࢅよᯊࡊࡒࠊ[⤎ᯕ] 1. nikA ◒ቪᰬ N. crassa ࡡ▩ぜ࠾ࡼࠉA. nidulans nikA ◒ቪᰬࡢ㧏ᾈ㏩ᅸវུᛮ࡞࡝ࡾ࡛஢᝷ࡈࡿࡒࠊ
ࡊ࠾ࡊࠉnikA ◒ቪᰬࡢ㧏ᾈ㏩ᅸវུᛮࢅ♟ࡈࡍࠉࡳࡊࢀ఩ᾈ㏩ᅸ᮪௲ୖ࡞࠽࠷࡙⏍⫩࡛↋ᛮ⬂Ꮔᙟᠺ࠿ⴥࡊ
ࡂ㐔ᘇࡊࡒࠊࡱࡒࠉN. crassa nik-1 ◒ቪᰬࡢࠉ⣊≟Ⳟ≁␏Ⓩ࡝㎨ⷾࠉ࢕ࣈࣞࢩ࢛ࣤࠉࣆࣜࢩ࢛࢞ࢮࢼࣜ࡞ᑊࡊ
࡙⪇ᛮࢅ♟ࡌ࠿ࠉnikA ◒ቪᰬࡢࡆࡿࡼⷾ๠Ꮛᅹୖ࡚ࠉ㟸ᏋᅹୖࡻࡽⰃይ࡝⏍⫩ࢅ♟ࡊࡒࠊࢡࣛࢬ࣭ࣞࣜ௥ㅨ
㛭㏻㐿ఎᏄࡡⓆ⌟࡞୙࠻ࡾ nikA ◒ቪࡡᙫ㡢࡞ࡗ࠷࡙ࡢ⌟ᅹよᯊࡊ࡙࠷ࡾࠊ
2. phkAࠉphkB ◒ቪᰬ S. pombe ࡞࠽࠷࡙ࠉHK 㐿ఎᏄࡢ㓗໩ࢪࢹࣝࢪᚺ➽ࡷ᭯ᛮ⏍Ṣ࡞㛭୙ࡊ࡙࠷ࡾ࠿ࠉ྘
HK 㐿ఎᏄࡡ༟≺◒ቪ࡚ࡢࡆࡿࡼࡡ⾪⌟ᆵ࡞ᑊࡌࡾ㢟ⴥ࡝ᙫ㡢ࡢぜࡼࡿ࡝࠷ࠊྜྷᵕ࡞ࠉA. nidulans phkAࠉphkB◒ቪᰬࡵᙟឺ୕㢟ⴥ࡝⾪⌟ᆵࢅ♟ࡈ࡝࠾ࡖࡒࠊ
Physiological roles of histidine kinase genes, nikA, phkA and phkB, in Aspergillus nidulans.
Yoshihiro Matsubayashi, Yukari Yamazaki, Jyunichiro Marui, Masashi Kato, Tetsuo Kobayashi( Grad. Sch. of Bioagricultural Sciences, Nagoya Univ.)

P-18⣊≟Ⳟࡡ Hog1 ᆵ MAPK ࡡὩᛮࡢࢡ࣭ࣜࣈϪࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ࡞ࡻࡖ࡙โ ᚒࡈ ࡿࡾྚぜၤ 1 㸡ᑚᓞᾇᖲ 2,3㸡㧏㔕⩇Ꮟ 2 㸡⏛୯༐ᑖ 1 㸝ாኬ㝌㎨࣬1 ᆀ⎌⛁࣬2 ᚺ⏍∸㸡3 ⌟࣬ࢸ࣭ࣖࢠኬ㸞 ࢹ࣓ࢗࣞࢤࢨࡇࡱⴝᯜ⑋Ⳟࡡࢡ࣭ࣜࣈϪࣃࢪࢲࢩࣤ࢞ࢻ࣭ࢭ㸝HKϪ㸞㐿ఎᏄ Dic1 Ḗ᥾ᰬࡢ㸡ࢩ࢜ࣜ࣍࢞
ࢨ࢕࣐ࢺ๠࡝ࡼࡦ࡞ࣆ࢘ࢼࣜࣅ࣭ࣞࣜ๠㧏ᗐ⪇ᛮࢅ♟ࡊ㸡ᾈ㏩ᅸវུᛮࢅెࡎᣚࡗ㸣ࡱࡒ㸡ᩐ⛸ࡡ⣊≟Ⳟ࡞࠽࠷࡙㸡ࡆࡿࡼࡡṽⳞ๠࠿ Hog1 ᆵ MAPK ࢅ␏ᖏὩᛮ໩ࡈࡎࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖ࡙࠷ࡾ㸣 ᠻࠍࡢ㸡HKϪ
࡛ HOG ⤊㊪࡛ࡡ㛭౿ࢅ᪺ࡼ࠾࡞ࡌࡾࡒࡴ㸡ࢹ࣓ࢗࣞࢤࢨࡇࡱⴝᯜ⑋Ⳟࡡ㔕⏍ᆵᰬ࡝ࡼࡦ࡞ Dic1 Ḗ᥾ᰬࢅ౩モࡊ࡙㸡ᮇⳞࡡ Hog1 ᆵ MAPK㸝BmHOG1㸞ࡡࣛࣤ㓗໩ᵕᘟࢅよᯊࡊࡒ㸣ࡱࡍ㸡Ẓ㍉Ⓩ఩࠷ᾈ㏩ᅸࢪࢹࣝࢪ
࠵ࡾ࠷ࡢ఩⃨ᗐⷾ๠ฌ⌦࡚ࡢ㸡㔕⏍ᆵᰬ࡛Ẓ㍉ࡊ࡙ Dic1 Ḗ᥾ᰬ࡞࠽ࡄࡾ BmHOG1 ࡡࣛࣤ㓗໩ࡢ㢟ⴥ࡞఩ୖࡊࡒ㸣ࡆࡡࡆ࡛࠾ࡼ㸡DIC1 ࡢ BmHOG1 ࡡὩᛮࢅ࣎ࢩࢷ࢔ࣇ࡞โᚒࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒ㸣ୌ᪁㸡㧏ᾈ㏩ᅸ᮪௲࡝ࡼࡦ࡞㧏⃨ᗐࡡⷾ๠ฌ⌦࡚ࡢ㸡Dic1 Ḗ᥾ᰬ࡞࠽࠷࡙ࡵ᪺☔࡝ BmHOG1 ࡡࣛࣤ㓗໩࠿ヾࡴࡼࡿࡒ㸣ࡊࡒ࠿ࡖ࡙㸡ᮇⳞ࡞ࡢ DIC1 ௧አ࡞ࡵ BmHOG1 ࡡὩᛮ໩ࢅโᚒࡌࡾᅄᏄ࠿Ꮛᅹࡌࡾࡆ࡛࠿♟ြࡈࡿࡒ㸣ࡈࡼ࡞㸡࢓࢜ࣂࣤ࢜ࣄࡡ HKϪ࡚࠵ࡾ NIK1㸝OS-1)࡛ Hog1 ᆵ MAPK ࡚࠵ࡾ OS-2 ࡡ㛣࡞ࡵ㢦జࡊࡒ㛭౿࠿ヾࡴࡼࡿࡒࡆ࡛࠾ࡼ㸡ୌ⯙࡞⣊≟Ⳟࡡ Hog1 ᆵ MAPK ࡡὩᛮࡢ㸡HKϪ࡞ࡻࡖ࡙โᚒࡈࡿࡾ࡛⩻࠻ࡼࡿࡒ㸣
Hog1-type MAPK activation is regulated by group III histidine kinase in filamentous fungi.
Akira Yoshimi1, Kaihei Kojima2,3, Yoshitaka Takano2, Chihiro Tanaka1(1,2Grad. Sch. Agri., Kyoto Univ., 3Present address: Duke Univ.)

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࢓࢜ࣂࣤ࢜ࣄࡡࢨࢡࢻࣜ㐿ఎᏄ ncSCD1ࠉncSCD2 ࡡよᯊᕖᮟ┷ᚷ㸡ୌ▴᫓ᙢ 㸝᮶Ὂኬ࣬⏍࿤⛁Ꮥ㒂㸞
S.pombe ࡞࠽ࡄࡾ Cdc42 ౪ᏋⓏࢨࢡࢻࣜఎ㐡⤊㊪ࡢ⣵⬂ࡡᴗᛮᠺ㛏ࡷ࢓ࢠࢲࣤᵋ㏸ᙟᠺ࡝࡜ࠉ⣵⬂ࡡᙟឺᙟᠺࢅ⥌ᣚࡊ࡙࠽ࡽࠉGDP/GTP ஹᥦᅄᏄ࡚࠵ࡾ Scd1 ࡛ࠉCdc42 ⤎ྙ⺦Ⓣ㈻࡚࠵ࡾ Scd2 ࠿ Cdc42 ࢰࣤࣂࢠ㈻ࡡㄢ⟿࡞㛭୙ࡊ࡙࠷ࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊ
࢓࢜ࣂࣤ࢜ࣄ࡞࠽࠷࡙ࡵ Scd1ࠉScd2 ࢓࣐ࢿ㓗㒼า࡛┞ྜྷᛮࢅ♟ࡌ㐿ఎᏄ࠿ࢣࢿ࣑ࢸ࣭ࢰ࣭࣊ࢪ୕࡞Ꮛᅹࡊࠉ
ࡐࡿࡑࡿࢅ ncSCD1,ncSCD2 ࡛ࡊࡒࠊࡐࡊ࡙ࡐࡿࡼ㐿ఎᏄ࠿࢓࢜ࣂࣤ࢜ࣄ࡚࡜ࡡࡻ࠹࡝ᶭ⬗ࢅࡊ࡙࠷ࡾࡡ࠾ࢅ
ㄢ࡬ࡾࡒࡴ࡞ ncSCD1,ncSCD2 㐿ఎᏄࢅ◒ቪࡊ࡙ࡐࡡ⾪⌟ᆵࢅㄢ࡬ࡒࠊ షᠺࡊࡒ㐿ఎᏄ◒ቪᰬࡢⳞ⣊ࡡᙟ≟࡞␏ᖏࢅ♟ࡊࠉࡱࡒฦ⏍Ꮔࢅᙟᠺࡊ࡝࠷ࡆ࡛ࠉ㞜ᛮ୘⛩࡝࡜ࡡ⾪⌟ᆵࢅ♟ࡊࡒࠊࡱࡒࠉncSCD1ࠉncSCD2 ୦㐿ఎᏄ◒ቪᰬ㛣ࡡ⾪⌟ᆵࡢ㟸ᖏ࡞Ⰳࡂజࡒ≁ᚡࢅ♟ࡊࡒࠊࡆࡿࡼࡡࡆ
࡛ࡻࡽࠉncSCD1,ncSCD2 ࡢ࢓࢜ࣂࣤ࢜ࣄ࡞࠽࠷࡙ࡵᙟឺᙟᠺࡡㄢ⟿࡞㛭୙ࡊ࡙࠽ࡽࠉࡈࡼ࡞ ncSCD1,ncSCD2
ࡢྜྷୌ⤊㊪୕࡚ᶭ⬗ࡌࡾࡆ࡛࠿஢᝷ࡈࡿࡾࠊ

Characterization of signaling gene ncSCD1, ncSCD2 of Neurospora crassa
Masashi Kawamura, Akihiko Ichiishi(Toyo Univ, Fac. Life Sciences)P-20
adenylyl cyclase ✲↓ን␏ࢅᢒᅸࡌࡾ✲↓ን ␏మⱩἠᖶኯ 1),ἑහᙹ඼ 1),ᕝ⸠យᏄ 2),ᮟᒜ⫍Ꮔ 1) 1)㛭᮶Ꮥ㝌ኬ࣬ᕝ࣬∸㈻⏍࿤⛁Ꮥ⛁ 2)᪣࢝ࣚࢪ㸝ᰬ㸞Ⳟ⣊ᠺ㛏ࡡⴥࡊࡂ㐔࠷࢓࢜ࣂࣤ࢜ࣄᙟឺⓏ✲↓ን␏మ cr-1 ࡡᙟឺ␏ᖏࡡཋᅄ࠿ adenylyl cyclase 㐿ఎᏄࡡ✲↓ን␏࡞ࡻࡾࡵࡡ࡚࠵ࡾࡆ࡛࠿ሒ࿈ࡈࡿࡒ࠿ࠉcr-1 ࡡᠺ㛏࡛ᙟឺ␏ᖏࢅᅂᚗࡈࡎࡾᢒᅸ✲↓ን␏࠿㧏㢎ᗐ࡚㉫ࡆࡖࡒࠊࡐࡡ࠹ࡔࡡ 1 ᰬ hah ࡢ protein kinaseA ࡡ regulatory subunit ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ(MCB)ࡡ catalytic subunit࡛ࡡ⤎ྙ㒂న࡞㉫ࡆࡖࡒ✲↓ን␏మ࡚࠵ࡾࡆ࡛࠿ᮇ◂✪ᐄࡡᕝ⸠೒Ꮔࠉ♼ᓧㄌୌ࡞ࡻࡖ࡙➠ 3 ᅂࢤࣤࣆ࢒ࣝ
ࣤࢪ࡚ሒ࿈ࡈࡿࡒ࠿௑ᅂࡢ hah ࡛ࡢืࡡᢒᅸ✲↓ን␏㐿ఎᏄ wh ཀྵࡦ ah ࡞ࡗ࠷࡙ᙟឺࠉ㏻㙈⩄ࠉῺᗐវུᛮ
ࢅㄢ࡬ࡒࡡ࡚ࡆࡿࡼ࡞ࡗ࠷࡙ሒ࿈ࡌࡾࠊࡈࡼ࡞ ah ࡞ࡗ࠷࡙ࡢཋᅄ㐿ఎᏄ࠿ MCB ࡚࠵ࡽࠉhah ࡛ࡢ␏࡝ࡾ㹘
AMP ࡡ⤎ྙ㒂న࡞✲↓ን␏ࢅᣚࡗࡆ࡛࠿ฦ࠾ࡖࡒ࠿ࠉ
wh ✲↓ን␏మ࡞ࡗ࠷࡙ࡢཋᅄ㐿ఎᏄࢅ᥀⣬୯࡚࠵ࡾࠊSuppressor mutants which suppresse adenylyl cyclase mutation
Kouta Moizumi1), Masanori Kawauchi1),Aiko Kudo2),Tadako Murayama1)
1)Univ. of Kanto Gakuin   2)Asahi Glass co LTD

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㯔Ⳟ Aspergillus oryzae ࡢක࡞ᚺ➽ࡊฦ໩ࢅỬᏽࡌࡾ␂ᒜ⌦ᗀ㸡୯℀ᬓ஄, ᵵཾ⿩ḗ㑳, ᭯ᒱ Ꮥ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞࠘┘Ⓩ࠙㯔Ⳟ A. oryzae ࡢⓆ㓕⏐ᴏ୕᭩ࡵ᭯⏕࡝᚜⏍∸ࡡୌࡗ࡚࠵ࡽࠉࡐࡡ㓕⣪⏍⏐࣬ฦἢ⬗ࡡ㧏ࡈ࠾ࡼ␏⛸ࢰࣤࣂࢠ㈻⏍⏐ࡡฦ㔕࡞࠽࠷࡙ࡵ㔔さ࡝న⨠ࢅ༥ࡴ࡙࠷ࡾࠊࡈࡼ࡞ฦᏄ⏍∸ᏕⓏᡥἪࢅ⏕࠷ࡒ A. oryzae ࡡ㐿ఎᏄᶭ⬗ࡡよᯊ࡝࡜࠿ᗀࡂ⾔ࢂࡿ࡙࠷ࡾࠊࡊ࠾ࡊࡆࡿࡱ࡚ A. oryzae ࡞࠽࠷࡙ࠉ⎌ሾᅄᏄࡡୌࡗ࡚࠵ࡾක↯ᑏࡡ᭯↋࠿⏍⫩ࡷᙟឺฦ໩࡝࡜࡞ཀྵ࡯ࡌᙫ㡢࡞㛭ࡌࡾ▩ぜࡢ࡮࡛ࢆ࡜࡝࠷ࠊࡐࡆ࡚ᠻࠍࡢ㔕⏍ᰬ RIB40 ࢅ⏕࠷࡙ක↯ᑏࡡᙫ㡢ࢅㄢ࡬ࡒࠊ
࠘᪁Ἢ࡛⤎ᯕ࠙Potato Dextrose ࣈ࣭ࣝࢹᇰ㣬࡞࡙ⓉⰅක↯ᑏࡡᙫ㡢ࢅ⏍⫩ࡷฦ⏍Ꮔᙟᠺ࡞ࡗ࠷࡙ㄢ࡬ࡒ࡛ࡆࢀࠉ
㯔Ⳟ࡞࠽࠷࡙ิࡴ࡙ක↯ᑏୖ࡚ࡢ↋ᛮୠ௥ࡡ⬂Ꮔ࡚࠵ࡾฦ⏍Ꮔ࠿࡮࡛ࢆ࡜ᙟᠺࡈࡿ࡝࠷ࡆ࡛࠿ぜฝࡈࡿࡒࠊ
ྜྷࡋ⣊≟Ⳟ࡚࠵ࡾ A. nidulans ࡞࠽࠷࡙ࡢක↯ᑏୖ࡚ࡢ↋ᛮୠ௥ࠉ㟸↯ᑏୖ࡚ࡢ᭯ᛮୠ௥࠿ㄇᑙࡈࡿࡾࡆ࡛࠿
ฦ࠾ࡖ࡙࠷ࡾࠊࡐࡆ࡚ A. oryzae ࡛ A. nidulans ࢅẒ㍉ࡊࡒ࡛ࡆࢀ୕㏑ࡡオ㏑ࢅ⿤ࡘࡄࠉ஦ࡗࡡ⣊≟Ⳟ࡚㏣ࡡ⌟㇗࠿㉫ࡆࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡱࡒࠉ↯ᑏࡌࡾකࡡἴ㛏࡞ࡻࡾᙫ㡢ࢅㄢ࡬ࡾࡒࡴࠉ㉝Ⰵක࡛㟯Ⰵක࡞࠽ࡄࡾ⾪⌟ᆵࢅㄢ࡬ࡒࠊࡐࡡ⤎ᯕࠉ㉝Ⰵක࡞࠽࠷࡙ࡢࠉⓉⰅක࡛࡮࡯ྜྷୌ⛤ᗐࡡ⾪⌟ᆵࢅ♟ࡊࡒࡡ࡞ᑊࡊ࡙ࠉ㟯Ⰵක↯ᑏ࡞࠽࠷࡙ࡢᴗᗐࡡⓆⰾ㜴ᐐࠉ⏍⫩㜴ᐐ࠿☔ヾࡈࡿࡒࠊࡆࡿࡼࡡ⤎ᯕࡻࡽࠉකࡡᣚࡗᵕࠍ࡝ἴ㛏࡞ࡻࡾ␏࡝ࡖࡒࢨࢡࢻࣜ࠿㯔Ⳟࡡ⣵⬂හ࡞ఎࢂࡖ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
Light decides the differentiation in Aspergillus oryzae.
Rikou Hatakeyama, Tomoyuki Nakahama, Yujiro Higuchi, Manabu Arioka, and Katsuhiko Kitamoto (Dept. of
Biotechnol., Univ. of Tokyo㸞

P-22
THE smtA GENE ENCODING A SAM METHYL TRANSFERASE IS REQUIRED FOR
NORMAL DEVELOPMENT OF ASPERGILLUS NIDULANS
Kap-Hoon Han1, Hyo-Jung Kim, Jee Hyun Kim, and Dong-Min Han (Div. Biol. Sci. Wonkwang Univ. Iksan, 570-749,
Korea, 1Dept. Pharm. Engin. Woosuk Univ. Wanju, 565-701, Korea)
The smtA (SAM Methyl Transferase) gene encodes a methyl transferase carrying an S-adenosylmethionin binding
domain. The gene has 9 introns within an ORF consisted of 353 amino acids. The smtA gene in multi-copy can suppress
various mutations such as sndE80, silC188, silD6, silE181 and silF174, which are responsible for preferred
sexual development even in the conditions favored for asexual development. Both sexual and asexual sporulations werereduced and delayed by smtA deletion mutation, even under the conditions where either of those developments waspromoted. The mutant phenotypes suggest that the smtA gene is not essential for initiation of both sporulations.
However, it is necessary for development in normal rate. The smtA gene was expressed in high level during vegetative
growth and maintained upto the late stages of asexual and sexual development. The level of smtA mRNA was reducedin veA or fluG deletion mutant. The veA and the fluG deletion mutations were epistatic to smtA deletion mutation. These
results suggest that smtA acts downstream of veA or fluG which is known as a positive regulator of sexual or aesxual
development, respectively. And also they support the suggestion that SmtA functions in both sexual and asexual
development.

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Aspergillus oryzae ࡞࠽ࡄࡾ steA ࣓࣌ࣞࢡࡡᶭ⬗よᯊ᲻⏛ᐰெ, ➁හ㐠㞕㸝᮶ா㎨ᕝኬ㝌࣬⏍∸ᕝᏕ㸞
Aspergillus nidulans ࡡ steA ◒ቪᰬ࡚ࡢ᭯ᛮ⬂Ꮔࡡᙟᠺ࠿ᢒโࡈࡿࡾࡆ࡛࠾ࡼࠉsteA ࡢ᭯ᛮ⬂Ꮔࡡᙟᠺࢅโᚒࡌࡾ㌹෕ㄢ⟿ᅄᏄࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ࡛ࡊ࡙ሒ࿈ࡈࡿ࡙࠷ࡾࠊA. oryzae ࡡࢣࢿ࣑よᯊࡡ⤎ᯕࠉ᭯ᛮୠ௥ࢅᣚࡒ
࡝࠷ A. oryzae ࠾ࡼࡵ steA ࣓࣌ࣞࢡ࠿ぜ࠷ฝࡈࡿࡒࠊࡐࡆ࡚ࠉA. oryzae ࡞࠽ࡄࡾ steA ࣓࣌ࣞࢡࡡᶭ⬗ࢅㄢ࡬ࡾࡒࡴࠉsteA ࣓࣌ࣞࢡ㐛๨Ⓠ⌟ᰬ࡛ antisense mRNA Ⓠ⌟ᰬࢅష⿿ࡊࠉᐨኮᖲᯀᇰᆀࠉᾦమᇰᆀࠉᑚ㯇ࡨࡌࡱᇰᆀ࡚ࡡ⾪⌟ᆵࢅふᐳࡊࡒࠊࡐࡡ⤎ᯕࠉ࡜ࡡᇰᆀ࡞࠽࠷࡙ࡵ steA ࣓࣌ࣞࢡ㐛๨Ⓠ⌟ᰬࡢ WT ࡞Ẓ࡬࡙Ⳟ⣊ࡡ⏍⫩࠿㐔࠷ࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡱࡒࠉᐨኮᖲᯀᇰᆀ࡞࠽࠷࡙ࡢฦ⏍Ꮔࡡᙟᠺࡵᢒโࡈࡿ࡙࠷ࡒࠊࡆࡡ⤎ᯕࡢࠉA. nidulans ࡚ࡡሒ࿈࡛ୌ⮬ࡊ࡙࠽ࡽࠉ᭯ᛮୠ௥ࡡ࡝࠷ A. oryzae ࡚ࡵྜྷᵕࡡ⤎ᯕ࡛࡝ࡾࡆ࡛࠿᪺ࡼ࠾࡞
࡝ࡖࡒࠊࡈࡼ࡞ࠉᾦమࠉᑚ㯇ࡨࡌࡱᇰ㣬᫤࡞࠽ࡄࡾฦἢࢰࣤࣂࢠ㈻ࡡ SDS-PAGE ࣂࢰ࣭ࣤࢅㄢ࡬ࡒ࡛ࡆࢀࠉ
࡜ࡡᇰᆀ࡚ࡵ㐛๨Ⓠ⌟ᰬ࡞ࡢ≁␏Ⓩ࡝ࣁࣤࢺ࠿☔ヾ࡚ࡀࡒࠊ⌟ᅹࡆࡿࡼࡡࣁࣤࢺ࡞ࡗ࠷࡙ࠉ࣋ࣈࢲࢺ࣏ࢪࣆ
࢔࣭ࣤ࢝ࣈࣛࣤࢹἪ࡚よᯊ୯࡚࠵ࡾࠊ௧୕ࡡ⤎ᯕ࠾ࡼࠉA. oryzae ࡡ steA ࣓࣌ࣞࢡࡢⳞ⣊ࡡ⏍⫩࡞㛭୙ࡊࠉࡈࡼ࡞Ⳟమአࢰࣤࣂࢠ㈻ࡡⓆ⌟࡞ࡵ㛭୙ࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ

Functional analysis of steA homolog in Aspergillus oryzae
Hiroto Morita, Michio Takeuchi(Tokyo Univ. of Agriculture and Technology)

P-24
Sexuality and asexuality in Aspergillus species
Fabian A. Seymour1, Mathieu Paoletti1, Nanase Yamamoto2, Praveen R. Juvvadi2, Jun-ichi Maruyama2, Katsuhiko
Kitamoto2, David B. Archer1, Paul S. Dyer1  (1School of Biol., Univ. of Nottingham, UK, 2Dept. of Biotechnol., Univ.
of Tokyo㸞
The genus Aspergillus comprises species that reproduce by asexual and/or sexual means, and includes species withboth homothallic (sexually self fertile) and heterothallic (obligate outbreeding) breeding systems. Thus they provide
good model systems to study the evolution of sexual breeding systems and the molecular genetic basis of asexuality. In
addition, the aspergilli include species of importance in the biotechnology, food production and medical sectors. Thus
there are possible applied benefits to understanding the nature of reproduction in these species. We are investigating
reproduction in the supposed ‘asexual’ species A. fumigatus (an opportunistic pathogen) and A. oryzae (used in the
Asian food and biotechnology industries). A series of complementary genomic and experimental approaches have beenused to investigate possible reasons for asexuality in these species. Genome analysis of both A. fumigatus and A. oryzae
revealed the presence of a set of genes known to be involved with sexual reproduction in heterothallic filamentous
ascomycetes. Isolates used in the genome sequence projects contained either a MAT-2 high-mobility group gene or a
MAT-1 alpha-domain gene in A. fumigatus and A. oryzae respectively. Further experimental work identified thepresence of sexually compatible MAT-1 and MAT-2 isolates of A. fumigatus and A. oryzae in a screen of laboratory
isolates. Evidence for recombination was also gained in population genetic studies of A. fumigatus. Finally, expressionof certain key genes involved with sexual reproduction (mating-type, pheromone precursor and pheromone receptor
genes) was demonstrated for both species. Taken as a whole, these data suggest that both A. fumigatus and A. oryzae
have a recent evolutionary history of sexuality, and might retain the capacity to undergo sexual reproduction.

62

P-25
㯜㯔Ⳟ⏍⫩Ⳟమහࣈࣞࢷ࢛࣭࣑ࡡよᯊ♼ኯ㑳㸡ࢡ࢙ࣤ࣬ࢤࣤ࣬ࣀ㸡➁හ㐠㞕 㸝᮶ா㎨ᕝኬᏕ࣬ᚺ⏕⏍∸⛁Ꮥ⛁㸞๑ᅂࠉ㯜㯔Ⳟ A.oryzae RIB40 ࡡฦ⏍ᏄཀྵࡦⓆⰾฦ⏍ᏄⳞమහࡡࣈࣞࢷ࢛࣭࣑࡞ࡗ࠷࡙よᯊࡊࠉ⣑㸨㸥㸥಴ࡡࢪ࣎ࢴࢹࢅྜྷᏽࡊࠉ
ฦ⏍Ꮔ࡚ࡢ defense mechanism ࡞ᒌࡌࡾࢰࣤࣂࢠ㈻࠿ቌຊࡱࡒࡢ≁␏Ⓩ࡞Ꮛᅹࡊ࡙࠽ࡽࠉ
Ⓠⰾฦ⏍Ꮔ࡚ࡢฦ⏍Ꮔ࡞Ẓ࡬ metabolism 㛭㏻ࢰࣤࣂࢠ㈻࠿ቌຊࡊ࡙࠷ࡾࡆ࡛ࢅሒ࿈ࡊࡒ㸝ᖲᠺ㸦㸫ᖳࢤࣤࣆ࢒ࣝࣤࢪ㸞ࠊ㯜㯔Ⳟࡢ⏐ᴏ୕㔔さ࡝᚜⏍∸࡚࠵ࡽࠉ㏳ᖏ⏍⫩ᚃ᭿ࡡⳞమ࠿㯔࡛ࡊฺ࡙⏕ࡈࡿ࡙࠷ࡾࠊࡐࡆ࡚ࠉ
௑ᅂࠉᇰ㣬㛜ጙ㸦,㸧,㸨᪝┘ࡡ⏍⫩ᚃ᭿ࡡⳞమࢅ⏕࠷ࠉⳞమහࣈࣞࢷ࢛࣭࣑࡞ࡗ࠷࡙よᯊࢅ⾔ࡖࡒࠊᇰ㣬࡞ࡢSP ᾦమᇰᆀࢅ⏕࠷ࡒࠊᇰ㣬ᚃᅂ཭ࡊࡒⳞమࢅᾦమ✽⣪Ꮛᅹୖ࡚◒◃ࡊࠉᚋࡼࡿࡒࢰࣤࣂࢠ㈻࡞ࡗ࠷࡙஦ḗඔ㞹Ẵὃິࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ⏍⫩ᚃ᭿࡚ࡢࠉฦ⏍Ꮔ࣬Ⓠⰾฦ⏍Ꮔࡡ஦ḗඔ㞹Ẵὃິࣂࢰ࣭࡛ࣤࡢ᪺ࡼ࠾࡞
␏࡝ࡖ࡙࠷ࡒࠊ᪺ࡼ࠾࡞⏍⫩ᚃ᭿࡚ቌຊࡌࡾࢪ࣎ࢴࢹ㸦㸨಴ࠉ΅ᑛࡱࡒࡢᾐ⁓ࡌࡾࢪ࣎ࢴࢹ㸦㸨಴࡞ࡗ࠷࡙ࠉ
㯔Ⳟࢸ࣭ࢰ࣭࣊ࢪ࡛ MALDI-TOF-MASS ࢅ⏕࠷ࡒ࣋ࣈࢲࢺ࣏ࢪࣆ࢔࣭ࣤ࢝ࣈࣛࣤࢹἪ࡞ࡻࡽྜྷᏽࡊࡒࠊࡐࡡ⤎ᯕࠉ΅ᑛࡱࡒࡢᾐ⁓ࡌࡾࢪ࣎ࢴࢹ࡛ࡊ࡙ Heat shock proteinࠉᶭ⬗ᮅ▩ࢰࣤࣂࢠ࡝࡜࠿ヾࡴࡼࡿࡒࠊୌ᪁ࠉ
ᇰ㣬㛜ጙ㸨᪝┘ࡡ⏍⫩ᚃ᭿ࡡⳞమ࡚ࡢ௥ㅨ㛭㏻ࡡࢰࣤࣂࢠ㈻࠿ቌຊࡊࠉྜྷୌࡡࢰࣤࣂࢠ㈻࠿々ᩐࡡࢪ࣎ࢴࢹ
࡛ࡊ࡙᳠ฝࡈࡿࡾࡆ࡛࠾ࡼಞ㣥ࢅུࡄ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ⏍⫩Ⳟమ࡚ቌຊࡊࡒⳞమහࢰࣤࣂࢠ㈻ࡢࠉ
ሲวࡽ㯔ࡡ㓕⣪࡛ࡊ࡙㔔さ࡝ᙲ๪ࢅᯕࡒࡊ࡙࠷ࡾࡵࡡ࡛⩻࠻ࡼࡿࡒࠊ
Proteome analysis of intracellular proteins from A.oryzae RIB40.
Taro Ko, Nguyen Cong Ha, Michio Takeuchi(Tokyo Univ. of Aguriculture and Technology)

P-26⣊≟Ⳟ࡞࠽ࡄࡾ APase 㐿ఎᏄࡡኣᵕᛮ
ᒱᮇ⥜Ꮔ㸡➁හ㐠㞕 㸝᮶ா㎨ᕝኬᏕ࣬㎨࣬ᚺ⏍⛁㸞
㯔Ⳟ㸝Aspergillus oryzae RIB40㸞ࡡࢣࢿ࣑よᯊ࡞ࡻࡖ࡙ࠉࢣࢿ࣑හ࡞ࡢሲᇱ㒼า࠾ࡼ᥆ᏽࡈࡿࡾ࢓࣐ࢿ㓗㒼าࡻࡽ࢓ࢪࣂࣜࢷ࢕ࢴࢠࣈࣞࢷ࢓࣭ࢭ㸝APase㸞࡛⩻࠻ࡼࡿࡾ㐿ఎᏄ࠿ኣᩐᏋᅹࡌࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡆ
ࡿࡼࡡ㐿ఎᏄ࠿ A. oryzae RIB40 ᰬ௧አࡡ A. oryzae ᰬ࡞Ꮛᅹࡌࡾࡡ࠾ࢅㄢ࡬ࡾࡒࡴࠉA. oryzae RIB40 ࡡ APase㐿ఎᏄධ㛏ࢅࣈ࣭ࣞࣇ࡛ࡊ࡙ A. oryzae ᰬ㸩⛸㢦࡛ A. sojae ࡞ࡗ࠷࡙ࢦࢧࣤよᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ௙ࡡ A.
oryzae ᰬࡷ A. sojae ࡞ࡵ A. oryzae RIB40 ࡡᣚࡗ APase 㐿ఎᏄࡢᏋᅹࡊ࡙࠷ࡒࠊA. oryzae ᰬࡡ APase 㐿ఎᏄࡢA. oryzae RIB40 ࡡ APase 㐿ఎᏄ࡛ྜྷࡋన⨠࡞ࣁࣤࢺ࠿ฝ⌟ࡊࡒࠊࡊ࠾ࡊࠉA. sojae ࡞Ꮛᅹࡌࡾ APase 㐿ఎᏄࡡ࠹ࡔ㸬⛸㢦ࡢ A. oryzae RIB40 ࡡ APase 㐿ఎᏄ࡛ࡢ␏࡝ࡾన⨠࡞ࣁࣤࢺ࠿ฝ⌟ࡊࡒࠊࡈࡼ࡞ࠉA. oryzae RIB40
ࡡ APase 㐿ఎᏄࡡ࠹ࡔ㸦⛸㢦ࡢ A. sojae ࡚ࣁࣤࢺ࠿ฝ⌟ࡊ࡝࠾ࡖࡒࠊࡱࡒࠉ㒼า࠿ࡌ࡚࡞ሒ࿈ࡈࡿ࡙࠷ࡾ A.
nidulans ࡞ࡗ࠷࡙ࡵྜྷᵕ࡞ࢦࢧࣤよᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉA. oryzae ࡞ࡢᏋᅹࡌࡾ࠿ࠉA. nidulans ࡞ࡢᏋᅹࡊ࡝࠷ APase 㐿ఎᏄࡡᏋᅹ࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡈࡼ࡞ࠉࢣࢿ࣑㒼า࠿᪺ࡼ࠾࡞࡝ࡖࡒ⣊≟Ⳟࡡ APase 㐿ఎᏄ࡞ࡗ࠷࡙ࡵ A. oryzae ࡞࠽ࡄࡾ APase 㐿ఎᏄࡡሲᇱ㒼า࠾ࡼ᥆ᏽࡈࡿࡾ࢓࣐ࢿ㓗㒼า᝗ሒࢅࡵ࡛࡞よᯊࢅ⾔࠷ APase ࡡ⣌⤣ᶖࢅషᠺࡊࡒࠊࡐࡡ⤎ᯕࠉ⣊≟Ⳟࡡ APase ࡢኬࡀࡂ㸩ࡗࡡࢡ࣭ࣜࣈ࡞ฦ㢦࡚ࡀࡾࡆ࡛࠿♟ြࡈࡿࡒࠊDiversity of APase genes from Filamentous fungi.
Ayako okamoto, Michio Takeuchi(Tokyo Univ.of Agriculture and Technology)

63

P-27
糸状菌 Aspergillus nidulans のプロセッシング酵素遺伝子 kexB 破壊株の解析
ୌᰏಆ௒ 1, ཿᕖ೸ኯ㑳 1, Ềㆺ἖ 2, ⸠ᒱᬓ์ 1, ᚠᒱ᪸ᩝ 2, ஫࿝ົஒ 2, 㜷㒂ᩏᝃ 1 (᮶໪ኬ㝌㎨࣬1 ᚺ⏍⛁, 2 ⏍∸⏐ᴏ๭⏍)
Ở ᠻࠍࡢࠉ⣊≟Ⳟ⏍మහ࡚ࡡฦἢࢰࣤࣂࢠ㈻ࡡࣈࣞࢬࢴࢨࣤࢡᶭᵋࡡよ᪺ࢅ┘ᣞࡊࠉࡆࡿࡱ࡚࡞⣊≟Ⳟ
Aspergillus nidulans ࡡࢪࣇࢲࣛࢨࣤᵕࣈࣞࢬࢴࢨࣤࢡ㓕⣪㐿ఎᏄ kexB ◒ቪᰬ (ΔkexB ᰬ) ࢅ⏕࠷ࡒࢹࣚࣤ
ࢪࢠࣛࣈࢹ࣭࣑よᯊࢅ⾔ࡖ࡙ࡀࡒࠊA. nidulans ΔkexB ᰬࡢࠉCell Intgrity ⤊㊪ (PKC ⤊㊪) ࡞␏ᖏࢅࡵࡒࡼࡊࠉ
㔕⏍ᰬ࡛Ẓ࡬࡙㟸ᖏ࡞ࢤࣤࣂࢠࢹ࡝ࢤࣞࢼ࣭ࡊ࠾ᙟᠺࡊ࡝࠷࠿ࠉ㧏ᾈ㏩ᅸ᮪௲ୖ࡚ᇰ㣬ࡌࡾ࡛⾪⌟ᆵ࠿ᚗᖉࡊࡒࠊࡱࡒࠉA. oryzae ࡡࢡࣜࢤ࢓࣐࣭ࣚࢭ GlaA ࡡࣃࣤࢩ㡷ᇡ࡞ KexB ࡡヾㆉࢦ࢕ࢹ Lys-Arg ࢅᑙථࡊࠉ
␏⛸ࢰࣤࣂࢠ㈻࡛ࡊ࡙ࢱࢼ࢓ࣝࣜࢣࣤ Derf7 ࢅ㏻⤎ࡈࡎࡒ⼝ྙࢰࣤࣂࢠ㈻ࡢࠉA. nidulans 㔕⏍ᰬࡡᇰ㣬୕ΰ
୯࡞ࡢ N ᮆ❻ഁり፳ࢺ࣒࢕࡛ࣤ C ᮆ❻ഁ␏⛸ࢰࣤࣂࢠ㈻࠿ว᩷ࡈࡿࡒ≟ឺ࡚ฦἢࡈࡿࡒ࠿ࠉΔkexB ᰬ࡚ࡢࡐࡡࡱࡱࡡᙟ࡚ฦἢࡈࡿࡒࠊ⮾࿝῕࠷ࡆ࡛࡞ࠉΔkexB ᰬࢅ㧏ᾈ㏩ᅸ᮪௲ୖ࡚ᇰ㣬ࡌࡾ࡛ࣈࣞࢬࢴࢨࣤࢡ⬗ງ࠿ᚗᖉࡊࡒࠊLys-Arg ࢅ Ala-Ala ࡞⨠ᥦࡊࡒ࡛ࡆࢀࠉ㏳ᖏᇰᆀ࡚ࡢࠉ㔕⏍ᰬ࣬ΔkexB ᰬභ࡞ࣈࣞࢬࢴࢨࣤࢡ
ࢅධࡂུࡄ࡝࠾ࡖࡒࠊ㧏ᾈ㏩ᅸᇰᆀ࡚ࡢഷ࠾࡞ࣈࣞࢬࢴࢨࣤࢡ࠿ぜࡼࡿࡒ࠿ࠉLys-Arg మࡻࡽࡵᑛ࡝࠾ࡖࡒࠊ
ࡆࡿࡼࡡࡆ࡛ࡢࠉ㧏ᾈ㏩ᅸ᮪௲ୖ࡚ࡡࡲ഼ࡂ KexB ᵕࡡᇱ㈻≁␏ᛮࢅᣚࡗࣈࣞࢬࢴࢨࣤࢡ㓕⣪࠿Ꮛᅹࡌࡾࡆ
࡛ࢅ♟ြࡊ࡙࠷ࡾࠊ⌟ᅹࠉᠻࠍࡢ⛸ࠍࡡ᪁Ἢࢅ⏕࠷ࠉࡆࡡࣈࣞࢬࢴࢨࣤࢡ㓕⣪ࡡྜྷᏽࢅモࡲ࡙࠷ࡾ࡛ࡆࢀ࡚
࠵ࡾࠊCharacterization of the filamentous fungus Aspergillus nidulans kexB disruptant.
Shunsuke Ichiyanagi, Kentaro Furukawa, Osamu Mizutani, Tomonori Fujioka, Masafumi Tokuoka, Katsuya Gomi and
Keietsu Abe (Tohoku Univ., Grad. Sch. Agri. Sci.)

P-28⣊≟Ⳟ Aspergillus nidulans ࡞࠽ࡄࡾ class III ࢞ࢲࣤฦよ㓕⣪ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄࡡᶭ⬗よᯊᒜᓧᬍ୓㸡ᇷහ⿩஄ࠉኯ⏛᪺ᚠ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞⣊≟Ⳟ Aspergillus nidulans ࡡ⣵⬂ባࡡ୹さᵋᠺᠺฦࡡୌࡗࡢ࢞ࢲ࡚ࣤ࠵ࡽࠉࡐࡡྙᠺ࣬ฦよࡢࠉA. nidulansࡡᙟឺᙟᠺ࡞࠽࠷࡙ཚᐠ࡞โᚒࡈࡿ࡙࠷ࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊA. nidulans ࡡࢣࢿ࣑࡞ࡢࠉ⢶㈻ฦよ㓕⣪ฦ㢦ࣆ࢒࣐࣭ࣛ18 ࡞ᒌࡌࡾ࢞ࢲࣤฦよ㓕⣪ࢅࢤ࣭ࢺࡌࡾ࡛⩻࠻ࡼࡿࡾ㒼า࠿ 18 ಴Ꮛᅹࡊࠉࡐࡡ࠹ࡔ 3 ಴࠿ class IIIࠉ
15 ಴࠿ class V ࡞ᒌࡌࡾ࢞ࢲࣤฦよ㓕⣪㐿ఎᏄࢅࢤ࣭ࢺࡊ࡙࠷ࡾ࡛᥆ᏽࡈࡿࡾࠊ௑ᅂࠉclass III ࢞ࢲࣤฦよ㓕⣪ࢅࢤ࣭ࢺࡌࡾ࡛⩻࠻ࡼࡿࡾ㐿ఎᏄ chiAࠉchiHࠉchiR ࡞ࡗ࠷࡙よᯊࢅ⾔ࡖࡒࠊchiAࠉchiHࠉchiR ࡡ྘༟≺◒ቪᰬࠉࡈࡼ࡞ chiAࠉchiH ࡱࡒࡢ chiAࠉchiR ࡡ஦㔔◒ቪᰬ࡚ࡢࠉ㔕⏍ᆵᰬ࡛ྜྷᵕࡡ⏍⫩ࢅ♟ࡊࡒࠊࡊ࠾ࡊ chiHࠉ
chiR ࡡ஦㔔◒ቪᰬࡢཱིᚋ࡚ࡀࡍࠉࡆࡿࡼ୦㐿ఎᏄࡡḖ኶ࡢ⮬Ṓ࡚࠵ࡾྊ⬗ᛮ࠿⩻࠻ࡼࡿࡒࠊࡐࡆ࡚ chiR ࡡ◒ቪᰬ࡞࠽࠷࡙ࠉchiH ࡡⓆ⌟ࢅ A. nidulans ࡞࠽࠷࡙ᇰᆀࡡ⅛⣪″࡞ࡻࡽⓆ⌟ࡡโᚒྊ⬗࡝ alcA ࣈ࣓࣭ࣞࢰ࣭ࡡ
ᨥ㒼ୖ࡚ᢒโࡊࡒ࡛ࡆࢀࠉ㔕⏍ᆵᰬ࡞Ẓ࡬Ⳟ⣊ࡡ⏍⫩࡞㐔ࡿ࠿ぜࡼࡿࡒࠊࡆࡡᰬ࡞࠽࠷࡙ࡈࡼ࡞ chiA ࢅ◒ቪࡊ࡙ࡵᙫ㡢ࡢ࡝࠾ࡖࡒࠊࡆࡿࡼࡡࡆ࡛࠾ࡼ chiH ࡛ chiR ࡢࠉchiA ࡛ࡢ≺❟ࡊࡒப࠷࡞㔔々ࡊࡒᶭ⬗ࢅ᭯ࡊ࡙࠽ࡽࠉ⏍⫩࡞㔔さ࡝ᙲ๪ࢅᯕࡒࡊ࡙࠷ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ

Functional analysis of class III chitinase genes in Aspergillus nidulans
Harutake Yamazaki, Hiroyuki Horiuchi and Akinori Ohta
(Dept. of Biotechnol., Univ. of Tokyo)

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Aspergillus tamarii ⏜ᮮࡡ niaD 㐿ఎᏄࡡᵋ㏸
ᮄᮟኣỜ 1 ᴃᮇ៿ୌ 1 ໪ᮇ์⾔ 2  㕝ᮄ⪵ 1 ᯵ᮄ㇇ 1 㸝1 ≺Ἢ࣬㣏⥪◂, 2 យ▩⏐ᢇ◂࣬㣏ဗᕝᢇ㸞
࠘┘Ⓩ࠙㓲ἔ⢉ࡢධᅗ࡚ᖳ㛣⣑ 10 ୒ࢹࣤ࠿㓲ἔ⿿㏸ࡡ๧⏐∸࡛ࡊ࡙ᤴฝࡈࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉ㓲ἔ⢉ࡢሲฦࢅኣ㔖࡞ྱࡳࡒࡴࠉࡐࡡ᭯ຝ࡝ฌ⌦᪁Ἢ࠿☔❟ࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚ࠉ㧏ฦᏄኣ⢶㢦ฦよ㓕⣪ࢅ⏍⏐ࡌࡾ⣊≟Ⳟࢅ⏕࠷࡙ࠉ㓲ἔ⢉ࢅฦよฌ⌦ࡌࡾ஥ࢅ⩻࠻ࡒࠊ᭩ิ࡞ᩐᰬࡡ㯔Ⳟࡻࡽ㓲ἔ⢉୕࡚⏍⫩࡚ࡀࡾ A. tamarii
ST-2 ᰬࢅ㐽ᢜࡊࡒࠊḗ࡞ᙟ㈻㌹ᥦ࡞ࡻࡖ࡙ኣ⢶㢦ฦよ㓕⣪㐿ఎᏄࢅᑙථࡌࡾࡒࡴ࡞ࠉᮇᰬࡡ niaD -ᰬࢅㄇᑙࡊࡒࠊᚋࡼࡿࡒ A. tamarii ST-2 niaD -ᰬ࡞ A. oryzae ࡡ niaD 㐿ఎᏄࢅ㏻⤎ࡊࡒࣈࣚࢪ࣐ࢺࢅᑙථࡊࡒ࠿ࠉᙟ㈻㌹ᥦమࢅᚋࡾࡆ࡛ࡢฝᮮ࡝࠾ࡖࡒࠊ ࡐࡡࡒࡴࠉᮇ◂✪࡚ࡢᙟ㈻㌹ᥦ࣏࣭࡛࢜ࡊ࡙ A. tamarii ST-2 ᰬ⏜ᮮࡡ niaD 㐿ఎᏄࢅฦ㞫ࡊࠉࡐࡡᵋ㏸ࡡよ᪺ࢅ⾔ࡖࡒࠊ
࠘᪁Ἢཀྵࡦ⤎ᯕ࠙A. oryzae ཀྵࡦ㎾⦍⛸ࡡ niaD 㐿ఎᏄࡡሲᇱ㒼า࡞ᇱࡘ࠷࡙スゝࡊࡒ PCR ࣈࣚ࢕࣏࣭ࢅ⏕࠷
࡙ࠉA. tamarii ST-2 ࢣࢿ࣑ DNA ࢅ㗢ᆵ࡞ࡊ࡙ degenerated PCR ࢅ⾔ࡖࡒࠊniaD ORF ࡡ N ᮆ❻ཀྵࡦࠉC ᮆ❻㡷ᇡ࡞┞ᙔࡌࡾࡐࡿࡑࡿ⣑ 250bp ࡡ DNA ᩷∞ࢅཱིᚋࡊࡒࠊࡆࡿࡼࡡሲᇱ㒼าࡢ A. oryzae ࡡ niaD 㐿ఎᏄ࡛ 90%௧୕ࡡ㧏࠷┞ྜྷᛮࢅ♟ࡊࡒࠊࡐࡆ࡚ࠉࡆࡿࡼࡡ A. tamarii ⏜ᮮࡡ DNA ࣆࣚࢡ࣒ࣤࢹ࡛ A. oryzae niaD 㐿ఎᏄࡡሲᇱ㒼าࢅᇱ࡞ࣈࣚ࢕࣏࣭ࢅスゝࡊࠉPCR ࡞ࡻࡖ࡙ቌᖕࡊࡒ࡛ࡆࢀࠉA. tamarii niaD ࡡ⩳ゼ㡷ᇡࡡධ㡷ᇡ⣑ 3kbࠉ୕Ὦᇡ⣑ 0.4kb ཀྵࡦࠉୖὮᇡ⣑ 0.9kb ࡞┞ᙔࡌࡾ㡷ᇡࢅཱིᚋ࡚ࡀࡒࠊ
 A. tamarii ࡛ A. oryzae ࡡ⩳ゼ㡷ᇡࡡ┞ྜྷᛮࡢࠉሲᇱ㒼า࡚⣑ 91%ࠉ࢓࣐ࢿ㓗㒼า࡚⣑ 98%࡚࠵ࡖࡒࠊ
A. tamarii niaD ࡡ 3’㟸⩳ゼ㡷ᇡ࡞඼ᆵⓏ࡝࣎ࣛ A ௛ຊ㒼า(AATAAA)ࡢヾࡴࡼࡿ࡝࠾ࡖࡒ࠿ࠉ㢦జࡊࡒ㒼า(AATATA,AATACA)࠿ 2 ࢜ᡜヾࡴࡼࡿࡒࠊ࠷ࡂࡗ࠾ࡡ㓕ẍࡷ࢜ࣄ࡚ࢰ࣭࣐ࢾ࣭ࢨࣘࣤࢨࢡࢻ࡛ࣜࡊ࡙ሒ࿈ࡈ
ࡿ࡙࠷ࡾ㒼า(TACATTA, CATGTCTT)ࡢ 1 ࢜ᡜ(CATGTCTT ࡡࡲ)ヾࡴࡼࡿࡒࠊ
Structural analysis of niaD from Aspergillus tamarii
Tae Kimura1, Ken-Ichi Kusumoto1, Noriyuki Kitamoto2, Satoshi Suzuki1, Yutaka Kashiwagi1(1Natl. Food Res. Inst., 2Food Res. Center, Aichi Ind. Technol. Inst.)

P-30
Insights into RIP and DNA methylation in the Aspergillus section Flavi complex

Heather A.

Lee , Maria Dolores Montiel and David B. Archer  㸝 Institute of Genetics, School of Biology, University of
Nottingham, University Park, Nottingham, NG7 2RD 㸞
DNA methylation of cytosine is an epigenetic mechanism found in many eukaryotic genomes, with the dual roles ofgene regulation and protection of the genome. Control and function of DNA methylation are still largely unknown infungi and, apart from detailed work in Ascobolus immersus and Neurospora crassa, few studies have been conducted inother species. In N. crassa DNA methylation is involved in a process called Repeat-Induced Point Mutation (RIP),
which acts at a precise stage in the sexual cycle, detecting sequence duplications and introducing C:G and T:Atransitions leading to dense cytosine methylation. Recently RIP-like transitions have been reported in transposons inAspergillus nidulans and A. fumigatus, but Aspergillus species were thought to be devoid of DNA methylation, although
this has now been contradicted in work detecting methylation by HPLC for A. flavus. We isolated a transposase
sequence from A. parasiticus using a DNA methyl-binding column. The sequence has 67% identity with Tan 1 from A.
niger and is present in at least 20 copies in the A. oryzae genome sequence database. Comparison of these copies
indicates the presence of RIP-like transitions. The pattern was similar to RIP changes in N. crassa but was less dense.
Although the original sequence was isolated from a methyl-binding column, no evidence of methylation was found bySouthern Blotting. The preference for CpA to TpA changes could increase the introduction of termination codons (TAA
and TAG) leading to gene inactivation by mutation alone. It is also possible that this light form of RIP is a source ofgenetic diversty.

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㯔Ⳟࡡ G ࢰࣤࣂࢠ㈻භᙲᆵ cAMP ུᐖమ┞ྜྷ㐿ఎᏄ㕝ᮄ ⪵㸡➁ㆺ༡Ꮔ, ᮄᮟኣỜ, ᮿୖ┷⏜⨶, ᴃᮇ៿ୌ, ᯵ᮄ ㇇ 㸝㣏⥪◂㸞⣵⬂ࡢᵕࠍ࡝⣵⬂አ࠾ࡼࡡಘྒࢅུᐖࡊ⣵⬂හ࡫ࡡಘྒఎ㐡ࢅ⾔࡝࠷⎌ሾ࡫ࡡ㐲ᚺࢅᅒࡖ࡙࠷ࡾࠊ㯔Ⳟ㎾⦍
⛸࡚ࡢࠉAspergillus nidulans ࡡ◂✪࡞ࡻࡽ G ࢰࣤࣂࢠ㈻භᙲᆵུᐖమ㸝G protein-coupled receptor: GPCR㸞ࡡᏋ
ᅹ࠿᪺ࡼ࠾࡞࡝ࡖࡒ࠿ࠉ⣊≟Ⳟ࡞࠽ࡄࡾ GPCR ཀྵࡦࡐࡡࣛ࢝ࣤࢺ࡞㛭ࡌࡾ▩ぜࡢᑛ࡝࠷ࠊᠻࠍࡢࠉ㯔Ⳟࢣࢿ
࣑୯࡞ 6 ⛸ࡡ GPCR ᵕ㒼าࢅぜฝࡊࠉࡐࡡ࠹ࡔ cAMP ུᐖమ┞ྜྷ㒼า࠿Ⓠ⌟ࡊ࡙࠷ࡾ஥ࢅ☔ヾࡊࡒࠊࡱࡒࠉ
A. oryzae RIB40 ᰬࡡࢣࢿ࣑ DNA ࢅ㗢ᆵ࡛ࡊࡒ PCR ࡞ࡻࡽᮇ㐿ఎᏄࢅྱࡳ DNA ᩷∞ࢅཱིᚋࡊࡒ㸝2005 ᖳᗐ㎨ⰹ໩Ꮥఌኬఌ㸞ࠊ௑ᅂࡢࠉᮇ㐿ఎᏄࡡ 5̓࠽ࡻࡦ 3̓ᮆ❻㒼าࢅ☔ヾࡊࠉ㟸⩳ゼ㡷ᇡ㒼าࡵྱࡴࡒධ㛏 cDNAࡡཱིᚋ࡛ࡐࡡධሲᇱ㒼าࢅỬᏽࡊࡒࡡ࡚ሒ࿈ࡌࡾࠊ

G protein-coupled cAMP receptor homologue of Aspergillus oryzae
Satoshi Suzuki, Hiroko Taketani, Tae Kimura, Mayumi Matsushita Ken-Ichi Kusumoto, Yutaka Kashiwagi(Natl. Food Res. Inst.)

P-32
Dikaryon ࡡฦ⿛࡞࠽ࡄࡾࢠࣚࣤࣈࡡណ⩇ᒱᓧᏏ᫆㸡୸⩒ಞ㌗ 㸝࠾ࡍࡈ DNA ◂✪ᡜ࣬᯹Ⰵమᶭ⬗㡷ᇡ➠ୌ㸞࢞ࢿࢤࡢฦ⿛ࡇ࡛࡞ dikaryon ࡡ㸧ࡗࡡᰶࢅࡐࡿࡑࡿฦ㒼ࡈࡎࡾࡒࡴ࡞ࢠࣚࣤࣈ࡛࿣ࡣࡿࡾᰶ࠿㏳ࡾࣁ࢕ࣂ
ࢪࢅᙟᠺࡌࡾࠊᠻࠍࡢฦ⿛㓕ẍࢅ⏕࠷࡙ dikaryon ࡡฦ⿛ᶭᵋࢅよᯊࡊ࡙࠷ࡾ࠿ࠉ໗ㄵࢠࣚࣤࣈࡢ࡝࠷ࠊࢠࣚ
ࣤࣈࡡ࡝࠷࢞ࢿࢤࡵ࠵ࡽࠉࡐࡡᰶฦ⿛ࡡᵕᘟࡢฦ⿛㓕ẍࡡ࡛ࡻࡂజ࡙࠷ࡒࠊฦ⿛㓕ẍ dikaryon ࡡฦ⿛࡚ࡢࠉ
᭩ิ A,A,B,B ࡛୩ࢆࡓፃᰶ࠿ A,B,A,B ࡡ㡨࡞ථࡿ᭨ࢂࡾࡡࢅ anaphase B ୯⛤ࡱ࡚㜴ࡳࡵࡡ࠿࠵ࡾࠊࡐࡿ࠿ப
࠷ࡡᰶࡐࡡࡵࡡࡓ࡛ࡌࡾ࡛ࠉฦ⿛㓕ẍࡢ࡜࠹ࡷࡖ࡙ࢠࣚࣤࣈࡵ౐ࢂࡍ࡞ፃᰶྜྷኃ࠿ப࠷ࡡ㝸ባ࡛࡝ࡾࡡࢅᅂ
㑂ࡌࡾࡡ࠾ࠊࡐ࠹⩻࠻࡙よᯊࡊ࡙ࡀࡒ࠿ࠉណአ࡞ࡵፃᰶࡡථࡿ᭨ࢂࡽࢅ㜴ࡳࡵࡡࡢࠉᐁࡢ⣵⬂㈻ฦ⿛࢓ࢠࢹ
࣐࢛ࢨࣤ཭⦨⎌ࡡᠺ⇅ࢲ࢘ࢴࢠᶭ⬗࡚࠵ࡖࡒࠊ⌟᫤Ⅴ࡚ࡢࠉᰶ࡜࠹ࡊࡡ∸⌦Ⓩ㝸ባࢅ㑂ࡄࡾᶭᵋ࠿࡝࠷࡛ࡢ
ゕ࠻࡝࠷࠿ࠉ∸⌦Ⓩ㝸ባ࡞࡝ࡖ࡙࠷ࡾ࡛࠷࠹ッᣈࡵ࡝ࡂ࡝ࡖࡒࠊࢠࣚࣤࣈࡡᙲ๪ࡢப࠷ࡡᰶ࠿∸⌦Ⓩ㝸ባ࡛࡝ࡾࡡࢅᅂ㑂ࡌࡾࡆ࡛ࡓ࡛࠷࠹ぜ᪁࠿ࡈࡿ࡙ࡀࡒ࠿ࠉᮇᙔ࡞ࡐ࠹࡝ࡡ࠾ࠊࢠࣚࣤࣈ࠿ᏄᄙⳞ㢦ࡡ crozier ᵋ㏸࡞㢦జࡊ࡙࠷ࡾࡆ࡛ࡢ௧๑࠾ࡼᣞᦤࡈࡿ࡙ࡀࡒࠊࡆࡡᵋ㏸ࡢ dikaryon ࠿ᰶ⼝ྙࡌࡾ├๑࡞㸦ࡗࡡ dikaryon ࡛㸧ࡗࡡ monokaryon ࢅ⏍ࡍࡾࡵࡡ࡚࠵ࡾ࠿ࠉፃᰶ࡜࠹ࡊࡡ∸⌦Ⓩ㝸ባࢅᅂ㑂ࡌࡾណ࿝࠿࠵ࡾ࡛ࡢᛦ࠻࡝࠷ࠊ㸧ࡗࡡ monokaryon ࡢ්ࡦ᥃ྙࡊ࡙ dikaryon ࡛࡝ࡽࠉḗࡡ crozier ࢅᙟᠺࡌࡾࠊࡗࡱࡽ crozier ࡢ᥃ྙ࡞ࡻࡖ࡙㸧ࡗࡡᰶࡡ᥃ྙᆵ࠿ࣉࢷ࡚ࣞ࠵ࡾࡆ࡛ࢅ☔ヾࡌࡾᶭ⬗ࢅᯕࡒࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾࠊࢠࣚࣤࣈࡵྜྷᵕ࡞ࠉฦ⿛
ࡡࡒࡦ࡞᥃ྙ࡞ࡻࡖ࡙ᰶ࠿ࣉࢷ࡚ࣞ࠵ࡾࡆ࡛ࢅ☔ヾࡌࡾࡒࡴ㸝ᐁ㝷ࢠࣚࣤࣈᙟᠺࡢ᥃ྙᆵ㐿ఎᏄ࠿ࣉࢷ࡚ࣞ
࠵ࡾࡆ࡛࡞౪Ꮛࡊ࡙࠷ࡾ㸞ࡡ⿞⨠࡚ࡢ࡝࠾ࢀ࠹࠾ࠊ

Role of clamp connection for conjugate division of dikaryon
Koei Okazaki, Osami Niwa
(Kazusa DNA Research Institute)

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smuH501, a second mutation found in the original uvsH77 mutant strain that involved in DNA
replication checkpoint control in Aspergillus nidulans
Mee-Jeong Cha, Sun-Hee Noh, Nak-Jung Kwon, and Suhn-Kee Chae (Dept. Biochem. and Biomed RRC, Paichai
Univ. Daejeon, 302-735 Korea)
smuH (suppressor of mutagen-sensitivity of uvsH77) was originally isolated as a DNA fragment complementing
partially the MMS-sensitivity of uvsH77 mutant during the course of the uvsH gene (a yeast RAD18 homolog) cloning
in A. nidulans. ǼsmuH as well as the original uvsH77 mutant strain exhibited high sensitivities to camptothecin (CPT),a topoisomerase I inhibitor. However, during the analysis of meiotic progenies in uvsH77 heterozygotic crosses, asecond mutation causing high CPT-sensitivity was identified. The second mutation, called smuH501 was shown to be amutant allele of smuH. The mutation site was also identified in smuH501, changing the 56th lysine residue to a stopcodon. Therefore, smuH was redefined as "Second Mutation in uvsH77". smuH501 alone showed high CPT-

but veryslight MMS-sensitivities, while uvsH77 without smuH501 exhibited no CPT- and much reduced MMS-sensitivities thanthe original uvsH77 mutant. Null mutants of uvsH77 showed MMS-sensitivity similar to those shown in uvsH77(without smuH501), uvsH304, and uvsH311 mutants. Synergistic interaction between smuH and uvsH was evident. In
addition, smuH501 showed synergistic interaction with uvsF201 encoding replication factor C. SmuHp exhibited theamino acid sequence similarity to Tof1 of S. cerevisiae and Swi1 of S. pombe, which are involve in DNA replication
checkpoint control.

P-34⺧කࢰࣤࣂࢠᵾㆉࢅ⏕࠷ࡒ Aspergillus nidulans ࢰࣤࣂࢠ㈻ࡡ⣵⬂࿔᭿ንິࡡࣚ࢕ࣇ
࢕࣒࣭ࢩࣤࢡ㸯 II ᆵ topoisomerase ࢅ୯ᚨ࡞1⣒ ᭰ᾀ㸡ᕖᓃ⨶౅㸡ⳝờ㡝ᙢ㸡2Berl R. Oakley㸡3 ᇷᑹူஒ㸝1 ྞཿᒁኬ㝌༈⣌࣬ฦᏄᵾⓏ㸡2 ࢛ࣀ࢕࢛ᕗ❟ኬ࣬ฦᏄ㐿ఎ㸡3 ᚠᓞኬ㝌࣬ࣉࣜࢪࣁ࢕࢛㸞ϩᆵ topoisomerase㸝topoII㸞ࡢ DNA ࡡ㧏ḗᵋ㏸ࢅㄢ⟿࣬โᚒࡌࡾ㓕⣪࡚࠵ࡾࠊࡆࡡ㓕⣪Ὡᛮࢅ㜴ᐐࡌࡾⷾ๠ࡢᢘ࠿ࢆ๠࡝࡜࡞ฺ⏕ࡈࡿ࡙࠷ࡾ࠿ࠉᐁ㝷࡞⣵⬂࡞ᑊࡊ࡙࡜࠹ష⏕ࡊ࡙࠷ࡾ࠾ࡢ୘᪺࡝Ⅴ࠿ኣ࠷ࠊࡱࡒິ
∸⣵⬂࡚ࡢ topoII ࠿ᵕࠍ࡝ᑻᅹን໩ࢅ♟ࡌ࠿ࠉࡐࡡ⌦⏜ࡵࢂ࠾ࡖ࡙࠷࡝࠷ࠊ⚶㐡ࡢࡆࡡࢰࣤࣂࢠ㈻ࡡᶭ⬗ࢅㄢ࡬ࡾࡡ࡞ Aspergillus nidulans ࠿᭯ຝ࡝ࡡ࡚ࡢ࡝࠷࠾࡛⩻࠻ࠉࡱࡍࡐࡡ⣵⬂හ࡚ࡡᣪິࢅ▩ࡾࡒࡴࠉ⺧කࢰࣤࣂࢠᵾㆉࢅฺ⏕ࡊࡒࣚ࢕ࣇ࢕࣒࣭ࢩࣤࢡ࡞ࡻࡽࠉtopoII ࡛ࡐࡡ௙ࡡࢰࣤࣂࢠ㈻ࡡᑻᅹࢅẒ㍉ࡊࡒࠊ
A. nidulans FGSC4 ᰬࢅ⏕࠷ࠉࢣࢿ࣑୕ࡡ topoII 㐿ఎᏄࡡ C ᮆ࡞ GFP ࢅ⤄ࡲ㎲ࢆࡓᰬࢅషᠺࡊࡒࠊኣࡂࡡ
⣵⬂࠿㟴Ḿ᭿ࡡᰶ࡛ᛦࢂࡿࡾ GFP ⺧කࢅ♟ࡌୌ᪁ࠉฦ⿛᭿࡛ᛦࢂࡿࡾ⣵⬂࡚ࡢࠉࡻࡽࢤࣤࣂࢠࢹ࡞㞗✒ࡊࡒᵕࠍ࡝ാ࠿ぜࡼࡿࡒࠊCFP ᵾㆉ histone H2A ࡡභⓆ⌟࡞ࡻࡽࠉࡐࡿࡼ࠿ᰶ㡷ᇡࡡᑻᅹ࡚࠵ࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊฦ⿛᭿ࡡᣪິࢅ㏻⤾ふᐳࡊࡒ࡛ࡆࢀࠉฦ⿛๑࡞ࡢᰶධమ࡞ୌᵕ࡞ฦᕱࡊ࡙࠷ࡒ topoII-GFP ࠿ฦ⿛᭿࡞࡝ࡾ࡛
㞗ྙࡊጙࡴࠉ々ᩐࡡᑚࡈ࡝᪺ࡾ࠷⺧ක࠿ᰶහ࡞ぜࡼࡿࡾࡻ࠹࡞࡝ࡖࡒࠊᚃ᭿௧㜾ࡢ㞗୯ࡊ࡙࠷ࡒ⺧ක࠿ᰶ㡷ᇡධమ࡫࡛᛬㏷࡞ᣉᩋࡊࡒࠊ௧୕࠾ࡼ topoII ࡢฦ⿛᭿࡞ⴥࡊ࠷ᑻᅹን໩ࢅ♟ࡌࡆ࡛࠿ࢂ࠾ࡖࡒࠊ
Cellular behavior of type II topoisomerase in Aspergillus nidulans: analysis with live cell imaging of proteinstagged with fluorescence proteins.
1

Tomohiro Akashi, 1 Mika Kawagishi, 1Akihiko Kikuchi, 2Berl R. Oakley, 3 Tetsuya Horio(1 Mol. Mycol. Med.,

Nagoya Univ. Grad. Sch.; 2 Mol. Genet., Ohio St. Univ.; 3Health Bio., Tokushima Univ. Grad. Sch.)

67

P-35
ࢨ࢕ࢰࢢ(Lentinula edodes)ࡡș-1,3-ࢡࣜ࢜ࢻ࣭ࢭࡡよᯊᆊᮇ⿩ୌ㸡Ễ஬ົ㸡୯ฝၤᏄ㸡ఫ⸠ฺḗ 㸝ᒷᡥ⏍ᕝ◂㸞ࢨ࢕ࢰࢢ(Lentinula edodes)࠾ࡼ᢫ฝ⢥⿿ࡈࡿ࡙࠷ࡾࣝࣤࢲࢻࣤࡢࠉࢨ࢕ࢰࢢࡡ⣵⬂ባࡡ୹さᵋᠺᠺฦ࡚࠵ࡾș-1,3 ࢡ࡚ࣜ࢜ࣤ࠵ࡽࠉᢘ࢝ࣤ๠࡛ࡊ࡙⮣ᗃ࡚⏕࠷ࡼࡿ࡙࠷ࡾࠊࣝࣤࢲࢻࣤࡢࠉࢨ࢕ࢰࢢಕᏋ㐛⛤࡚ exo-ࢡࣜ࢜ࢻ࣭ࢭ࡞ࡻࡽฦよࡈࡿࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊࡐࡆ࡚ᮇ◂✪࡚ࡢࠉࢨ࢕ࢰࢢ࠾ࡼ exo ࠽ࡻࡦ endo-ࢡࣜ࢜ࢻ࣭ࢭ㐿ఎᏄࢅ༟㞫ࡊࠉࢤ࣭ࢺࡈࡿ࡙࠷ࡾ㓕⣪ࡡ⢥⿿ࢅ⾔ࡖࡒࠊexg1 㐿ఎᏄࡢࠉGHF5 ࡞ᒌࡌࡾ exo-ࢡࣜ࢜ࢻ࣭ࢭ࡚࠵ࡽࠉ⏍㛏㐛⛤ࡡᯮ࡞ᙁ࠷Ⓠ⌟࠿ヾࡴࡼࡿࡒࠊࡊ࠾ࡊಕᏋ㐛⛤࡚Ⓠ⌟࠿΅ᑛࡊࠉࡱࡒࣝࣤࢲࢻࣤฦ
よὩᛮࢅᣚࡖ࡙࠷࡝࠾ࡖࡒࠊexg2 㐿ఎᏄࡢ⣊≟Ⳟ㢦࡞≁᭯ࡡ exo-ࢡࣜ࢜ࢻ࣭ࢭ࡛࢓࣐ࢿ㓗࡚ࣝ࣊ࣜ 40㸚⛤ᗐࡡ┞ྜྷᛮࢅ♟ࡊࡒࠊexg2 㐿ఎᏄࡢ⏍㛏㐛⛤ࡡᯮ࡚ᙁ࠷Ⓠ⌟ࢅ♟ࡊࡒࠊexg1, exg2 㐿ఎᏄ࡛ࡵ࡞ࠉᯮࡡఘ㛏࡞㛭ࢂࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊexg2 㐿ఎᏄࡢಕᏋ㐛⛤࡚Ⓠ⌟࠿୕᪴ࡊࠉࢗ࢘ࢪࢰࣤよᯊ࡞࠽࠷࡙ࡵࢰࣤࣂࢠ㈻ࣝ
࡚࣊ࣜࡡⓆ⌟ࡡ୕᪴࠿☔ヾࡈࡿࡒࠊࡱࡒࠉ⢥⿿ࡊࡒ EXG2 ࡢࣝࣤࢲࢻࣤฦよὩᛮࢅᣚࡖ࡙࠷ࡒࠊtlg1 㐿ఎᏄࡢࠉPR5 ࣆ࢒࣐࣭ࣛ࡞ฦ㢦ࡈࡿࡾ endo-ࢡࣜ࢜ࢻ࣭ࢭὩᛮࢅᣚࡗ᳔∸ࡡᢘⳞࢰࣤࣂࢠ㈻࡛ࠉ࢓࣐ࢿ㓗࡚ࣝ࣊ࣜ40㸚⛤ᗐࡡ┞ྜྷᛮࢅ♟ࡊࡒࠊtlg1 㐿ఎᏄࡢ᩺㩥࡝Ꮔᐁమ࡚ࡢධࡂⓆ⌟࠿ヾࡴࡼࡿࡍࠉಕᏋ㐛⛤ࡡᏄᐁమ࡚ᙁ
࠷Ⓠ⌟࠿ヾࡴࡼࡿࡒࠊࡱࡒࠉࢗ࢘ࢪࢰࣤよᯊ࡞࠽࠷࡙ࡵࢰࣤࣂࢠ㈻࡚ࣝ࣊ࣜࡡⓆ⌟ࡡ୕᪴࠿☔ヾࡈࡿࡒࠊ⢥⿿ࡊࡒ TLG1 ࡢࠉࣝࣤࢲࢻࣤฦよὩᛮࢅᣚࡖ࡙࠽ࡽࠉEXG2 ࡛࡛ࡵ࡞ಕᏋ㐛⛤࡚ࡡࣝࣤࢲࢻࣤฦよ࡞㛭ࢂࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊ
Characterization of ș-1,3-glucanases from Lentinula edodes
Yuichi Sakamoto, Masaru Nagai, Keiko, Nakade, Toshitsugu Sato
(Iwate Biotech. Res. Center)

P-36ࢨ࢕ࢰࢢࡡᏄᐁమᙟᠺ㐛⛤࡞࠽࠷࡙≁␏Ⓩ࡞Ⓠ⌟ࡌࡾ㐿ఎᏄ⩄ࡡ༟㞫࡛よᯊᐋᓧᏭᑑࠉ୯ᮟ㞖သࠉ㤷ሔᓧົᙢ㸝᲻ᯐ⥪◂㸞᭩ࡵ࣎ࣅ࣭ࣖࣚ࡝ࡀࡡࡆࡡୌࡗࢨ࢕ࢰࢢ(Lentinula edodes)࡞࠽࠷࡙ࠉcDNA-RDA(cDNA-Representational
Difference Analysis)Ἢࢅᨭንࡊࡒ㐿ఎᏄࢦࣇࢹࣚࢠࢨࣘࣤࢅモࡲࡒ⤎ᯕࠉᏄᐁమᙟᠺ㐛⛤࡞࠽࠷࡙≁␏Ⓩ࡞Ⓠ⌟ࡌࡾ 105 ࢠ࣭ࣞࣤ(Ꮔᐁమཋᇱ 51 ࢠ࣭ࣞࣤࠉᠺ⇅Ꮔᐁమ 54 ࢠ࣭ࣞࣤ)ࡡ㐿ఎᏄ cDNA ᩷∞ࡡ༟㞫࡞ᠺຉࡊࡒࠊࡆࡿࡼࡡࢠ࣭ࣞࣤࡡሲᇱ㒼าࢅよᯊࡊ஢᝷ࡈࡿࡾ࢓࣐ࢿ㓗㒼า࡞ࡗ࠷࡙ࢸ࣭ࢰ࣭࣊ࢪࢅ᳠⣬ࡊࡒ⤎ᯕࠉࢤ࣭ࢺࡈࡿࡾ⏐∸ࡢୌ⯙Ⓩ௥ㅨࠉ⣵⬂ᵋ㏸ࠉࢨࢡࢻࣜఎ㐡ࠉࢪࢹࣝࢪᚺ➽࡝࡜࡞㛭ࢂࡾࢰࣤࣂࢠ㈻࡛㧏࠷┞ྜྷᛮࢅ♟ࡌࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡽࠉᵕࠍ࡝௥ㅨ⤊㊪ࡷࢨࢡࢻࣜఎ㐡⤊㊪࠿Ꮔᐁమᙟᠺ᫤࡞഼ࡂࡵࡡ࡛⩻࠻ࡼࡿࡒࠊୌ᪁ࠉ⣑༖ᩐࡡࢠ࣭ࣞࣤࡢࢸ࣭ࢰ࣭࣊ࢪ୕ࡡ㐿ఎᏄཀྵࡦࢰࣤࣂࢠ㈻࡛ධࡂ┞ྜྷᛮ࠿ぜ࠷ࡓࡎ࡝࠾ࡖࡒࡒ
ࡴࠉࢨ࢕ࢰࢢ࡞≁᭯࡝᩺ぞ㐿ఎᏄ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡒࠊ
ࡆࡿࡼࢠ࣭ࣞࣤࡡ࠹ࡔࠉ20 ಴ࡡ㐿ఎᏄࢠ࣭ࣞࣤ࡞ࡗ࠷࡙Ꮔᐁమᙟᠺ㐛⛤࡞࠽ࡄࡾ㌹෕Ⓠ⌟ࣂࢰ࣭ࣤࢅ
RT-PCR ࡞ࡻࡽㄢ࡬ࡒ⤎ᯕࠉᏄᐁమᙟᠺ㐛⛤ࢅ㏳ࡋ࡙ᵋᠺⓏ࡞Ⓠ⌟ࡊ࡙࠷ࡾࡵࡡࠉ⬂Ꮔᙟᠺࢅ⾔࠹ᠺ⇅Ꮔᐁమ୯࡞ࡡࡲⓆ⌟ࡊ࡙࠷ࡾࡵࡡཀྵࡦᏄᐁమⓆ⏍ࡡิ᭿ṹ㝭࡚࠵ࡾᏄᐁమཋᇱ࡞ࡡࡲⓆ⌟ࡊ࡙࠷ࡾࡵࡡࠉࡡኬࡀ
ࡂฦࡄ࡙ 3 ࣂࢰ࣭ࣤ࡞ฦ㢦ࡌࡾࡆ࡛࠿ฝᮮࡒࠊࡆࡿࡼ㐿ఎᏄཀྵࡦࡐࡡ⏐∸ࢰࣤࣂࢠ㈻ࡢࢨ࢕ࢰࢢࡡᏄᐁమᙟ
ᠺ㐛⛤࡞࠽࠷࡙㔔さ࡝ᙲ๪ࢅᢰࡖ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡒࠊࡆࡿࡼ㐿ఎᏄཀྵࡦ⏐∸ࡡᶭ⬗ࢅㄢ࡬ࡾࡆ࡛࡞ࡻࡽࠉ
ࢨ࢕ࢰࢢࡡࡲ࡝ࡼࡍࡀࡡࡆ㢦ධ⯙ࡡᏄᐁమᙟᠺࡡ࣒࢜ࢼࢫ࣑ࡡよ᪺࠿᭿ᙽࡈࡿࡾࠊ
Molecular cloning of developmentally specific genes by representational difference analysis during the fruiting
body formation in the basidiomycete Lentinula edodes
Yasumasa Miyazaki, Masaya Nakamura, Katsuhiko Babasaki(Forestry and Forest Products Research Institute (FFPRI))

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ᢰᏄⳞ Phanerochaete chrysosporium ࠿᭯ࡌ ࡾ P450 㐿ఎᏄࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣛࣤࢡ
୯ᮟ▩ᜠ 1ࠉᗽᮆួႵ 2ࠉᖲሪᐃ༡ 1 ࠉᒜ⏛ಞྒྷ 3ࠉばἡᨳ஦୔ 3 ࠉ᭯Ἁ ❮ 2ࠉᜇᕖ ༡ 2ࠉ๪▴༡஄ 1
㸝1 ஐኬ㝌㎨ࠉ2 ࣒ࣜࢨࣔࣤ⏍㈠◂ࠉ3JST ๪▴ࣈࣞࢩ࢘ࢠࢹ㸞࠘⥬ゕ࠙㎾ᖳࠉⓉⰅ⭁ᮑⳞ Phanerochaete chrysosporium ࡡධሲᇱ㒼า࠿ප㛜ࡈࡿࠉ࢓ࢿࢷ࣭ࢨࣘࣤࡡ⤎ᯕ࠾
ࡼ 154 ࡡ P450 ࠿Ꮛᅹࡌࡾࡆ࡛࠿♟ࡈࡿࡒࠊࡆࡿࡱ࡚࡞᪜▩ࡡ P450 ࡡ௙࡞ࠉᢰᏄⳞ࡞≺≁࡝ⰶ㤮᪐໩ྙ∸ฦよ࡞㛭୙ࡌࡾ P450 ࡡᏋᅹ࠿᭿ᙽࡈࡿ࡙࠷ࡾࠊࡈࡼ࡞ࠉỀ㓗໩ཬᚺ࡝࡜ࢅり፳ࡌࡾ P450 ࡢࠉ⏐ᴏ⏲࡚ࡵይ㐲࡝り፳ࢵ࣭࡛ࣜ࡝ࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉP. chrysosporium P450 ࡡᶭ⬗ࡢ࡮࡛ࢆ࡜᪺ࡼ࠾࡞࡝ࡖ࡙࠷࡝
࠷ࠊࡐࡆ࡚ᮇ◂✪࡚ࡢࠉ࣏࢕ࢠࣞ࢓ࣝ࢕Ἢࢅ⏕࠷ࠉᵾⓏ໩ྙ∸ࡇ࡛࡞ㄇᑙⓏ࡞Ⓠ⌟ࡌࡾ P450 ࡡ᥀⣬ࢅモࡲࡒࠊ࠘ᡥἪ࠽ࡻࡦ⤎ᯕ࠙P. chrysosporium ⏜ᮮࡡᏰධ㛏 cDNA 132 ⛸ࢅࣈ࣭ࣞࣇ࡛ࡊ࡙⏕࠷ࡒࠊHCLN ᮪௲ୖ࡚ 3᪝㛣ᇰ㣬ᚃࠉ⛸ࠍࡡ໩ྙ∸ࢅ῟ຊࡊࠉᇰ㣬ࢅ⤽⤾ࡊࡒࠊᡜᏽ᫤㛣ᚃࠉᚋࡼࡿࡒⳞమ࠾ࡼ mRNA ࢅ᢫ฝࡊࠉ㏣㌹෕ཬᚺࢅ⾔ࡖࡒࠊࢤࣤࢹ࣭ࣞࣜࠉ໩ྙ∸῟ຊୖࡻࡽᚋࡼࡿࡒ cDNA ࢅࡐࡿࡑࡿ Cy3, Cy5 ࡚⺧කࣚ࣊ࣜࡊ࡙
࣏࢕ࢠࣞ࢓ࣝ࢕よᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉࢺࢸ࢜ࢿ࣭ࣜ῟ຊ࣬ࣁࢼࣛࣤ῟ຊ࣬Ꮽ᜝㤮㓗῟ຊ࡞ࡻࡽࢤࣤࢹࣞ
࣭࡛ࣜẒ࡬࡙ 2 ಶ௧୕ࡡⓆ⌟ࢅ♟ࡊࡒ P450 ࠿ᩐ⛸ぜ࠷ࡓࡈࡿࡒࠊ⌟ᅹࠉࡈࡼ࡞レ⣵࡞よᯊࢅ⾔ࡖ࡙࠷ࡾࠊ

Microarray profiling of cytochrome P450 gene expression in the white-rot fungus Phanerochaete chrysosporium
Chie Nakamura1, Shinji Hirosue2, Nobuhiro Hiratsuka1, Shuji Yamada3, Masafumi Oyadomari3, Akira Arisawa2,
Hiroshi Tsunekawa2, Hiroyuki Wariishi1 (1Faculty of Agriculture, Kyushu Univ., 2Bioresource Laboratories, Mercian
Co. 3JST)P-38
ᢰᏄⳞ Phanerochaete chrysosporium ࡞࠽ࡄ ࡾࢬ࣭ࣜࣞࢪฦよ㓕⣪㐿ఎᏄ
Ⓠ⌟ᚺ➽࡞㛭ࡌࡾ◂✪㕝ᮄୌྍࠉ஫༎ᔋᅽ᪝Ꮔࠉ㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞 ᮄᮞ⭁ᮑᢰᏄⳞ Phanerochaete chrysosporium ࡡࢬ࣭ࣜࣞࢪฦよ࡞࠽ࡄࡾᇱ㈻ࡡヾㆉᶭᵋࡢᮅࡓ୘᪺࡝Ⅴ࠿ኣ࠷ࠊࡆࡿࢅ᪺ࡼ࠾࡞ࡌࡾࡒࡴࠉᇰᆀᠺฦ࡛ࡊ࡙ࡡ⅛⣪″ࡡ⛸㢦ࡷ⃨ᗐࠉ࠵ࡾ࠷ࡢ✽⣪⃨ᗐ࠿ࢬ࣭ࣜࣞࢪฦ
よ࡞㛭㏻ࡌࡾ㓕⣪㐿ఎᏄࡡⓆ⌟ᚺ➽࡞࡜ࡡࡻ࠹࡞ᙫ㡢ࡌࡾ࠾࡞ࡗ࠷࡙よᯊࢅ⾔ࡖࡒࠊ P. chrysosporium ࢅ✽⣪⃨ᗐ࠽ࡻࡦ⅛⣪″ࡡ⛸㢦ࠉ⃨ᗐࡡ␏࡝ࡾᇰᆀࢅ⏕࠷࡙ 3 ᪝㛣ᇰ㣬ࡊࠉⳞమ࠾ࡼ mRNAࢅ᢫ฝࡊࠉRT-PCR ࡞ࡻࡽ᪜▩ࡡࢬ࣭ࣜࣞࢪฦよ㛭㏻㓕⣪㐿ఎᏄࡡⓆ⌟ᚺ➽よᯊࢅ⾔ࡖࡒࠊ⅛⣪″࡞ࡢࢬ࣭ࣜࣞࢪࠉࢡࣜࢤ࣭ࢪࠉࢬࣞࣄ࢛࣭ࢪࢅࡐࡿࡑࡿ⏕࠷ࡒࠊࡐࡡ⤎ᯕࠉ྘㐿ఎᏄⓆ⌟㔖ࡡ✽⣪⃨ᗐ࡞ࡻࡾᕣࡢ࡮࡛ࢆ࡜ぜࡼࡿ࡝࠾ࡖࡒࠊୌ᪁ࠉ⅛⣪″ࡡ⛸㢦࠽ࡻࡦ⃨ᗐࡡᙫ㡢࡞ࡗ࠷࡙ࡢࠉ3 ᪝㛣ࡡᇰ㣬࡚ࡢ 0.02%ࡡࢬࣞࣄ
࢛࣭ࢪ࡞࠽࠷࡙࡮࡛ࢆ࡜ࡡ㐿ఎᏄⓆ⌟࠿ቌຊࡊࡒࠊࢡࣜࢤ࣭ࢪࢅ⅛⣪″࡛ࡊ࡙⏕࠷ࡒᇰ㣬⣌࡞࠽࠷࡙ࡵࠉ఩
⃨ᗐࡡ࡛ࡀ࡞Ⓠ⌟ࣝ࣊ࣜ࠿ቌຊࡌࡾ㐿ఎᏄ࠿ኣ࠾ࡖࡒࠊḗ࡞ࢡࣜࢤ࣭ࢪࢅ⏕࠷࡙ 3 ᪝㛣๑ᇰ㣬ࡊࡒⳞమࢅ⅛⣪″ࢅྱࡱ࡝࠷ᇰᆀ࡚ 6 ᫤㛣ᇰ㣬ࡊࡒࠊࡐࡡᚃ྘⢶ࢅ῟ຊࡊ࡙ࡈࡼ࡞ 6 ᫤㛣ᇰ㣬ࡊྜྷᵕ࡞Ⓠ⌟ᚺ➽ࢅよᯊࡊࡒࠊࡐࡡ⤎ᯕࠉࢬࣞࣄ࢛࣭ࢪ⃨ᗐ࠿㧏࠷࡛ࡀ࡞Ⓠ⌟㔖࠿ቌຊࡌࡾ㐿ఎᏄ࠿ኣࡂࠉࡱࡒࢡࣜࢤ࣭ࢪ⃨ᗐ࠿఩࠷࡛ࡀ࡞ࡢࢡࣜࢤ࣭ࢪ࡞ࡻࡾⓆ⌟ᢒโࡢࡲࡼࡿࡍ࡞Ⓠ⌟㔖࠿ቌຊࡊࠉ⃨ᗐ࠿㧏ࡂ࡝ࡾ࡞ࡗࡿⓆ⌟㔖࠿΅ᑛࡌࡾലྡྷ࠿ࡲࡼࡿࡒࠊ῟ຊࡊࡒ⢶࠿఩⃨ᗐࡡሔྙ࡚ࡢࢬࣞࣄ࢛࣭ࢪࡻࡽࡵࢡࣜࢤ࣭ࢪࢅ⏕࠷ࡒሔྙ࡞Ⓠ⌟㔖ࡡቌຊ࠿ⴥࡊ࠾ࡖࡒࡆ࡛࠾ࡼࠉ⢶⃨ᗐ࠿఩࠷࡛ࡀ࡞ࡢࢡࣜࢤ࣭ࢪ࠿ࢬ࣭ࣜࣞࢪฦよࡡᘤࡀ㔘࡛࡝ࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊ
Analysis of gene expression involved in cellulose degradation by basidiomycete Phanerochaete chrysosporium
Hitoshi Suzuki, Kiyohiko Igarashi, Masahiro Samejima (Univ. of Tokyo)

69

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ᢰᏄⳞ Phanerochaete chrysosporium ⏜ᮮࢹ ࣝࣀ࣭ࣞࢪຊࣛࣤ㓗ฦよ㓕⣪ࡡᶭ⬗ よᯊᖲ▴ḿ⏠ࠉ஫༎ᔋᅽ᪝Ꮔࠉ㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞 ࢬ࣭ࣜࣞࢪࡢᆀ⌣୕࡚᭩ࡵ㇇ᐣ࡞Ꮛᅹࡌࡾࣁ࢕࢛࣏ࢪ࡚࠵ࡾࡆ࡛࠾ࡼࠉ௛ຊ౮ೋࡡ㧏࠷᭯⏕⢶㈻࡫ࡡንᥦ
ᢇ⾙ࡡ㛜Ⓠ࠿᭻ࡱࡿ࡙࠷ࡾࠊᮇ◂✪࡚ࡢࠉ㓕⣪ࢅ⏕࠷ࡒࢬ࣭ࣜࣞࢪࡡࢹࣝࣀ࣭ࣞࢪ࡫ࡡንᥦࢅ┘ᣞࡊ࡙ࠉᢰ
ᏄⳞ Phanerochaete chrysosporium ⏜ᮮࢹࣝࣀ࣭ࣞࢪຊࣛࣤ㓗ฦよ㓕⣪(TP)ࡡᶭ⬗ࢅ᪺ࡼ࠾࡞ࡌࡾࡆ࡛ࢅ┘Ⓩ࡛ࡊࡒࠊ P. chrysosporium ࡡࢣࢿ࣑ࢸ࣭ࢰ࣭࣊ࢪࢅฺ⏕ࡊ࡙ TP ᵕ㐿ఎᏄࢅࢠ࣭ࣞࢼࣤࢡࡊࠉሲᇱ㒼าࢅỬᏽࡊࡒࠊࡆࡡ㐿ఎᏄࢅ⏕࠷࡙ኬ⭘Ⳟࢅᙟ㈻㌹ᥦࡊࠉ⤄ᥦ࠻ࢰࣤࣂࢠ㈻ࡡ⏍⏐࠽ࡻࡦ⢥⿿ࢅ⾔ࡖࡒࠊ⢥⿿ࡊࡒ⤄ᥦ࠻ࢰ
ࣤࣂࢠ㈻ࢅࢡࣜࢤ࣭ࢪ࠽ࡻࡦࢡࣜࢤ࣭ࢪ 1-ࣛࣤ㓗࡛࢕࣭ࣤ࢞ࣖ࣊ࢹࡊࠉཬᚺ⏍ᠺ∸ࢅ⷟ᒒࢠ࣏ࣞࢹࢡࣚࣆ࢔
࣭㸝TLC㸞࡞ࡻࡖ࡙よᯊࡊࡒࠊ ᮇᐁ㥺࡚ࢠ࣭ࣞࢼࣤࢡࡈࡿࡒ TP ᵕ㐿ఎᏄࡢࠉࢡࣛࢤࢨࣜࢹࣚࣤࢪࣆ࣭࢘ࣚࢭࢡ࣭ࣜࣈ 1 ࡡ࣓ࢲ࣭ࣆࢅᣚࡗࠉ
⢶㌹⛛㓕⣪ࣆ࢒࣐࣭ࣛ4 ࡞ᒌࡌࡾࢰࣤࣂࢠ㈻ࢅࢤ࣭ࢺࡊ࡙࠷ࡾࡆ࡛࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊ⢥⿿ࡊࡒ⤄ᥦ࠻ࢰࣤ
ࣂࢠ㈻ࢅ SDS-PAGE ࡞౩ࡊࡒ࡛ࡆࢀࠉฦᏄ㔖 80kDa ௛㎾࡞༟ୌࡡࣁࣤࢺ࠿☔ヾࡈࡿࡒࠊࡱࡒࠉTLC ฦᯊࡡ⤎ᯕࠉࢹࣝࣀ࣭ࣞࢪᵾဗ࡛ྜྷࡋ Rf ೋࢅ୙࠻ࡾࢪ࣎ࢴࢹ࠿☔ヾ࡚ࡀࡒࡆ࡛࠾ࡼࠉᮇ㐿ఎᏄ࠿ P. chrysosporium ⏜ᮮ TP ࢅࢤ࣭ࢺࡌࡾࡵࡡ࡚࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ⌟ᅹࠉ⤄ࡲᥦ࠻ TP ࡡᶭ⬗よᯊࢅ⾔ࡖ࡙࠷ࡾࠊ
Characterization of a trehalose phosphorylase from the basidiomycete
Phanerochaete chrysosporium
Masao Hiraishi, Kiyohiko Igarashi,Masahiro Samejima (Univ. Tokyo)

P-40
ᢰᏄⳞ Phanerochaete chrysosporium ࠿⏍⏐ࡌࡾ⢶㈻ຊỀฦよ㓕⣪ࣆ࢒࣐࣭ࣛ74 ࢞ࢨ ࣞࢡࣜ࢜ࢻ࣭ࢭࡡᶭ⬗よᯊ▴⏛༜ஒ 1ࠉຊ἖ఫᖲ 1ࠉ஫༎ᔋᅽ᪝Ꮔ 1ࠉ▦㏛ග㑳 2ࠉ᪝ྚ࠵ࡷᏄ 2ࠉ୔▴ Ꮽ 2ࠉ㩢ᓞḿᾀ 1
㸝1 ᮶ኬ㝌࣬㎨⏍⛁ࠉ2 ⏐⥪◂㸞ᢰᏄⳞ Phanerochaete chrysosporium ࡢࠉ᳔∸⣵⬂ባࢅᵋᠺࡌࡾኣ⢶ࢅฦよࡌࡾ㐛⛤࡚ᵕࠍ࡝⢶㈻ຊỀฦよ㓕⣪㸝GH㸞ࢅⳞమአ࡞⏍⏐ࡌࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊࡊ࠾ࡊ࡝࠿ࡼࠉࡐࡿࡼࡡ࠹ࡔ㓕⣪ᏕⓏ࡝࢞ࣔࣚࢠࢰࣚ
࢕ࢫ࠿⾔ࢂࡿࡒࡵࡡࡢࢂࡍ࠾࡚࠵ࡽࠉኣࡂࡡⳞమአ GH ࡡᶭ⬗࠿ᮅ▩ࡡࡱࡱ࡚࠵ࡾࠊࡐࡆ࡚ᠻࠍࡢࠉࢣࢿ࣑
᝗ሒࢅฺ⏕ࡊ࡙ࠉ⣵⬂ባኣ⢶ฦよ㐛⛤࡚⏍⏐ࡈࡿࡾⳞమአࢰࣤࣂࢠ㈻ࡡࣈࣞࢷ࢛࣭࣑よᯊࢅ⾔࠷ࠉࡐࡡ㐛⛤࡚ᮇⳞ࠿ࢬ࣭ࣜࣞࢪᇰ㣬⣌࡞࠽࠷࡙ GH ࣆ࢒࣐࣭ࣛ74 ࡞ᒌࡌࡾᮅ▩ࢰࣤࣂࢠ㈻ࢅ⏍⏐ࡊ࡙࠷ࡾࡆ࡛ࢅ᪺ࡼ࡞
ࡊ࡙ࡀࡒࠊࡐࡆ࡚ᮇ◂✪࡚ࡢࠉGH ࣆ࢒࣐࣭ࣛ74 ࡞ᒌࡌࡾ㓕⣪ࢅࢤ࣭ࢺࡌࡾ cDNA ࡡሲᇱ㒼าࢅỬᏽࡌࡾ࡛࡛ࡵ࡞ࠉ࣒ࢰࢿ࣭ࣜ㈠໩ᛮ㓕ẍࢅ⏕࠷࡙⤄ᥦ࠻㓕⣪ࢅ⏍⏐ࡊࠉᮇⳞ⏜ᮮࡡ GH ࣆ࢒࣐࣭ࣛ74 ࡞ᒌࡌࡾ㓕⣪ࡡ࢞ࣔࣚࢠࢰࣚ࢕ࢫࢅ⾔ࡖࡒࠊⓆ⌟ࡈࡎࡒ⤄ᥦ࠻㓕⣪ࢅࢰ࣏ࣛࣤࢺ⏜ᮮࡡ࢞ࢨࣞࢡࣜ࢜ࣤ࡞ష⏕ࡈࡎࡒ࡛ࡆࢀࠉ
࢞ࢨࣞࢡ࢛ࣜ࢜ࣤࣛࢥ⢶ࢅ⏍ᠺ∸࡛ࡊ࡙୙࠻ࡒࡆ࡛࠾ࡼࠉᮇⳞ⏜ᮮࡡ GH ࣆ࢒࣐࣭ࣛ74 ࡞ᒌࡌࡾ㓕⣪࠿࢞ࢨࣞࢡࣜ࢜ࢻ࣭ࢭ࡚࠵ࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ⌟ᅹࠉᵕࠍ࡝ᇱ㈻ࢅ⏕࠷࡙ᮇ㓕⣪ࡡཬᚺᛮࢅㄢ࡬࡙࠷ࡾࠊ
Characterization of the glycoside hydrolase family 74 xyloglucanase from the basidiomycete Phanerachaetechrysosposrium
Takuya Ishida1, Taira Kajisa1, Kiyohiko Igarashi1, Katsuro Yaoi2, Ayako Hiyoshi2, Yasushi Mitsuishi2, Masahiro
Samejima1 (1Univ. of Tokyo, 2AIST)

70

P-41
ᢰᏄⳞ Phanerochaete chrysosporium ࠿⏍⏐ ࡌࡾ⢶㈻ຊỀฦよ㓕⣪ࣆ࢒࣐࣭ࣛ1 ࡞ᒌࡌࡾș-ࢡࣜࢤࢨࢱ࣭ࢭࡡᶭ⬗よᯊሪ⏛๙ኃ 1ࠉ஦༎㌲᜾㔓 1ࠉྚ⏛ㄌ 2 ࠉఄಘ㐅▦ 1ࠉ஫༎ᔋᅽ᪝Ꮔ 1ࠉ㩢ᓞḿᾀ 1㸝1 ᮶ኬ㝌࣬㎨⏍⛁ࠉ2 㣏⥪◂㸞ࢬ࣭ࣜࣞࢪࡡ㓕⣪⢶໩ࡢࠉࢬ࣭ࣜࣚࢭ࡞ࡻࡖ࡙วࡽฝࡈࡿ࡙ࡀࡒࢬࣞࣄ࢛࣭ࢪࡷࢬ࢛ࣞࣛࢥ⢶ࢅࠉș-ࢡࣜࢤࢨࢱ࣭ࢭ㸝BGL㸞࠿ࢡࣜࢤ࣭ࢪ࡫࡛ንᥦࡌࡾࡆ࡛࡚⾔ࢂࡿࡾࠊࡆࡿࡱ࡚ࢬ࣭ࣜࣚࢭ࡞ࡗ࠷࡙ࡢኣࡂࡡ◂✪࠿Ⅵࡈࡿ࡙࠷ࡾࡡ࡞ᑊࡊࠉș-ࢡࣜࢤࢨࢱ࣭ࢭ㸝BGL㸞࡞ࡗ࠷࡙ࡢ◂✪ౚ࠿ᑛ࡝ࡂࠉຝ⋙Ⓩ࡝ BGL ࡢᚋࡼࡿ࡙࠷࡝࠷ࠊᮇ◂✪ᐄ࡚ࡢࠉᢰᏄⳞ Phanerochaete chrysosporium ࠿⏍⏐ࡌࡾ⢶㈻ຊỀฦよ㓕⣪ࣆ࢒࣐࣭ࣛ㸝GHF㸞
3 ࡞ฦ㢦ࡈࡿࡾ BGL㸝BGL3㸞ࢅㄢ࡬࡙ࡀࡒ࠿ࠉࡆࡡ BGL ࡢࢬࣞࣄ࢛࣭ࢪ࡞ᑊࡌࡾཬᚺᛮ࠿இࡊ࠷ࡆ࡛࠿᪺
ࡼ࠾࡛࡝ࡖ࡙࠷ࡾࠊP. chrysosporium ࢣࢿ࣑᝗ሒ࡞ࡻࡾ࡛ࠉGHF1 ࡞ᒌࡌࡾ BGL ᵕ㐿ఎᏄ࠿ 2 ࡗᏋᅹࡌࡾࡆ࡛
࠿᪺ࡼ࠾࡛࡝ࡖ࡙࠷ࡾࡒࡴࠉᮇ◂✪࡚ࡢࡐࡿࡼࡡ㐿ఎᏄ࠿ࢤ࣭ࢺࡌࡾ cDNA ࢅࢠ࣭ࣞࢼࣤࢡࡊࠉ␏⛸Ⓠ⌟⣌
࡞ࡻࡽ⤄ᥦ࠻మ࡛ࡊ࡙⏍⏐ࡊᶭ⬗よᯊࢅ⾔ࡖࡒࠊᮇ◂✪࡚ࢠ࣭ࣞࢼࣤࢡࡈࡿࡒ 2 ࡗࡡ cDNA ࠿ࢤ࣭ࢺࡊ࡙࠷ࡾࢰࣤࣂࢠ㈻㸝BGL1AࠉBGL1B㸞ࡡ᥆ᏽ࢓࣐ࢿ㓗㒼าࢅよᯊࡊࡒ⤎ᯕࠉ୦⩽ࡢ 65%ࡡ┞ྜྷᛮࢅ♟ࡊࡒࡆ࡛࠾ࡼப࠷࡞㢦జࡊࡒᵋ㏸ࢅ᭯ࡌࡾྊ⬗ᛮ࠿♟ြࡈࡿࡒࠊ୦ BGL ࡡ pH ౪Ꮛᛮࢅῼᏽࡊࡒ࡛ࡆࢀࠉ
࡜ࡔࡼࡵ pH6.5 ࡚᭩ኬὩᛮࢅ♟ࡊࡒ࠿ࠉ⮾࿝῕࠷ࡆ࡛࡞ࠉBGL1Bࡢ㓗ᛮഁ࡚࡮࡛ࢆ࡜Ὡᛮࢅ♟ࡈ࡝࠾ࡖࡒࡡ࡞ᑊࡊ࡙ࠉBGL1A ࡢ pH4.0 ࡞࠽࠷࡙ࡵ᭩ኬὩᛮࡡ 50%௧୕ࡡὩᛮ
ࢅಕᣚࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊࡈࡼ࡞ࠉࢬࣞࣄ࢛࣭ࢪ࡞ᑊࡌࡾཬᚺ㏷ᗐࣂ࣒࣭ࣚࢰࢅ⟤ฝࡊࡒ࡛ࡆࢀࠉ
BGL1B ࡢ BGL1A ࡷ BGL3 ࡛Ẓ㍉ࡊ࡙ࢬࣞࣄ࢛࣭ࢪ࡞ᑊࡌࡾば࿰ᛮ࠿㧏ࡂࠉ᭩ࡵ㧏࠷り፳ຝ⋙ࢅ♟ࡊࡒࡆ࡛࠾ࡼࠉBGL1B ࠿ࢬ࣭ࣜࣞࢪࡡ⢶໩࡞㐲ࡊࡒ㓕⣪࡚࠵ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
Characterization of glycoside hydrolase family 1 ș -glucosidases from the basidiomycete Phanerochaetechrysosporium
Takeshi Tsukada1, Yuri Nijikken1, Makoto Yoshida1, 2, Shinya Fushinobu1, Kiyohiko Igarashi1, Masahiro Samejima1
㸝1Univ. of Tokyo, 2Natl. Food Res. Inst㸞

P-42
DNA ࡡ㟸≁␏Ⓩቌᖕࢅฺ⏕ࡊࡒᮄᮞ⭁ᮑ Ⳟࡡ㧏វᗐ᳠ฝ࿰⏛᭰Ꮔ㸡ຊ἖ఫᖲ㸡஫༎ᔋᅽ᪝Ꮔ㸡㩢ᓞḿᾀ㸝᮶ኬ㝌࣬㎨⏍⛁㸞ᮄᮞ⭁ᮑⳞࡢᮄ㈻ᵋ㏸∸ࡡᙁᗐ຋໩ࢅᘤࡀ㉫ࡆࡊ㸡ᆀ㟀࡞ࡻࡾᮄ㈻పᏩࡡ◒᥾ࡷ಻ቪࡡኬࡀ࡝ཋᅄࡡࡥ࡛
ࡗ࡛ࡈࡿ࡙࠷ࡾࡆ࡛࠾ࡼ㸡ప⎌ሾ࠽ࡄࡾᮄᮞ⭁ᮑⳞࡡິឺࢅ࣓ࢼࢰࣛࣤࢡࡊ࡙࠷ࡂࡆ࡛ࡢ㸡ᮄ㈻పᏩࡡ⥌ᣚ⟮⌦ࡡࡒࡴ࡞㔔さ࡝ㄚ㢗࡛ゕ࠻ࡾࠊᮄᮞ⭁ᮑⳞࡡኣࡂࡢᢰᏄⳞ࡞ฦ㢦ࡈࡿࡾ࠿㸡ࡆࡿࡱ࡚ࡢ㸡⭁ᮑᮞ࠾ࡼ᤿
ཱིࡊࡒⳞࢅᇰ㣬ࡊ㸡ࡐࡡᙟឺࢅふᐳࡌࡾࡆ࡛࡚Ⳟࡡྜྷᏽࢅ⾔ࡖ࡙ࡀࡒࠊࡊ࠾ࡊ࡝࠿ࡼ㸡ࡆࡡᡥἪࡢᮅ▩ࡡ᭯ᐐⳞࢅᇰ㣬ࡊ࡙ࡊࡱ࠹༱㝜ᛮࡷ᧧షࡡ↱㞟ࡈ࠿ᣞᦤࡈࡿ࡙࠷ࡾࠊࡐࡆ࡚᭩㎾࡞࡝ࡖ࡙㸡ᢰᏄⳞࡡ᳠ฝ࡛ྜྷᏽ
ࢅ┘Ⓩ࡛ࡊ࡙㸡rDNA ࡡ ITS 㡷ᇡࢅ PCR ࡞ࡻࡖ࡙ቌᖕࡌࡾᡥἪ࠿㛜Ⓠࡈࡿࡒ࠿㸡ᮄᮞ࡞ྱࡱࡿࡾᠺฦ࠿ PCR
ࢅ㜴ᐐࡊࡒࡽ㸡㗢ᆵ DNA ࢅᚋࡾࡒࡴ࡞ኣ㔖ࡡࢦࣤࣈࣜࢅᚪさ࡛ࡌࡾࡒࡴ㸡⌟ᐁⓏ࡝࣓ࢼࢰࣛࣤࢡἪ࡛ࡊ࡙ࡢ
ၡ㢗࠿࠵ࡾࠊࡐࡆ࡚ᮇ◂✪࡚ࡢ㸡Phi29 DNA ࣒࣭࣎ࣛࣚࢭ࡛ࣚࣤࢱ࣑ࣉ࢞ࢦ࣏࣭ࢅ⏕࠷ࡒ DNA ቌᖕἪࢅ᪜Ꮛ
ࡡ᪁Ἢ࡞⤄ࡲྙࢂࡎࡾࡆ࡛࡚㸡᚜㔖ࢦࣤࣈࣜ࠾ࡼࡡⳞᰬྜྷᏽ࡛Ẓ㍉ᏽ㔖ࢅྊ⬗࡛ࡌࡾᡥἪࡡ㛜Ⓠࢅモࡲࡒࠊ྘⛸⣊≟ⳞࡡⳞ⣊ࢅࣁࢴࣆ࢒࣭୯࡚◒◃ࡊࡒᚃ㸡ࢦࣤࣈࣜ୯ࡡධ࡙ࡡ DNA ࢅ୕オࡡ᪁Ἢ࡞ࡻࡽቌᖕࡊࡒࠊ
ḗ࡞㸡ᚋࡼࡿࡒ DNA ࢅ㗢ᆵ࡛ࡊ࡙㸡rDNA ࡡ ITS 㡷ᇡࢅቌᖕࡌࡾࡒࡴࡡ⣊≟Ⳟ࡞≁␏Ⓩ࡝ࣈࣚ࢕࣏࣭㸡࠽ࡻࡦᢰᏄⳞ࡞≁␏Ⓩ࡝ࣈࣚ࢕࣏࣭ࢅ⏕࠷࡙ PCR ࢅ⾔࠷㸡ᏄᄙⳞ࡛ᢰᏄⳞ࠿ฦ㞫࡚ࡀࡾࡆ࡛ࢅ☔ヾࡊࡒࠊࡈࡼ࡞㸡々ᩐ⛸ࡡⳞࢅᵕࠍ࡝Ẓ⋙࡞Ίྙࡊࡒࢦࣤࣈࣜࢅ⏕࠷࡙ྜྷᵕࡡᡥ㡨࡚ PCR ࢅ⾔ࡖࡒ࡛ࡆࢀ㸡ࡐࡿࡑࡿࡡⳞ⏜ᮮࡡ ITS 㡷ᇡ࠿ࢦࣤࣈࣜ୯ࡡⳞࡡΊྙẒࢅཬ᫆ࡊ࡝࠿ࡼቌᖕࡊࡒࠊ௧୕ࡡ⤎ᯕ࠾ࡼ㸡᚜㔖ࡡ⭁ᮑᮄᮞモ㥺మࢅᐁ㥺࡞౩ࡊࡒሔྙ࡚ࡵ㸡⭁ᮑⳞࡡᏋᅹ☔ヾ࠽ࡻࡦࡐࡡྜྷᏽ㸡ࡈࡼ࡞Ẓ㍉ᏽ㔖࡚ࡀࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ
Sensitive detection of wood-rotting fungi using non-specific amplification of DNA
Tomoko Wada, Taira Kajisa, Kiyohiko Igarashi, Masahiro Samejima (Univ. of Tokyo)

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㯔Ⳟ࡞࠽ࡄࡾ᩺ぞࡡ⢶㙈㛭㏻Ὡᛮࢰࣤࣂࢠࡡ᥀⣬⋚㔕Ꮟୌ㸡ᑈᯐ㟱ᐃ㸡ఫ⸠ཪ⣎㸡ᇷ㉲Ờ⨶*㸡▹ᕖ⨶ఫ⥬㸡ᒜ᰷೒Ꮔ㸡⏣⏛㞖஄㸝⏐⥪◂㹺⏍∸ᶭ⬗㸡*᪝ኬ࣬⏍⏐ᕝᏕ㸞㯔Ⳟ࠿⏍⏐ࡌࡾࢰࣤࣂࢠࡡ୯࡚⢶㙈࡞ᑊࡊ࡙ฦよ࠵ࡾ࠷ࡢ⤎ྙࡡὩᛮࢅ᭯ࡌࡾ᩺ぞࡡ⏐ᴏ୕᭯⏕࡝ࢰࣤࣂࢠࡡ᥀⣬ࢅ⾔ࡖ࡙࠷ࡾࠊ᩺ぞࢰࣤࣂࢠࡡ᥀⣬ࡢ⿿ဗフ౮ᢇ⾙ᇱ┑ᶭᵋ(NITE)࡛ࡡභྜྷ◂✪࡚⏐Ꮥᏻ㏻ᦘࡡࢤ
ࣤࢮ࣭ࢨ࢓࣑࡞࠽࠷࡙⾔ࢂࡿࡒ㯔Ⳟࡡࢣࢿ࣑よᯊࡡ⤎ᯕ࡞ᑊࡌࡾ࣓࣌ࣞࢩ࣭よᯊ࡞ࡻࡽ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉ஦⛸㢦ࡡ᩺ぞࡡࢡࣜ࢜ࢻ࣭ࢭು⿭㐿ఎᏄ࡛ᄿ⛸㢦ࡡ᩺ぞࡡࣝࢠࢲࣤು⿭㐿ఎᏄ࡛ࢅぜࡗࡄฝࡊࡒࠊࡆࡿࡼࡡ
㐿ఎᏄࡡ cDNA ࢅ㯔Ⳟࡻࡽష⿿ࡊࡒ cDNA ࣚ࢕ࣇ࣭ࣚࣛࡻࡽ PCR ࡞࡙ቌᖕࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉࡆࡿࡼࡡ㐿ఎᏄࡢࢣࢿ࣑よᯊ࡚஢ῼࡈࡿࡒࡵࡡ࡛࡮࡯ྜྷࡋኬࡀࡈ࡚ቌᖕࡈࡿࡒࠊቌᖕࡈࡿࡒ㐿ఎᏄࡡโ㝀㓕⣪ฌ⌦ᚃࡡ⏍ᠺ DNA ᩷∞ࡡ࢓࣭࢝ࣞࢪࢣࣜ㞹Ẵὃິ࡚ࡡࣁࣤࢺࣂࢰ࣭ࣤࡡよᯊ࡞ࡻࡽࠉ྘㐿ఎᏄࡡࢪࣈࣚ࢕ࢨࣤࢡ㒂న
ࡵࢣࢿ࣑よᯊ࡚஢ῼࡈࡿࡒࡵࡡ࡛࡮࡯ྜྷࡋ࡚࠵ࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
ḗ࡞ࠉࡆࡿࡼࡡቌᖕࡊࡒ྘ cDNA ࢅኬ⭘Ⳟኬ㔖Ⓠ⌟࣊ࢠࢰ࣭࡚࠵ࡾ pET21b ࡞ 6His ࢰࢡ࠿௛ຊࡈࡿࡒ⼝ྙࢰࣤࣂࢠ࡛ࡊ࡙Ⓠ⌟ࡈࡿࡾࡻ࠹࡞ࢠ࣭ࣞࢼࣤࢡࡊ࡙ࠉኬ⭘Ⳟ࡞࠽࠷࡙ኬ㔖Ⓠ⌟ࢅモࡲࡒࠊࡐࡡ⤎ᯕࠉୌ⛸㢦ࡡࢡࣜ࢜ࢻ࣭ࢭು⿭࡛ୌ⛸㢦ࡡࣝࢠࢲࣤು⿭ࡡⓆ⌟࠿☔ヾࡈࡿࡒࠊ௑ᚃࠉⓆ⌟ࡡヾࡴࡼࡿࡒࢰࣤࣂࢠ࡞㛭ࡊ࡙ࢼࢴࢢ࣑ࣜ࢜ࣚ࡞࡙⢥⿿ᚃࠉὩᛮῼᏽࠉᇱ㈻≁␏ᛮ࣬⪇⇍ᛮ࡝࡜ࡡᶭ⬗よᯊ࠽ࡻࡦㅎᛮ㈻ࡡ᳠ゞࢅ⾔࠹஢ᏽ࡚࠵ࡾࠊ
Expression and characterization of novel proteins of Aspergillus oryzae that have activities for degradation of orbinding to carbohydrate chains
Koichi Tamano, Yasunobu Terabayashi, Yuki Satou, Emi Horikoshi*, Misao Sunagawa, Noriko Yamane, Masayuki
Machida 㸝AIST, *Nihon Univ.㸞

P-44
A. oryzae ࡞࠽ࡄࡾ N-⤎ྙᆵ⢶㙈࡛┞பష⏕ࡌࡾࢰࣤࣂࢠ㈻ࡡࢪࢠ࣭ࣛࢼࣤࢡ
୷ᒜ₮ୌ 1ࠉᮿᑹୌ㑳 2ࠉ௿⸠ᖶᠺ 2 ࠉ໪ᮇົࡥࡆ 1  㸝1 ᮶ኬ㝌㎨⏍⛁࣬ᚺ⏍ᕝࠉ2 ⌦◂㸞㯔Ⳟ A. oryzae ࡢኬ㔖ࡡࢰࣤࣂࢠ㈻ࢅᇰᆀ୯࡞ฦἢࡌࡾ⬗ງࢅ᭯ࡊ࡙࠷ࡾࡆ࡛࠾ࡼࠉ᭯⏕ࢰࣤࣂࢠ㈻⏍⏐ࡡᐙ୹࡛ࡊ࡙⏕࠷ࡼࡿ࡙࠷ࡾࠊࡊ࠾ࡊࠉA. oryzae ࢅጙࡴ࡛ࡊࡒ⣊≟Ⳟ࡚ࡢฦἢ⤊㊪࡚ࢰࣤࣂࢠ㈻࠿ུࡄࡾဗ㈻⟮⌦࠽ࡻࡦ㐽ืᶭᵋ࡞ࡗ࠷࡙ࡡ◂✪ࡢ㝀ࡼࡿ࡙࠷ࡾࠊୌ⯙࡞ࠉᑚ⬂మ࡚ࡢࣆ࢚࣭ࣜࢸ࢔ࣤࢡ୘ධࡡࢰࣤࣂࢠ
㈻࠿ဗ㈻⟮⌦ᶭᵋ࡞ࡻࡽฦよࡈࡿࡾ࠿ࠉ᭩㎾ࡆࡡ㐛⛤࡚ࡡ N-⤎ྙᆵ⢶㙈ࡡᙲ๪࠿㓕ẍࡷິ∸࡚᪺ࡼ࠾࡞࡝ࡽ
ࡗࡗ࠵ࡾࠊᮇ◂✪࡚ࡢ⣊≟Ⳟ࡞࠽ࡄࡾ N-⤎ྙᆵ⢶㙈ࡡᶭ⬗よ᪺ࢅ┘Ⓩ࡛ࡊ࡙ࠉࡆࡡ⢶㙈࡛┞பష⏕ࡌࡾࢰࣤ
ࣂࢠ㈻ࡡࢪࢠ࣭ࣛࢼࣤࢡࢅ⾔ࡖࡒࠊ
A. oryzae ࡡⳞమ᢫ฝ∸࠾ࡼ㐪ᚨฦ㞫࡞ࡻࡽࠉᑚ⬂మࢨࣔ࣋ࣞࣤ BipA ࢅᣞᵾ࡞ᑚ⬂మࢅྱࡳ⭯⏤ฦࢅㄢ⿿
ࡊࡒࠊN-⤎ྙᆵ⢶㙈ࡡࡥ࡛ࡗ Glc1 Man9GlcNAc2 ࢅᅖᏽ໩ࡊࡒࣄ࣭ࢫ࡛࢕࣭ࣤ࢞ࣖ࣊ࢹࡊࡒ⤎ᯕࠉ⁈ฝ⏤ฦ࡞⣑ 70 kDa ࡡࢰࣤࣂࢠ㈻࠿≁␏Ⓩ࡞᳠ฝࡈࡿࡒࠊࡆࡿࢅࢹࣛࣈࢨࣤ࡞ࡻࡽᾐ໩ࡊࠉ⏍ࡋࡒ࣋ࣈࢲࢺ᩷∞ࢅ
LC/MS/MS ࡞ࡻࡽよᯊࡊࡒࠊᚋࡼࡿࡒ࣋ࣈࢲࢺࡡฦᏄ㔖࠽ࡻࡦහ㒂࢓࣐ࢿ㓗㒼าࢅࡵ࡛࡞ࠉA. oryzae ࢣࢿ࣑ࢸ࣭ࢰ࣭࣊ࢪࢅ᳠⣬ࡊࡒ࡛ࡆࢀࠉᑚ⬂మࢨ࡚ࣔ࣋ࣞࣤ࠵ࡾ࢜ࣜࢾ࢞ࢨࣤࡡࢸ࣭ࢰ࡛᭯ណ࡞ୌ⮬ࡊࡒࠊ࢜ࣜࢾ
࢞ࢨࣤ࠿ Glc1 Man9GlcNAc2 ࡛⤎ྙࡌࡾࡆ࡛ࡢࡆࡿࡱ࡚࡞ሒ࿈ࡈࡿ࡙࠷ࡾࡆ࡛࠾ࡼࠉᮇ◂✪ࡡᐁ㥺⣌࠿᭯ຝ࡚࠵ࡾࡆ࡛࠿☔ヾࡈࡿࡒࠊ⌟ᅹࠉN-⤎ྙᆵ⢶㙈࡛┞பష⏕ࡌࡾ௙ࡡࢰࣤࣂࢠ㈻ࡡࢪࢠ࣭ࣛࢼࣤࢡࢅ⾔ࡖ࡙࠷ࡾࠊ
Screening for proteins that associate with N-glycan from A. oryzae
Jun-ichi Maruyama1, Ichiro Matsuo2, Yukishige Ito2, Katsuhiko Kitamoto1(1Dept of Biotechnology, Univ of Tokyo, 2RIKEN)

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㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ ER-Golgi మ㛣ࡡ㍲㏞࡞㛭ࢂࡾ SNARE ࡡᑻᅹよᯊ⏛὾⥜Ꮔ㸡ಲ㒌ᑑᏳ, ḿ㊪ῗஒ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞࠘┘Ⓩ࠙A. oryzae ࡢ࢓࣐࣭ࣚࢭࠉࣈࣞࢷ࢓࣭ࢭ࡝࡜ࡡࢰࣤࣂࢠ㈻ࢅኬ㔖࡞ฦἢࡌࡾ⬗ງࢅᣚࡗ࠿ࠉ⣵⬂හᑚ⬂㍲㏞࡞㛭ࡌࡾฦᏄ࡚ࣝ࣊ࣜࡡ▩ぜࡢᑛ࡝࠷ࠊඁࡿࡒฦἢ⬗ງࢅ᭯ࡌࡾ A. oryzae ࡞࠽࠷࡙ฦἢ⤊㊪ࡡ◂✪ࡢᚺ⏕㟻࡚ࡵ㔔さ࡚࠵ࡽࠉࡆࡿࢅ⌦よࡌࡾࡒࡴ࡞ࡢࡐࡡ᭩ิࡡࢪࢷࢴࣈࡡᑚ⬂㍲㏞࡞㛭ࡌࡾ▩ぜࡡ⵫✒࠿ᙲ❟ࡗ࡛⩻࠻ࡒࠊࡐࡆ࡚ࠉᑚ⬂㍲㏞ࢅ௯௒ࡌࡾ SNARE ࡡ࠹ࡔࠉ≁࡞ ER-Golgi మ㛣഼࡚ࡂࡵࡡ࡞Ἰ┘ࡊࡒࠊ࠘᪁Ἢཀྵࡦ⤎ᯕ࠙S. cerevisiae ࡞࠽࠷࡙ ER-Golgi మ㛣഼࡚ࡂ࡛⩻࠻ࡼࡿࡾ SNARE ࡡࠉA. oryzae ࡞࠽ࡄࡾ࣓࣌
ࣞࢡࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ 7 ಴㸝Aobet1ࠉAobos1ࠉAosec22ࠉAoufe1ࠉAosec20ࠉAouse1ࠉAogos1㸞ࢅ༟㞫ࡊࡒࠊ྘㐿ఎᏄ࡞ࡗ࠷࡙ࠉMultiSite GatewayTM Cloning System ࢅ⏕࠷࡙ amyB ࣈ࣓࣭ࣞࢰ࣭ୖ࡚ egfp ࡛ࡡ⼝ྙ㐿ఎᏄࢅⓆ⌟ࡌࡾ࣊ࢠࢰ࣭㸝྘㐿ఎᏄࡡ 5̓ᮆ❻ഁ࡞ egfp ࢅ⼝ྙ㸞ࢅᵋ⠇ࡊࠉࡆࡿࡼࢅ⏕࠷࡙ᙟ㈻㌹ᥦࢅ⾔ࡖࡒࠊཱིᚋࡊࡒᰬࡡ㢟᚜㙶ふᐳࢅ⾔࠷ࠉࡆࡿࡼࡡ SNARE ࡡᑻᅹ㒂నࢅ࢛ࣜ࢝ࢾࣚ᯹Ⰵモ࡛ⷾࡡභ᯹Ⰵ࡞ࡻࡖ࡙ྜྷ
ᏽࡊࡒ࡛ࡆࢀࠉA. oryzae ࡞࠽࠷࡙ࡵ S. cerevisiae ࡛ྜྷᵕࡡᑻᅹࢅ♟ࡌࡆ࡛࠿♟ြࡈࡿࡒࠊ௧୕ࡡ⤎ᯕ࡞ࡻࡽࠉ
௑ᚃࡆࡿࡼࡡ SNARE ࢅ྘࢛ࣜ࢝ࢾࣚࡡ࣏࣭࣭࡛࢜ࡊ࡙౐⏕ࡌࡾ࡝࡜ࠉA. oryzae ࡞࠽ࡄࡾ ER-Golgi మ㛣ࡡ㍲㏞よᯊ࡞㐲⏕ࡌࡾࡆ࡛࠿࡚ࡀࡾ࡛⩻࠻ࡼࡿࡾࠊ
Localization analysis of SNARE proteins that function between ER and Golgi compartments in A. oryzae
Ayako TAURA, Masahiro KURATSU, Jun-ya SHOJI, Manabu ARIOKA, Katsuhiko KITAMOTO(Dept. of Biotech., Univ. of Tokyo)

P-46
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ post-Golgi ㍲㏞࡞㛭ࢂࡾ SNARE ࡡᑻᅹよᯊಲ㒌ᑑᏳ㸡ḿ㊪ῗஒ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞࠘┘Ⓩ࠙ᠻࠍࡢࡆࡿࡱ࡚ A. oryzae ࡞࠽࠷࡙ ERࠉGolgi మࠉᾦ⬂࡞ᑻᅹࡌࡾࢰࣤࣂࢠ㈻࡛ EGFP ࡛ࡡ⼝ྙࢰࣤ
ࣂࢠ㈻ࢅⓆ⌟ࡈࡎࡾࡆ࡛࡞ࡻࡽࠉࡐࡿࡼࡡ࢛ࣜ࢝ࢾࣚࡡྊっ໩ࢅ⾔ࡖ࡙ࡀࡒࠊࡊ࠾ࡊࠉGolgi మ௧㜾ࡡ࢛ࣜ࢝ࢾࣚࡡ✭㛣Ⓩฦᕱ࡞㛭ࡌࡾ⣵⬂⏍⌦ᏕⓏ࡝▩ぜࡢኣࡂ࡝࠷ࠊࡐࡆ࡚ࠉ྘࢛ࣜ࢝ࢾࣚ࡞ᑻᅹࡊࠉ㍲㏞ᑚ⬂࡛ᵾ
Ⓩ࢛ࣜ࢝ࢾࣚ⭯ࡡ⤎ྙཀྵࡦ⼝ྙࡡ≁␏ᛮࢅᢰ࠹ SNARE ࢰࣤࣂࢠ㈻࡞Ἰ┘ࡊࡒࠊᮇ◂✪࡚ࡢࠉS. cerevisiae ࡡ
SNARE ࢰࣤࣂࢠ㈻ࡡ㒼าࢅᇱ࡞ A. oryzae ࡡࢣࢿ࣑᝗ሒ࠾ࡼ 21 ಴ࡡ SNARE ࢅ᥆ᏽࡊࠉࡐࡡ୯࠾ࡼ post-Golgi
ࡡ㍲㏞࡞㛭ࢂࡾ࡛஢᝷ࡈࡿࡾࡵࡡ࡞ࡗ࠷࡙ᑻᅹよᯊࢅ⾔ࡖࡒࠊ࠘᪁Ἢཀྵࡦ⤎ᯕ࠙ ᥆ᏽ SNARE ࢰࣤࣂࢠ㈻ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ 12 ಴㸝Aosft1ࠉAoykt6ࠉAovti1ࠉAotlg1ࠉAotlg2ࠉ
Aosyn8ࠉAonyv1ࠉAovam7ࠉAosso1ࠉAosso2ࠉAosec9ࠉAosnc1㸞࡞ࡗ࠷࡙ࠉegfp ࡛ࡡ⼝ྙ㐿ఎᏄࢅⓆ⌟ࡌࡾ࣊ࢠࢰ࣭ࢅ⏛὾ࡼ࡛ྜྷᵕࡡ᪁Ἢ࡚ᵋ⠇ࡊࡒࠊࡆࡿࡼࢅ⏕࠷࡙ᙟ㈻㌹ᥦࢅ⾔࠷ࠉ྘ SNARE ࢰࣤࣂࢠ㈻ࡡྊっ໩ࢅ⾔ࡖࡒࠊEGFP ⺧කࡡふᐳ࡛⣵⬂᯹Ⰵࡡ⤎ᯕ࠾ࡼࠉ12 ಴ࡡ᥆ᏽ SNARE ࢰࣤࣂࢠ㈻ࡢ S. cerevisiae ࡛࡮࡯ྜྷᵕࡡ࢛ࣜ࢝ࢾࣚ࡞ᑻᅹࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊࡱࡒ྘࢛ࣜ࢝ࢾࣚ࡞ࡗ࠷࡙ࠉGolgi మࡢⳞ⣊඙❻࡞ኣࡂᏋᅹࡊࠉ
late endosome/PVC ࡢᾦ⬂࡞㎾᥃ࡊ࡙ᏋᅹࡌࡾࠉࡐࡿࡑࡿⅤ≟ࡡᵋ㏸మ࡛ࡊ࡙⣵⬂හ࡞ฦᕱࡌࡾࡆ࡛࠿♟ြࡈ
ࡿࡒࠊ

Localization analysis of SNARE proteins that function in post-Golgi transport in A. oryzae
Masahiro KURATSU, Jun-ya SHOJI, Manabu ARIOKA, Katsuhiko KITAMOTO(Dept. of Biotech., Univ. of Tokyo)

73

P-47
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ᥃ྙࣆ࣓࢘ࣞࣤ๑㥉మ㐿ఎᏄ AoppgA ࡡᶭ⬗よᯊ
ᒜᮇ୏℡ࠉJUVVADI Praveen Raoࠉ୷ᒜ₮ୌࠉ໪ᮇົࡥࡆ (᮶ኬ㝌㎨⏍⛁࣬ᚺ⏍ᕝ)
࠘┘Ⓩ࠙㯔Ⳟ A. oryzae ࡢࡆࡿࡱ࡚࡞᭯ᛮୠ௥࠿☔ヾࡈࡿ࡙࠷࡝࠷ࡵࡡࡡࠉࢣࢿ࣑୯࡞ࡢኣࡂࡡ᥃ྙ㛭㏻㐿ఎᏄࡡ┞ྜྷ㐿ఎᏄ࠿Ꮛᅹࡌࡾࠊࡐࡡୌࡗ࡚࠵ࡾȘ-ࣆ࣓࢘ࣞࣤ๑㥉మ㐿ఎᏄ AoppgA ࡢࠉ᭯ᛮୠ௥ࡡ☔ヾࡈࡿ࡙࠷ࡾ⣊≟Ⳟ࡞࠽࠷࡙ࠉ᥃ྙ┞ᡥࡡヾㆉࡷུ⢥ᚃࡡᰶ⼝ྙ࡞࠽࠷࡙㔔さ࡝഼ࡀࢅࡊ࡙࠷ࡾ࡛⩻࠻ࡼࡿ࡙࠷ࡾࠊ
ࡊ࠾ࡊࠉ᭯ᛮୠ௥ࡡ▩ࡼࡿ࡙࠷࡝࠷⣊≟Ⳟ࡞࠽ࡄࡾᶭ⬗ࡢᮅ▩࡚࠵ࡾࠊࡐࡡࡒࡴ୘ᏰධⳞ㢦࡛⩻࠻ࡼࡿ࡙࠷
ࡾ㯔Ⳟ࡞࠽࠷࡙ࠉ᥃ྙࣆ࣓࢘ࣞࣤࡡ᩺ࡒ࡝ᶭ⬗ࡡよ᪺ࢅ┘ᣞࡊ࡙よᯊࢅ⾔ࡖࡒࠊ࠘᪁Ἢཀྵࡦ⤎ᯕ࠙Ș-ࣆ࣓࢘ࣞࣤࡡᶭ⬗ࢅ᥀⣬ࡌࡾࡒࡴࠉȘ-ࣆུ࣓࢘ࣞࣤᐖమࡡ┞ྜྷ㐿ఎᏄ AogprA ࢅ༟㞫ࡊ
ࡒࠊAoGprA ࡢ 369 ࢓࣐ࢿ㓗ࡻࡽ࡝ࡽࠉ㸬ᅂ⭯㈇㏳㡷ᇡࢅࡵࡗ୔㔖మ G ࢰࣤࣂࢠ㈻භᙲᆵུᐖమ࡚࠵ࡾࠊ௙
ࡡ Aspergillus ᒌࡡȘ-ࣆུ࣓࢘ࣞࣤᐖమ㐿ఎᏄ࡛Ẓ㍉ࡌࡾ࡛ࢰࣤࣂࢠ㈻࡚ࣝ࣊ࣜ⣑ 50㸚ࡡ┞ྜྷᛮࢅ♟ࡊࡒࠊ
RT-PCR よᯊ࡞ࡻࡽࠉAoppgAࠉAogprA 㐿ఎᏄࡢ࡛ࡵ࡞Ⓠ⌟ࡊ࡙࠷ࡾࡆ࡛ࠉࡱࡒ⅛⣪″ᯜῤ࠽ࡻࡦ࢓᮪ࣜ࢜ࣛ
௲ୖ࡚Ⓠ⌟ࡡ୕᪴ࡌࡾࡆ࡛࠿ヾࡴࡼࡿࡒࠊAoppgAࠉAogprA 㐿ఎᏄ㧏Ⓠ⌟ᰬࠉ࠽ࡻࡦ◒ቪᰬࢅష⿿ࡊࠉ⏍⫩㏷
ᗐࠉฦ⏍Ꮔᩐࠉᙟឺ࡞࠽ࡄࡾ⾪⌟ᆵࡡよᯊࢅ⾔ࡖ࡙࠷ࡾࠊ

Functional analysis of alpha-pheromone precursor AoppgA in Aspergiilus oryzae
Nanase Yamamoto, JUVVADI Praveen Rao, Jun-ichi Maruyama, Katsuhiko Kitamoto(Dept. of Biotechnol., Univ. of Tokyo)

P-48ࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪ࡞ࡻࡾ㯔Ⳟ A. oryzae ࢅ⏕࠷ࡒࣃࢹࣛࢯࢲ࣭࣑⏍⏐㔖ࡡቌຊ
ῳ㎰ὀ♰㸡㔘㗋᮵㸡୷ᒜ₮ୌ㸡Praveen Rao Juvvadi㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞࠘┘Ⓩ࠙ࢰࣤࣂࢠ㈻㧏ฦἢ⬗ࢅ᭯ࡌࡾ㯔Ⳟ A. oryzae ࡢࠉ␏⛸ࢰࣤࣂࢠ㈻⏍⏐ࡡᐙ୹࡛ࡊ࡙Ἰ┘ࡈࡿ࡙࠷ࡾࠊ
ࡊ࠾ࡊࡆࡿࡱ࡚࡞ࠉA. oryzae ࡞࠽࠷࡙ࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪ࡞ࡻࡽ␏⛸ࢰࣤࣂࢠ㈻ࡡ⏍⏐㔖࠿ቌຊࡊࡒ࡛
࠷࠹ሒ࿈ࡢ࡝࠷ࠊⁿ⩽ࡼࡢࠉࣃࢹࣛࢯࢲ࣭࣑⏍⏐ࢅȘ-࢓࣐࣭ࣚࢭ㸝AmyB㸞ࢅ࢞ࣔࣛ࢓࣭࡛ࡊ࡙⾔࠷ࠉ5DPY ᾦమᇰᆀ㸝pH 8.0㸞࡞࠽࠷࡙᭩ኬ 15.6 mg/L ࡡ⏍⏐㔖ࢅᚋ࡙࠷ࡾ 1)ࠊ௑ᅂࡢࠉ㸪⛸ࡡࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ㸝pepA, pepE, alpA, tppA, palB㸞ࡡୌ㔔࠽ࡻࡦ஦㔔ࡡ◒ቪᰬ࡞ࡗ࠷࡙ࠉࣃࢹࣛࢯࢲ࣭࣑⏍⏐࡫ࡡຝᯕࢅ᳠ゞࡊࡒࠊ࠘᪁Ἢ࡛⤎ᯕ࠙㸪⛸㢦ࡡࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ◒ቪᰬࡡࣃࢹࣛࢯࢲ࣭࣑⏍⏐ᛮࢅ 5DPY ᾦమᇰᆀ㸝pH 8.0㸞࡞࠽࠷࡙Ẓ㍉ࡊࡒࠊࡐࡡ⤎ᯕࠉǼtppA, ǼpalB, ǼpepE, ǼalpA, ǼpepA ࡡ㡨࡞㐿ఎᏄ◒ቪࡡຝᯕ࠿ヾࡴࡼࡿࠉ࡝࠾࡚ࡵǼtppA ᰬࡡ⏍⏐㔖ࡢ 21.2 mg/L ࢅ♟ࡊࡒࠊࡈࡼ࡞ࠉǼpalB ᰬ࠽ࡻࡦǼpepE ᰬ࡞ᑊࡊ࡙ argB ࢅ㐽ᢝ࣏࣭࣭࡛࢜ࡊ࡙ tppA 㐿ఎᏄࢅ◒ቪࡌࡾࡆ࡛࡞ࡻࡽࠉࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄ஦㔔◒ቪᰬࢅష⿿ࡊࡒࠊࡆࡿࡼࡡᰬ
ࢅ⏕࠷࡙ࣃࢹࣛࢯࢲ࣭࣑⏍⏐ࢅ⾔ࡖࡒ࡛ࡆࢀࠉǼtppAǼpepE ஦㔔◒ቪᰬ࡞࠽࠷࡙᭩ࡵ㧏࠷⏍⏐㔖㸝25.4 mg/L㸞࠿ᚋࡼࡿࡒࠊ௧୕ࡡ⤎ᯕ࠾ࡼࠉA. oryzae ࢅ⏕࠷ࡒࣃࢹࣛࢯࢲ࣭࣑⏍⏐࡞࠽࠷࡙ࠉࣈࣞࢷ࢓࣭ࢭ㐿ఎᏄࡡ஦㔔◒ቪࢅ⾔࠹ࡆ࡛࡞ࡻࡽ⣑ 1.6 ಶࡡ⏍⏐㔖ࡡቌຊ࡞ᠺຉࡊࡒࠊ
1) 㔘ࡼࠉ➠㸩ᅂ⣊≟ⳞฦᏄ⏍∸Ꮥࢤࣤࣆ࢒ࣝࣤࢪさ᪠㞗 p. 39

Higher-level production of human lysozyme by protease gene disruption in A. oryzae
Taisuke Watanabe, Feng Jie Jin, Jun-ichi Maruyama, Praveen Rao Juvvadi, Manabu Arioka, Katsuhiko Kitamoto㸝Dept.
of Biotech., Univ. of Tokyo㸞

74

P-49
㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ 2 ࡗࡡฦἢᆵ࣌ࢪ࣌ࣛࣂ࣭ࢭ A2 ࡡᢘమࢅ⏕࠷ࡒᑻᅹよᯊ࡛
RT-PCR ࡞ࡻࡾⓆ⌟よᯊ
୯℀ᬓ஄㸡໪ᮇົࡥࡆ㸡᭯ᒱ Ꮥ 㸝᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ㸞࠘┘Ⓩ࠙ฦἢᆵ࣌ࢪ࣌ࣛࣂ࣭ࢭ A2 (sPLA2) ࡢࢡࣛࢬࣞࣛࣤ⬙㈻ࡡ 2 నࡡ࢙ࢪࢷࣜ⤎ྙࢅຊỀฦよࡊࠉ⬙⫣㓗࡛ࣛࢯࣛࣤ⬙㈻ࢅ㐗㞫ࡌࡾ㓕⣪࡚࠵ࡾࠊဳ஘㢦࡞࠽࠷࡙ࡢ⣑ 10 ಴ࡡ sPLA2 㐿ఎᏄ࠿ぜฝࡈࡿᶭ⬗よᯊ࠿⾔ࢂࡿ࡙࠷ࡾ࠿ࠉ᚜⏍∸⏜ᮮࡡ sPLA2 ࡡ⏍⌦Ⓩ࡝ᙲ๪ࡢṜ࡜᪺ࡼ࠾࡞ࡈࡿ࡙࠷࡝࠷ࠊᮇ◂✪࡚ࡢ㯔Ⳟࡡᣚࡗ 2 ࡗ
ࡡ sPLA2㸝SpaA, SpaB㸞ࡡよᯊࢅ⾔ࡖࡒࠊ࠘᪁Ἢ࠙ࡆࡿࡱ࡚࡞㧏Ⓠ⌟ᰬ࡞࠽ࡄࡾ PLA2 Ὡᛮࡡฦᕱࢅㄢ࡬ࡒ⤎ᯕ࠾ࡼࠉSpaA ࠿୹࡞ᇰᆀ୯࡞ฦἢࡈࡿࡾ
ࡡ࡞ᑊࡊࠉSpaB ࡢ஢᝷ࢨࢡࢻࣜ㒼าࢅᣚࡗ࡞ࡵ㛭ࢂࡼࡍฦἢࡈࡿࡍⳞమ⏤ฦ࡞ᑻᅹࡌࡾࡆ࡛࠿♟ြࡈࡿ࡙࠷
ࡾࠊ௑ᅂࠉ⤄ᥦ࠻ SpaA ࠽ࡻࡦ SpaB ࢰࣤࣂࢠ㈻ࢅᢘཋ࡛ࡊ࡙྘ࠍࡡࢰࣤࣂࢠ㈻࡞≁␏Ⓩ࡝࣎ࣛࢠ࣭ࣞࢻࣜᢘ
మࢅష⿿ࡊࠉࡻࡽレ⣵࡝᳠ゞࢅ⾔ࡖࡒࠊ࢙ࢗࢪࢰࣤࣇࣞࢴࢹ࡞ࡻࡽ㧏Ⓠ⌟ᰬࡡᇰ㣬୕ΰ⏤ฦ࡛Ⳟమ⏤ฦࢅよᯊࡊࡒ࡛ࡆࢀࠉ୕㏑ࡡ⤎ᯕ࡛ୌ⮬ࡊ࡙ SpaA ࠿ᇰ㣬୕ΰ࡞ࠉSpaB ࠿Ⳟమ⏤ฦ࡞Ꮛᅹࡌࡾࡆ࡛࠿ࢂ࠾ࡖࡒࠊࡆ
ࡡࡆ࡛࠾ࡼ୦ࢰࣤࣂࢠ㈻࠿␏࡝ࡾᑻᅹࢅ♟ࡌࡆ࡛࠿ッ᪺ࡈࡿࡒࠊ⌟ᅹࠉ㛣᥃⺧කᢘమἪ࡞ࡻࡾふᐳࢅ⾔࡝࠷ࠉ୦ࢰࣤࣂࢠ㈻ࠉ≁࡞ SpaB ࡡ⣵⬂හ࡞࠽ࡄࡾᑻᅹࢅࡈࡼ࡞ㄢ࡬࡙࠷ࡾࠊࡱࡒࠉ㯔Ⳟࢅᵕࠍ࡝᮪௲࡚ᇰ㣬ࡊࠉ྘
㐿ఎᏄࡡⓆ⌟ࢅ RT-PCR Ἢ࡞ࡻࡖ࡙᳠ฝࡊࡒࠊࡐࡡ⤎ᯕࠉspaA ࡢࢿࢧࣤよᯊ࡞ࡻࡖ࡙ࡌ࡚࡞᪺ࡼ࠾࡞ࡈࡿ࡙
࠷ࡒ⅛⣪″ᯜῤ᮪௲࡞࠽ࡄࡾⓆ⌟࡞ຊ࠻࡙ࠉ✽⣪″ࡡᯜῤ࡛࢓᮪ࣜ࢜ࣛ௲࡞ࡻࡖ࡙ࠉࡱࡒ spaB ࡢ㸩Υ࡛࠷࠹఩Ὼ᮪௲࡞࠽࠷࡙Ⓠ⌟ࡌࡾࡆ࡛࠿ฦ࠾ࡖࡒࠊࡆࡿࡼࡡ᝗ሒࢅඔ࡞ࡆࡿࡱ࡚࡞ཱིᚋࡊ࡙࠷ࡾ spaAࠉspaB ༟≺ࠉ࠽ࡻࡦ spaA/spaB ஦㔔◒ቪᰬࡡ⾪⌟ᆵࢅ᥀⣬ࡌࡾࡆ࡛࡚ࠉࡐࡿࡑࡿࡡ㓕⣪ࡡ⏍⌦Ⓩ࡝ᶭ⬗ࢅྜྷᏽ࡚ࡀࡾ࡛᭿
ᙽࡊ࡙࠷ࡾࠊ
Molecular characterization of two secretory phospholipases A2 in Aspergillus oryzae.
Tomoyuki Nakahama, katsuhiko Kitamoto, and Manabu Arioka (Dept. of Biotechnol., Univ. of Tokyo㸞

P-50
㯔Ⳟ A. oryzae ࢅ⏕࠷ࡒࢹࣚࣆࢡ࣬ࣂࣆࣝࢠࢲࣤࡡ⏍⏐➪ᰡᚷಕ, 㤷ᶣ⏜౅, ୷ᒜ₮ୌ, ᭯ᒱᏕ, ໪ᮇົࡥࡆ (᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ)ⁿ⩽ࡼࡢࠉࢹࣚࣆࢡ⏜ᮮࡡࣝࢠࢲ࡚ࣤ࠵ࡾࣂࣆࣝࢠࢲࣤࢅࠉN ᮆ࡞࢞ࣔࣛ࢓࣭࡛ࡊ࡙Ș-࢓࣐࣭ࣚࢭ㸝AmyB㸞ࠉC ᮆ࡞᳠ฝ࠽ࡻࡦ⢥⿿ࡡࢰࢡ࡛ࡊ࡙ HA-His6 ࡛⼝ྙࡊ࡙⏍⏐ࢅ⾔ࡖࡒࠊࡱࡒࡆࡡ㝷࡞ࠉKex2 ᵕࣈࣞࢷ࢓࣭ࢭ
ヾㆉ㒼าࢅ AmyB ࡛ࡡ㏻⤎㒂న࡞ᑙථࡊࡒࠊࡆࡡࡻ࠹࡞ࡊཱི࡙ᚋࡊࡒࣂࣆࣝࢠࢲࣤ⏍⏐ᰬࡡᇰᆀ୕ΰ࡞ࡗ࠷
࡙ HA ᢘమࢅ⏕࠷࢙ࢗࢪࢰࣤよᯊࢅ⾔ࡖࡒ࡛ࡆࢀࠉ┘Ⓩࡡ⣑ 16 kDa ࡡ⏍⏐∸࠿☔ヾࡈࡿࡒ࠿ࠉAmyB ࡛ࡡ⼝ྙࢰࣤࣂࢠ㈻ࡵ᳠ฝࡈࡿࡒࠊࡐࡆ࡚ࠉ௑ᅂࡢࠉࣂࣆࣝࢠࢲ࡛ࣤ Kex2 ᵕࣈࣞࢷ࢓࣭ࢭヾㆉ㒼าࡡ㛣࡞ࠉࢡࣛࢨ
ࣤࢅ㸨ṟᇱ᣼ථࡊࡒࢤࣤࢪࢹࣚࢠࢹࢅషᠺࡊࠉ㯔Ⳟ࡞ᑙථࡊࡒࠊᚋࡼࡿࡒᰬࡡᇰᆀ୕ΰ࡞ࡢࠉ⼝ྙࢰࣤࣂࢠ㈻ࡢぜࡼࡿࡍࠉࣂࣆࣝࢠࢲࣤࡡࡲ࠿☔ヾࡈࡿࡒࠊࡈࡼ࡞ࠉࢼࢴࢢ࣑ࣜ࢜ࣚࢅ⏕࠷ His6 ࢰࢡࢅ᭯ࡌࡾࢰࣤࣂࢠ
㈻ࢅ⢥⿿ࡊࡒ࡛ࡆࢀࠉ⢥⿿⏤ฦ࡞ࡢ┘Ⓩࡡࢦ࢕ࢫࡡࡵࡡࡡ௙࡞ HA ᢘమ࡚ࡢヾㆉࡈࡿ࡝࠷ࡷࡷᑚࡈࡴࡡ⏍⏐∸ࡵ᳠ฝࡈࡿࡒࠊN ᮆࢨ࣭ࢠ࢙ࣤࢪࡡ⤎ᯕࠉ୦ࢰࣤࣂࢠ㈻࡛ࡵࣂࣆࣝࢠࢲࣤࡡ୕Ὦ࡞᣼ථࡊࡒ㸨ࡗࡡࢡࣛࢨ
ࣤ࠾ࡼጙࡱࡾ࢓࣐ࢿ㓗㒼าࢅࡵࡖ࡙࠷ࡒࡆ࡛࠾ࡼࠉᑚࡈ࡝ࢰࣤࣂࢠ㈻ࡢࢰࢡࢅྱࡳ C ᮆ❻࠿Ḗ኶ࡊࡒࡵࡡ࡛
⩻࠻ࡼࡿࡒࠊࡆࡿࡼࡡ⤎ᯕࡢࠉHA-His6 ࢰࢡ㒼าࢅヾㆉࡌࡾ࣋ࣈࢲࢱ࣭ࢭࡡᏋᅹ࠿஢᝷ࡈࡿࡒࡡ࡚ࠉC ᮆ❻ࡡฦよࢅ㜭ࡃࡒࡴ࡞ࢰࢡࡡ㒼าࢅᨭንࡊࡒࢤࣤࢪࢹࣚࢠࢹࢅషᠺࡊࠉࣂࣆࣝࢠࢲࣤࡡ⏍⏐ࢅモࡲ࡙࠷ࡾࠊ

Production of Takifugu rubripes puflectin by A. oryzae.
Shiho Sasaguri, Yuka Mabashi, Jun-ichi Maruyama, Manabu Arioka, Katsuhiko Kitamoto(Dept. of Biotechnology, Univ. of Tokyo)

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㯔Ⳟ A. oryzae ࡞࠽ࡄࡾ Aovps24 (class E vps)㐿ఎᏄࡡ༟㞫࡛よᯊ㎦ᕦᬏ⣎㸡ⳝ㛣㝧ᚷ㸡᭯ᒱᏕ㸡໪ᮇົࡥࡆ (᮶ኬ㝌㹺㎨⏍⛁㹺ᚺ⏍ᕝ)<┘Ⓩ>VPS (vacuolar protein sorting)㐿ఎᏄࡢฝⰾ㓕ẍ S. cerevisiae ࡞࠽࠷࡙ᾦ⬂㓕⣪ carboxypeptidase Y (CPY)ࢅ⣵⬂አ࡞ฦἢࡌࡾን␏࡞㛭ࡌࡾ㐿ఎᏄ࡚࠵ࡽࠉࢰࣤࣂࢠ㈻㍲㏞ᶭᵋࡷᾦ⬂ᶭ⬗࡝࡜࡞㛭୙ࡌࡾࡆ࡛࠿▩ࡼࡿ࡙࠷ࡾࠊࡱࡒ㎾ᖳࠉᚃ᭿࢙ࣤࢺࢮ࣭࣑࠾ࡼᾦ⬂࡫ࡡ㍲㏞ࢅྒྷࡾ multivesicular body (MVB)ࡡᙟᠺ࡞㛭ࢂࡾࢰ
ࣤࣂࢠ㈻࠿ࠉ࢓ࣜ࢜ࣛ pH ᚺ➽ࢅโᚒࡌࡾ Rim101 ⤊㊪࡞ࡵ㛭୙ࡌࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊᠻ࠿ᅗ࡚᪝ᮇ㒿ࡷ࿝ძࡡ㔂㏸࡞⏕࠷ࡼࡿ࡙࠷ࡾ㯔Ⳟ࡞࠽࠷࡙ࠉࢰࣤࣂࢠ㈻ࡡ㍲㏞⤊㊪ࡷᾦ⬂ᶭ⬗ࠉpH ᚺ➽ࡡよ᪺ࡢ㔔さ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡾࠊࡐࡆ࡚ࠉMVB ᙟᠺ࡞㛭ࢂࡾ class E vps 㐿ఎᏄࡡ୯ࡡ 1 ࡗ࡚࠵ࡾ Aovps24 㐿ఎᏄࡡᶭ⬗よᯊࢅ⾔ࡖࡒࠊ<᪁Ἢཀྵࡦ⤎ᯕ>A. oryzae ࡡࢣࢿ࣑ࢸ࣭ࢰ࣭࣊ࢪࢅᇱ࡞ VPS24 ࡛┞ྜྷᛮࡡ㧏࠷㐿ఎᏄ㸝Aovps24㸞ࢅ༟㞫ࡊࡒࠊ
Aovps24 ࡢ 1 ࡗࡡ࢕ࣤࢹࣞࣤࢅྱࡲࠉ228 ಴ࡡ࢓࣐ࢿ㓗࠾ࡼ࡝ࡾࢰࣤࣂࢠ㈻ࢅࢤ࣭ࢺࡌࡾ࡛᥆ᏽࡈࡿࡒࠊ
AoVps24 ࡢ S. cerevisiaeࠉS. pombe ࡡ Vps24 ࡛ࡐࡿࡑࡿ 34.1%ࠉ30.7%ࡡ࢓࣐ࢿ㓗┞ྜྷᛮࢅ♟ࡊࡒࠊAoVps24ࡡᶭ⬗ࢅㄢ࡬ࡾࡒࡴࠉྜྷ㐿ఎᏄࡡ◒ቪᰬࢅషᠺࡊࡒࠊᚋࡼࡿࡒ◒ቪᰬࡢⴥࡊ࠷⏍⫩㜴ᐐࢅ♟ࡊࠉⳞ⣊඙❻ࡡኣฦᒪࡷ⭶₮࡛࠷ࡖࡒᙟឺ␏ᖏࡵぜࡼࡿࡒࠊࡱࡒฦ⏍Ꮔᙟᠺࡵぜࡼࡿ࡝࠾ࡖࡒࠊࡆࡿࡼࡡࡆ࡛࠾ࡼࠉAoVps24
࠿ኣ㟻Ⓩ࡝ᶭ⬗ࢅᣚࡔࠉ⣊≟Ⳟ࡞࠽ࡄࡾᾦ⬂㍲㏞ࢰࣤࣂࢠ㈻࠿ᙟឺᙟᠺࡷฦ໩࡞ࡵ㛭୙ࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ⌟ᅹ A. oryzae ࡞࠽ࡄࡾ MVB ࡞㛭ࢂࡾࢰࣤࣂࢠ㈻࡞ࡗ࠷࡙ࡡࡈࡼ࡝ࡾ▩ぜࢅịࡴࠉAovps2ࠉAovps4 ࡡ◒ቪᰬࢅషᠺ୯࡚࠵ࡾࠊࡈࡼ࡞ AoVps24 ࡛ EGFP ࡡ⼝ྙࢰࣤࣂࢠ㈻ࡡⓆ⌟࡞ࡻࡽ┞⿭ᐁ㥺ཀྵࡦ⣵⬂හᑻᅹよᯊࢅ⾔ࡖ࡙࠷ࡾࠊࡱࡒ pH ᚺ➽࡞㛭ࡊ࡙ࡡ AoVps24 ࡡᶭ⬗ࢅよᯊ୯࡚࠵ࡾࠊ
Cloning and functional analysis of Aovps24 (class E vps) gene from A. oryzae
Akinori Tatsumi, Takashi Kikuma, Manabu Arioka, Katsuhiko Kitamoto(Dept. of Biotechnology, Univ. of Tokyo)

P-52
㯔Ⳟ A. oryzae ࡡ III ᆵ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕 ⣪㐿ఎᏄࡡⓆ⌟࡛ᶭ⬗よᯊເࠎᗛ௥ 1㸡⸠஬ ໌ 2㸡Praveen Rao Juvvadi1㸡ᾇ⩹ሪ㇇ 2㸡໪ᮇົࡥࡆ 1
㸝1 ᮶ኬ㝌࣬㎨⏍⛁࣬ᚺ⏍ᕝ, 2 ᮶ኬ㝌࣬ⷾ㸞࠘┘Ⓩ࠙III ᆵ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㸝PKS㸞࡛ࡊ࡙ࡢࠉ᳔∸ࡡ࢜ࣜࢤࣤྙᠺ㓕⣪ࡷࢪࢲࣜ࣊ࣤྙᠺ㓕⣪࠿▩ࡼࡿ࡙࠷ࡒ࠿ࠉ㎾ᖳࠉࣁࢠࢷࣛ࢓࠾ࡼࡵⰶ㤮᪐ྙᠺ㓕⣪࠿ぜฝࡈࡿࠉࡐࡡり፳ࡌࡾཬᚺࡡኣᵕᛮ࠾ࡼࠉ⌟ᅹࠉ㟸ᖏ࡞Ἰ┘ࡈࡿ࡙࠷ࡾࠊᠻࠍࡢࡆࡿࡱ࡚࡞ࠉ㯔Ⳟ A. oryzae ࡡࢣࢿ࣑୯࡞⣊≟Ⳟࡡ III ᆵ PKS 㐿ఎᏄ࡛ࡊ࡙ิ
ࡴ࡙ࡡౚ࡛࡝ࡾ 4 ࡗࡡ㐿ఎᏄ㸝csyAࠉcsyBࠉcsyC ࠽ࡻࡦ csyD㸞ࢅぜฝࡊࠉcsyA, B, D ࡡ 3 㐿ఎᏄ࠿ A. oryzae࡚Ⓠ⌟ࡊ࡙࠷ࡾࡆ࡛ࢅ☔ヾࡊ࡙࠷ࡾࠊᮇ◂✪࡚ࡢࠉࡆࡿࡼ㐿ఎᏄࡡᶭ⬗ࢅよ᪺ࡌࡾࡒࡴࠉ㯔Ⳟ A. oryzae ࢅᐙ୹࡛ࡊࡒ྘㐿ఎᏄ㧏Ⓠ⌟ᰬ࠽ࡻࡦ◒ቪᰬࡡష⿿ࢅ⾔࠷ࠉ⏍⏐໩ྙ∸ࡡよᯊࠉ࡝ࡼࡦ࡞ኬ⭘Ⳟ࡞ࡻࡾ⤄ᥦ࠻ࢰ
ࣤࣂࢠࡡⓆ⌟࡛ᶭ⬗よᯊࢅモࡲࡒࠊ࠘᪁Ἢ࠽ࡻࡦ⤎ᯕ࠙csyAࠉcsyBࠉcsyC ࠽ࡻࡦ csyD 㐿ఎᏄ࡞ࡗ࠷࡙ࠉ㯔Ⳟ A. oryzae ࢅᐙ୹࡛ࡊࡒ㧏Ⓠ⌟ᰬ࠽ࡻࡦ◒ቪᰬࡡష⿿ࢅ⾔ࡖࡒࠊcsyA 㐿ఎᏄ㧏Ⓠ⌟ᰬ࡞࠽࠷࡙ࠉࢤࣤࢹ࣭ࣞࣜࡡᙟ㈻㌹ᥦమ࡞ࡢぜࡼࡿ࡝࠷Ⰵ⣪ࡡ⏍⏐࠿ヾࡴࡼࡿࡒࠊᇰ㣬୕ΰࢅ LC/MS ࡚ฦᯊࡊࡒ⤎ᯕࠉᇰ㣬᪝ᩐ࡞ࡻࡖ࡙␏࡝ࡾᴗᛮ࣬᥆ᏽฦᏄ㔖ࡡ໩ྙ∸ࡡࣅ࣭ࢠ࠿ヾࡴࡼࡿࡾࡆ࡛࠾ࡼࠉCsyA ࡡ⏍ྙᠺ໩ྙ∸ࡢㄇᑙమ໩ࡈࡿ࡙࠷ࡾࡵࡡ࡛஢᝷ࡈࡿࡒࠊࡱࡒࠉ
RACE Ἢ࠽ࡻࡦ㐿ఎᏄᙁโⓆ⌟ᰬࢅ⏕࠷࡙ cDNA ࢅཱིᚋࡊࠉኬ⭘Ⳟࢅᐙ୹࡛ࡊ࡙ CsyAࠉCsyB ࠽ࡻࡦ CsyC྘⤄ᥦ࠻ࢰࣤࣂࢠࢅㄢ⿿ࡊࡒࠊ⌟ᅹࠉ⤄ᥦ࠻ࢰࣤࣂࢠࢅ⏕࠷࡙ in vitro ࡚ࡡὩᛮ☔ヾࢅ㐅ࡴ࡙࠷ࡾࠊ
Expression and functional analysis of type III polyketide synthase genes from Aspergillus oryzae
Yasuyo Seshime1, Isao Fujii2, Praveen Rao Juvvadi1, Yutaka Ebizuka2, Katsuhiko Kitamoto1(1Dept. of Biotechnology, Univ. of Tokyo, 2Grad. School of Pharm. Sciences, Univ. of Tokyo)

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Penicillium luteo-aurantium ࡡ᩺ぞ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㐿ఎᏄࡡࢠ࣭ࣞࢼࣤࢡ ࡛ᶭ⬗よᯊఫ㔕⿩ᇱࠉ▴஬⤦⨶ࠉ⸠஬ ໌ࠉᾇ⩹ሪ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞⣵⬂࿔᭿ࡡೳḾష⏕࡝࡜ᵕࠍ࡝⏍∸Ὡᛮࢅ♟ࡌࣚࢸ࢔ࢨࢤ࣭ࣜࡢࠉࡐࡡ࣏ࢠࣞࣚࢠࢹࣤᵋ㏸࠾ࡼ࣎ࣛࢢࢰ
࢕ࢺ࡛⩻࠻ࡼࡿࡾ࠿ࠉᮅࡓ⏍ྙᠺ㐿ఎᏄࡢྜྷᏽࡈࡿ࡙࠷࡝࠷ࠊࡐࡆ࡚ࠉࣚࢸ࢔ࢨࢤ࣭ࣜ⏍⏐Ⳟ࡚࠵ࡾ
Penicillium luteo-aurantium ࡻࡽ᩺ぞ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㸝PKS㸞㐿ఎᏄࡡࢠ࣭ࣞࢼࣤࢡࢅモࡲࡒࠊࡱࡍࠉ᪜
▩ࡡ⣊≟Ⳟ PKS 㐿ఎᏄࡡ⦨ྙ㓕⣪ KS ࢺ࣒࢕࡛ࣤ࢓ࢨࣜᇱ㌹⛛㓕⣪ AT ࢺ࣒࢕ࣤ㛣࡚ࡻࡂಕᏋࡈࡿ࡙࠷ࡾ࢓
࣐ࢿ㓗㒼าࢅࡵ࡛࡞ PCR ࣈࣚ࢕࣏࣭ࢅࢸࢧ࢕ࣤࡊࠉPKS ᩷∞ࡡቌᖕࢅ⾔ࡖࡒࠊT-࣊ࢠࢰ࣭࡞ࢦࣇࢠ࣭ࣞࢼࣤ
ࢡᚃࠉ㒼าỬᏽࡊࡒ࡛ࡆࢀࠉ㸨⛸ࡡ␏࡝ࡾ PKS ᩷∞࠿☔ヾࡈࡿࡒࠊࡐࡆ࡚ࠉ྘ PKS ᩷∞࠾ࡼࢣࢿ࣑࢚࣭ࢗ࢞
ࣤࢡࡊࠉධ㛏ሲᇱ㒼าࢅỬᏽࡊࡒࠊpla-1 ࡡࢤ࣭ࢺࡌࡾ PKS ࡢධ㛏 2332 aa ࡚ࠉࢢࢹ㑇ඔ㓕⣪ KR ࢺ࣒࢕ࣤࢅ
ࡵࡗࡆ࡛࠾ࡼࠉ㑇ඔᆵ PKS ࡛⩻࠻ࡼࡿࡒࠊpla-5 ࡢ㑇ඔᆵ PKS ࡡ C ᮆ࡞᭞࡞㟸ࣛ࣍ࢮ࣭࣑ᆵ࣋ࣈࢲࢺྙᠺ㓕⣪㸝NRPS㸞࠿⦽࠿ࡖࡒ PKS/NRPS ࣀ࢕ࣇࣛࢴࢺ㓕⣪㸝ධ㛏 5270 aa㸞ࢅࢤ࣭ࢺࡌࡾ㐿ఎᏄ࡛⩻࠻ࡼࡿࡒࠊࡱ
ࡒࠉpla-12 ࡢධ㛏 2049 aa ࡡⰶ㤮᪐ᆵ PKS ࢅࢤ࣭ࢺࡌࡾࡵࡡ࡛⩻࠻ࡼࡿࡒࠊpla-1 ࡞ࡗ࠷࡙ࠉࡐࡡᶭ⬗ࢅ☔ヾࡌࡾࡒࡴࠉධ㛏㐿ఎᏄࢅ Gateway Ἢࢅ⏕࠷࡙⣊≟Ⳟ⏕Ⓠ⌟࣊ࢠࢰ࣭pTAex3R ࡞ᑙථࡊࡒࠊᵋ⠇ࡊࡒⓆ⌟ࣈࣚࢪ࣐ࢺ pHS001 ࡚ Aspergillus oryzae ࢅᙟ㈻㌹ᥦࡊࠉㄇᑙᇰ㣬ᚃࠉ⏍⏐໩ྙ∸ࡡ HPLC ฦᯊࢅ⾔ࡖࡒࠊࡐࡡ⤎ᯕࠉᇰ㣬ᾦ୯࡞ࠉࢤࣤࢹ࣭ࣞࣜࡡᙟ㈻㌹ᥦమ࡞ࡢぜࡼࡿ࡝࠷≁␏Ⓩ࡝ࣅ࣭ࢠ࠿᳠ฝࡈࡿࠉ⌟ᅹࠉࡆࡡ໩ྙ∸ࡡ༟㞫᮪௲࡞ࡗ࠷࡙᳠ゞ୯࡚࠵ࡾࠊࡱࡒࠉ௙ࡡ PKS 㐿ఎᏄ࡞ࡗ࠷࡙ࡵࠉⓆ⌟ࣈࣚࢪ࣐ࢺࡡᵋ⠇ࢅ㐅ࡴ࡙࠷ࡾࠊ
Cloning and functional analysis of novel PKS genes from Penicillium luteo-aurantium
Hiroki Sano, Emi Ishii, Isao Fujii, Yutaka Ebizuka(Graduate School of Pharmaceutical Sciences, Univ. of Tokyo)

P-54⣊≟Ⳟⰶ㤮᪐࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ࡡࢠࣚ࢕ࢭࣤࢦ࢕ࢠ࣭ࣚࢭࢺ࣒࢕ࣤᐋᓧ ಆࠉ⸠஬ ໌ࠉᾇ⩹ሪ ㇇ 㸝᮶ኬ㝌࣬ⷾ㸞⣊≟Ⳟ Aspergillius fumigatus ࡡ࣒ࣚࢼࣤ⏍ྙᠺ࡞㛭ࢂࡾ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㸝PKS㸞࡚࠵ࡾ Alb1p ࡢࣉࣈࢰࢢࢰ࢕ࢺ࡚࠵ࡾࢻࣆࢹࣅࣞࣤ໩ྙ∸ YWA1 ࢅ⏍ᠺࡌࡾࠊྜྷࡋࡂ YWA1 ࡡྙᠺ㓕⣪࡚࠵ࡾ Aspergillus nidulans
ࡡ WA ࡞࠽࠷࡙ࠉࡐࡡ㸸ᮆ㡷ᇡ࠿ࢠࣚ࢕ࢭࣤᆵࡡཬᚺ࡞ࡻࡽ YWA1 ࡡ㸧⎌┘ࡡ㛚⎌ࢅり፳ࡌࡾࢺ࣒࢕࡚ࣤ࠵
ࡾࡆ࡛ࢅ඙࡞ሒ࿈ࡊࡒ࠿ࠉ௑ᅂࠉAlb1p ࡡ㓕ẍⓆ⌟⣌ࢅᵋ⠇ࡊࠉࡆࡿࢅ⏕࠷࡙ࢠࣚ࢕ࢭࣤࢦ࢕ࢠ࣭ࣚࢭ㸝CYC㸞
ࢺ࣒࢕ࣤࡡᶭ⬗よᯊࢅモࡲࡒࡡ࡚ሒ࿈ࡌࡾࠊࡱࡍࠉalb1 cDNA ࢅ㓕ẍⓆ⌟࣊ࢠࢰ࣭pYES-DEST52 ࡞ᑙථࡊࠉ
Ⓠ⌟ࣈࣚࢪ࣐ࢺ pYES-albY ࢅᵋ⠇ࡊࡒࠊࡆࡿࢅ࣌ࢪ࣌ࣂࣤࢷࢷ࢕ࣤ㌹⛛㓕⣪ NPGA ࡛࡛ࡵ࡞㓕ẍ࡚භⓆ⌟ࡈࡎࡾࡆ࡛࡞ࡻࡽࠉYWA1 ࠿⏍⏐ࡈࡿࡾࡆ࡛ࢅ☔ヾࡊࡒࠊୌ᪁ࠉAlb1p ࠾ࡼ CYC ࢺ࣒࢕ࣤࢅḖ኶ࡈࡎࡒን␏మ
Cd-3 ࡚ࡢࠉࣉࣈࢰࢢࢰ࢕ࢺ୯㛣మࡡ㸧⎌┘ࡡࢠࣚ࢕ࢭࣤ⎌໩ཬᚺ࠿㐅⾔ࡎࡍࠉ࢕ࢮࢠ࣏ࣛࣤమ DHCI ࠿⏍⏐ࡈࡿࡒࠊࡐࡆ࡚ࠉCYC ࢺ࣒࢕ࣤ㐿ఎᏄ࡛ Cd-3 㐿ఎᏄࢅࡐࡿࡑࡿ␏࡝ࡾ࣊ࢠࢰ࣭࡞ᑙථࡊࠉ஦⛸ࡡⓆ⌟ࣈࣚࢪ࣐ࢺࢅᵋ⠇ࡊࠉභ࡞ྜྷୌࡡ㓕ẍ࡚භⓆ⌟ࡈࡎࡒ࡛ࡆࢀࠉධ㛏 Alb1p ࡛ྜྷᵕ࡞ࠉYWA1 ࠿⏍ᠺࡌࡾ஥ࢅ☔ヾࡊࡒࠊࡆࡡ⤎ᯕࡢࠉCYC ࢺ࣒࢕ࣤ࠿≺❟ࡊࡒ㓕⣪࡛ࡊ࡙ Cd-3 ୕ࡡࣉࣈࢰࢢࢰ࢕ࢺ୯㛣మ࡞഼ࡀࠉࢠࣚ࢕ࢭ
ࣤ⎌໩ཬᚺࢅり፳ࡊࡒࡆ࡛ࢅ♟ࡊ࡙࠷ࡾࠊ⌟ᅹࠉ௙ࡡ⣊≟Ⳟⰶ㤮᪐ PKS ࡡ CYC ࢺ࣒࢕ࣤ࠿ྜྷᵕ࡞ᶭ⬗ࡌࡾ࠾࡜࠹࠾ࢅ☔ヾࡌࡾࡒࡴࠉPhoma BAUA2861 ᰬࡡࢿࢻࢢࢰ࢕ࢺ PKS ࡚࠵ࡾ PNK2ࠉཀྵࡦࠉColletotrichum
lagenarium ࡡ࣋ࣤࢰࢢࢰ࢕ࢺ PKS ࡚࠵ࡾ PKS1 ࡞ࡗ࠷࡙ࠉࡐࡿࡑࡿ CYC ࢺ࣒࢕ࣤࡡⓆ⌟ࣈࣚࢪ࣐ࢺࢅᵋ⠇ࡊࠉ
Cd-3 ࡛භⓆ⌟ࡈࡎࡾ஥࡞ࡻࡽࠉࣉࣈࢰࢢࢰ࢕ࢺ୯㛣మࡡࢠࣚ࢕ࢭࣤ⎌໩㓕⣪࡛ࡊ࡙ᶭ⬗ࡊ࡙ YWA1 ࠿⏍⏐ࡈࡿࡾ࠾࡜࠹࠾ࢅ᳠ゞࡊ࡙࠷ࡾࠊࡱࡒࠉAlb1p ࡡ CYC ࢺ࣒࢕ࣤࡡኬ⭘Ⳟ࡚ࡡⓆ⌟ࠉ⢥⿿࡞ࡗ࠷࡙ࡵ᳠ゞ୯࡚࠵ࡾࠊStudies on the Claisen cyclase domains of fungal aromatic polyketide synthases
Shun Miyazaki, Isao Fujii㸡Yutaka Ebizuka(Graduate School of Pharmaceutical Sciences, Univ. of Tokyo)

77

P-55
ⰶ㤮᪐⧖㏁ࡊᆵࢰ࢕ࣈϨ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪ࡡⓆ⌟࡛ᶭ⬗よᯊ᲻ཾᬓ⨶㸡 ⸠஬ ໌㸡 ᾇ⩹ሪ㇇ 㸝᮶ኬ㝌ⷾ㸞⣊≟Ⳟࡡ࣎ࣛࢢࢰ࢕ࢺྙᠺ㓕⣪㸝PKS㸞ࡢࠉࡐࡡ࡮࡛ࢆ࡜࠿ࢰ࢕ࣈϨ࡞ฦ㢦ࡈࡿࡾ࠿ࠉࣁࢠࢷࣛ࢓࡞ぜࡼࡿࡾ࣓ࢩ࣭ࣖࣚᆵ࡚ࡢ࡝ࡂࠉ྘Ὡᛮ୯ᚨ࠿⧖㏁ࡊ࡙ཬᚺ࡞㛭୙ࡌࡾ iterative ᆵ࡚࠵ࡾࠊ࡝࠾࡚ࡵ 6-࣒ࢲࣜࢦࣛࢲࣜ㓗㸝6MSA㸞ྙᠺ㓕⣪㸝MSAS㸞ࡢࠉ⣊≟Ⳟࡡࢰ࢕ࣈϨPKS ࡛ࡊ࡙᭩ᑚ࡚࠵ࡾࡒࡴࠉࡐࡡᵋ㏸࡛ཬᚺᶭ
ᵋࢅよ᪺ࡌࡾࡒࡴࡡࡻ࠷ࢰ࣭ࢣࢴࢹ࡚࠵ࡾ࡛⩻࠻ࡼࡿࡾࠊ
Aspergillus terreus ࡻࡽࢠ࣭ࣞࢼࣤࢡࡊࡒ MSAS 㐿ఎᏄ atX (Mol. Gen. Genet. 253 1-10(1996))ࢅ Bacillus
subtilis ࡡ࣌ࢪ࣌ࣂࣤࢷࢷ࢕ࣤ㌹⛛㓕⣪ Sfp ࡛࡛ࡵ࡞㓕ẍࢅᐙ୹࡛ࡊ࡙Ⓠ⌟ࡈࡎࠉ6MSA ࡡ⏍⏐࡞ࡻࡽ ATX ࠿㓕ẍ࡞࠽࠷࡙ᶭ⬗ࡌࡾࡆ࡛ࢅ☔ヾࡊࡒࠊୌ᪁ࠉධ㛏 ATX ࡡ N ᮆഁ⦨ྙ㓕⣪ࢺ࣒࢕ࣤࡡୌ㒂ࡱ࡚ࠉࡱࡒ C ᮆ
ഁࡡ 9 ࢓࣐ࢿ㓗ࢅࡐࡿࡑࡿெⅥⓏ࡞Ḗ኶ࡈࡎࡒ ATX ን␏మࡢᏰධ࡞ᶭ⬗ࢅ኶࠷ࠉ6MSA ࡡ⏍⏐ࡢࡲࡼࡿ࡝࠾
ࡖࡒࠊ࡛ࡆࢀ࠿ࠉ༟≺࡚ࡢ୘Ὡᛮ࡝ N ᮆḖ኶మ࡛ C ᮆḖ኶మࢅ㓕ẍ࡞࠽࠷࡙භⓆ⌟ࡈࡎࡒ࡛ࡆࢀࠉᙟ㈻㌹ᥦ
మࡢ 6MSA ࢅ⏍⏐ࡊࠉࡐࡡ 6MSA ⏍⏐ᛮࡢධ㛏ᙟ㈻㌹ᥦమࡡ⣑ 1/20 ࡚࠵ࡖࡒࠊࡆࡡ⤎ᯕࡢࠉN ᮆࡡḖ኶࡞ࡻࡽ୘Ὡᛮ࡛࡝ࡖࡒ⦨ྙ㓕⣪ࢺ࣒࢕ࣤࢅࡵࡗࢦࣇࣗࢼࢴࢹࠉ࠽ࡻࡦࠉC ᮆ❻ࡡḖ኶࡞ࡻࡽ୘Ὡᛮ࡛࡝ࡖࡒ࢓ࢨ
࣭ࣜ࢞ࣔࣛࣕࣈࣞࢷ࢕ࣤࢅࡵࡗࢦࣇࣗࢼࢴࢹ࠿┞பష⏕ࡌࡾࡆ࡛࡞ࡻࡽࠉATX ࡡ 6MSA ྙᠺり፳ᶭ⬗࠿්ᵋᠺࡈࡿࡒࡵࡡ࡛⩻࠻ࡼࡿࠉATX ࡡὩᛮ୯ᚨ࠿ᑛ࡝ࡂ࡛ࡵ㸧ࡗࡡࢦࣇࣗࢼࢴࢹࡡᶭ⬗ࢺ࣒࢕ࣤ࡞ࡻࡽᵋᠺࡈࡿ
࡙࠷ࡾࡆ࡛ࢅ♟ࡊ࡙࠷ࡾࠊ⌟ᅹࠉࡻࡽレ⣵࡝ࢦࣇࣗࢼࢴࢹ-ࢺ࣒࢕ࣤ㛣ࡡ┞பష⏕࡞ࡗ࠷࡙᳠ゞ୯࡚࠵ࡾࠊ
Expression and Functional Analysis of Aromatic Iterative Type I Polyketide Synthases
Tomomi Moriguchi, Isao Fujii, Yutaka Ebizuka(Graduate School of Pharmaceutical Sciences, Univ. of Tokyo)

P-56⣊≟Ⳟ A. nidulans ࢓࣐࣭ࣚࢭ㐿ఎᏄ⩄ࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣛࣤࢡ
୯ᮟ㈏㸡∶⏛ᬓ⿩㸡ຊ⸠㞖ኃ㸡ᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞
Aspergillus ᒌ࡞࠽ࡄࡾ࢓࣐࣭ࣚࢭ㐿ఎᏄ⩄ࡡⓆ⌟ࡢࠉᇱ㈻࡚࠵ࡾࢸࣤࣈࣤࡷࡐࡡฦよ⏐∸࣏ࣜࢹ࣭ࢪ࡝࡜
࡞ࡻࡽㄇᑙࡈࡿࡾࠊࡆࡿࡱ࡚࡞ࠉA. nidulans ࡞࠽࠷࡙ agdA ࠉagdB ࡷ amyB ࡝࡜ࡡ࢓࣐࣭ࣚࢭ㐿ఎᏄ⩄ࡡ
⏍⌦ᏕⓏㄇᑙ∸㈻ࡢࠉ࢕ࢮ࣏ࣜࢹ࣭ࢪ࡚࠵ࡽࠉࡆࡡㄇᑙࢨࢡࢻࣜ࠿㌹෕Ὡᛮ໩ᅄᏄ AmyR ࡞ఎ㐡ࡈࡿࠉ㌹෕࠿Ὡᛮ໩ࡈࡿࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖ࡙࠷ࡾࠊࡊ࠾ࡊࠉࡌ࡬࡙ࡡ࢓࣐࣭ࣚࢭ㐿ఎᏄ࠿ྜྷᵕࡡᶭᵋ࡚㌹෕Ὡᛮ໩ࡈࡿࡾ࠾࡞ࡗ࠷࡙ࡢࠉᮅࡓ᪺ࡼ࠾࡞࡝ࡖ࡙࠷࡝࠷ࠊᮇ◂✪࡚ࡢࠉA. nidulans ࢣࢿ࣑୕࡞Ꮛᅹࡌࡾࡌ࡬࡙ࡡ࢓
࣐࣭ࣚࢭ㐿ఎᏄࢅྜྷᏽࡊࠉࡐࡡⓆ⌟ࣈࣞࣆ࢒࢕ࣜࢅ⥑⨮Ⓩ࡞よᯊࡊࡒࠊA. nidulans genome database ࠾ࡼࠉfamily13ࠉ15ࠉ31 ࡞ᒌࡌࡾ glycosyl hydrolase ࢅ᢫ฝࡊࠉࡈࡼ࡞ manual
annotation ࡞ࡻࡽ࢓࣐࣭ࣚࢭ㐿ఎᏄࢅྜྷᏽࡊࡒࠊࡐࡡ⤎ᯕࠉȘ-࢓࣐࣭ࣚࢭ 7 ⛸㢦ࠉȘ-ࢡࣜࢤࢨࢱ࣭ࢭ 7 ⛸㢦ࠉࢡࣜࢤ࢓࣐࣭ࣚࢭ 2 ⛸㢦ࡡᏋᅹ࠿᪺ࡼ࠾࡛࡝ࡖࡒࠊ༖ᏽ㔖Ⓩ RT-PCR ࢅ⾔࠷ࠉࡆࡿࡼ࢓࣐࣭ࣚࢭ㐿ఎᏄ⩄ࡡ
Ⓠ⌟ࣂࢰ࣭ࣤࢅ⥑⨮Ⓩ࡞よᯊࡊࡒ⤎ᯕࠉ࢕ࢮ࣏ࣜࢹ࣭ࢪ࡞ᚺ➽ࡊ࡙Ⓠ⌟࠿ㄇᑙࡈࡿࡾ㐿ఎᏄ⩄࠿Ꮛᅹࡌࡾ࠿ࠉ
࣏ࣜࢹ࣭ࢪ࡞ࡻࡽㄇᑙࡈࡿࡾ㐿ఎᏄࡢᏋᅹࡊ࡝࠾ࡖࡒࠊࡱࡒࠉ⅛⣪″㣒㣱᮪௲ୖ࡞࠽࠷࡙ࠉ࢓࣐࣭ࣚࢭ㐿ఎᏄ⩄ࡢ AmyR 㟸౪ᏋⓏ࡝Ⓠ⌟ㄇᑙࢅ♟ࡊࡒࠊ⅛⣪″㣒㣱᮪௲ୖ࡞࠽ࡄࡾⓆ⌟ㄇᑙࡢࠉ࣭࢜࣍ࣤ࢜ࢰ࣍ࣚ࢕ࢹ
ࣛࣈࣝࢴࢨࣘࣤ࠾ࡼࡡ⬲ᢒโ࡚࠵ࡾྊ⬗ᛮ࠿࠵ࡾࡒࡴࠉ⌟ᅹࠉ㌹෕ᢒโᅄᏄ CreA ࡡን␏ᰬࢅ⏕࠷ࡒよᯊࢅ⾔ࡖ࡙࠷ࡾࠊExpression profiling of amylase genes in Aspergillus nidulans
Takashi Nakamura, Tomohiro Makita, Masashi Kato, Tetsuo Kobayashi(Dept. of Biological Mechanisms and Functions, Nagoya Univ.)

78

P-57
㯔Ⳟ࢞ࢨࣚࢻ࣭ࢭ࣬ࢬ࣭ࣜࣚࢭㄇᑙᅄᏄ AoXlnR ࡡ㌹෕โᚒᶭᵋ
㔕ཾ♰ྒྷ㸡⏛୯ᑋᇱ㸡ຊ⸠㞖ኃ㸡ᑚᯐူኰ 㸝ྞኬ㝌⏍࿤㎨࣬⏍∸ᶭᵋ㸞
ࠜ┘Ⓩࠝ AoXlnR ࡢ Aspergillus oryzae ࡡ࢞ࢨࣚࢻ࣭ࢭ࣬ࢬ࣭ࣜࣚࢭ㐿ఎᏄ⩄ࡡⓆ⌟ࢅโᚒࡌࡾ㌹෕ᅄᏄ࡚࠵ࡾ㸣ᮇᅄᏄࡢ࢞ࢨࣚࣤࡷ࢞ࢨ࣭ࣞࢪ࡞ᚺ➽ࡊ࡙㸡ᨥ㒼ୖ㐿ఎᏄ⩄ࡡ㌹෕ࢅὩᛮ໩ࡌࡾ㸣ࡊ࠾ࡊ㸡 AoXlnR ࡡㄇᑙࢨࢡࢻࣜヾㆉᶭᵋ㸡㌹෕โᚒᶭᵋࡢ᪺ࡼ࠾࡛࡝ࡖ࡙࠷࡝࠷㸣௑ᅂ㸡 AoXlnR ࡞ࡻࡾ㌹෕โᚒᶭᵋࡡよ᪺ࢅ┘Ⓩ࡛ࡊ㸡㟸ㄇᑙ᮪௲㸡ㄇᑙ᮪௲ୖ࡚ AoXlnR ࡡよᯊࢅ⾔ࡖࡒࡡ࡚ሒ࿈ࡌࡾ㸣ࠜ᪁Ἢ࣬⤎ᯕࠝ GFP-AoXlnR ⼝ྙ㐿ఎᏄࢅ A. oryzae AoXlnR ◒ቪᰬ࡞ᑙථࡊࡒ㸣◒ቪᰬ࡞࠽ࡄࡾ࢞ࢨࣚࢻ
࣭ࢭㄇᑙ⬗ࡡḖ኶ࡢ GFP-AoXlnR 㐿ఎᏄࡡᑙථ࡞ࡻࡽᅂᚗࡊࡒࡒࡴ㸡 GFP-AoXlnR ࡛㔕⏍ᆵ AoXlnR ࡢྜྷ➴ࡡᶭ⬗ࢅᣚࡗ࡛⩻࠻ࡼࡿࡾ㸣 GFP-AoXlnR ࡡ⣵⬂හᑻᅹᛮࢅ࢞ࢨ࣭ࣞࢪ῟ຊ㸡㟸῟ຊ᮪௲࡚ふᐳࡊࡒ⤎ᯕ㸡
GFP-AoXlnR ࡢᖏ࡞ᰶ࡞ᑻᅹࡊ࡙࠷ࡒ㸣ᚉࡖ࡙㸡 AoXlnR ࡢᰶහ࡚ㄇᑙࢨࢡࢻࣜࢅུᐖࡊ࡙Ὡᛮ໩ࡌࡾ࡛⩻࠻ࡼࡿࡾ㸣ࡱࡒ㸡 myc-AoXlnR ⼝ྙ㐿ఎᏄࢅ A. oryzae ࡞ᑙථ㸡࢞ࢨ࣭ࣞࢪ῟ຊ㸡㟸῟ຊ᮪௲ࡡⳞమࡻࡽ⣵⬂᢫ฝᾦࢅㄢ⿿ࡊ㸡ᢘ myc ᢘమࢅ⏕࠷࡙࢙ࢗࢪࢰࣤࣇࣞࢴࢷ࢔ࣤࢡよᯊࢅ⾔ࡖࡒ㸣ࡐࡡ⤎ᯕ㸡 myc-AoXlnR ࡡ
SDS-PAGE ୕ࡡ⛛ິᗐ࠿࢞ࢨ࣭ࣞࢪ῟ຊ᮪௲ୖ࡚఩ୖࡌࡾࡆ࡛࠿᪺ࡼ࠾࡛࡝ࡖࡒ㸣ࡆࡡ⛛ິᗐࡡ఩ୖࡢ࢞ࢨ
࣭ࣞࢪ≁␏Ⓩ࡞ᘤࡀ㉫ࡆࡈࡿ㸡ࡱࡒ㸡࢞ࢨ࣭ࣞࢪࡡ㝎ཡ࡞ࡻࡽᅂᚗࡊࡒ㸣௧୕ࡡ⤎ᯕࡢ㸡࢞ࢨ࣭ࣞࢪ࡞ᚺ➽ࡊ࡙㸡 AoXlnR ࡞రࡼ࠾ࡡྊ㏣Ⓩಞ㣥ࡱࡒࡢᵋ㏸ን໩࠿㉫ࡆࡾࡆ࡛ࢅណ࿝ࡊ࡙࠷ࡾ㸣Regulatory mechanism of AoXlnR, a transcriptional activator of xylanase and cellulose genes in Aspergillus
oryzae.
Yuji Noguchi, Hisaki Tanaka, Masashi Kato, Tetsuo Kobayashi(Dept. of Biological Mechanisms and Functions, Graduate School of Bioagricultural Sciences, Nagoya University)

P-58
㯔Ⳟ CCAAT ⤎ྙᅄᏄ ඁເ㜴ᐐᆵን␏ࢦࣇࣗࢼࢴࢹࡡよᯊ㸯โᚒࡈࡿࡾ㐿ఎᏄ ⩄ࡡ⥑⨮Ⓩよᯊ࡫ࡡฺ⏕ᮙᒜ⣟ஒࠉ㧏ᶣ᪺⌌ࠉྙ⏛⚵▦ࠉᑚᯐူኰࠉຊ⸠㞖ኃ ྞኬ㝌࣬⏍࿤㎨࣬⏍∸ᶭᵋ 
糸状菌 CCAAT 結合因子は Hap 複合体とも呼ばれ、 HapB/C/E の 3 つのサブユニットからなり、多くの産業上重要な酵素遺伝子の転写を促進している。これまでに Hap 複合体の性質は詳細に解析されてきているが、どのような遺伝子群が制御されているのかという全体像は依然として明らかではない。モデル糸状菌
Aspergillus nidulans の解析から Hap サブユニットの遺伝子欠失株は生育が遅く胞子の着生が悪くなることが明らかになっており、麹菌においての欠失株の取得も容易ではないと考えられる。多核である麹菌では欠失株の分離はさらに難しく、現在までのところ成功していない。今回、マイクロアレイによる網羅的解析に必要なサブユニット遺伝子欠失株に替わる優勢阻害型（ドミナントネガティブ）変異株の取得を試みた。
Hap サブユニットは種間で高度に配列が保存された領域を有している。保存領域内の推定 DNA 結合ドメインを変異させた HapB ( HapB-1M ) 遺伝子を野性型 A. nidulans に導入したところ、 Hap 複合体依存であることが既に明らかになっているアミラーゼやエンド-β-グルカナーゼ A 活性の低下が明らかとなった。これらの現象は、変異 HapB が正常な HapB から他のサブユニット HapC/E を奪うことにより起こる優勢阻害効
果に起因していることが示唆された。麹菌に HapB-1M 遺伝子を導入した株でも同様の優性阻害効果が認められた。現在、この株を用いてマイクロアレイ解析に向けた条件検討を進めている。
Analysis on a dominant-negative mutant subunit of the Aspergillus oryzae CCAAT-box binding factor:
applications for the comprehensive analysis of genes controlled by the factor.
Junya Sugiyama, Akemi Takahashi, Hideya Goda, Tetsuya Kobayashi, Masashi Kato
(Dept. of biological mechanisms and functions, Grad. Sch. of Nagoya Univ.)

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㯜㯔Ⳟ Aspergillus oryzae ࡡ HSP30 㐿ఎᏄࣈ࣓࣭ࣞࢰ࣭ࡡ Deletion よᯊᮿୖ┷⏜⨶ࠉ㕝ᮄ ⪵ࠉᴃᮇ៿ୌࠉ᯵ᮄ ㇇ 㸝㸝≺㸞㣏⥪◂㸞[┘ Ⓩ ] 㯔 Ⳟ ࡞ ࠽ ࡄ ࡾ ∸ ㈻ ⏍ ⏐ ࡞ ࠽ ࠷ ࡙ ᙁ ງ ࠾ ࡗ ᭯ ⏕ ࡝ 㐿 ఎ Ꮔ Ⓠ ⌟ ࣈ ࣞ ࣓ ࣭ ࢰ ࣭ ࡡ 㛜 Ⓠ ࠿ ᭻ ࡱࡿ ࡙ ࠷ ࡾ ࠊ ᮇ ◂ ✪ ࡚ ࡢ ࠉ ࡐ ࡡ ୌ ⎌ ࡛ ࡊ ࡙ ࠉ 㯜 㯔 Ⳟ Aspergillus oryzae ࡡ 㧏 Ὼ ㄇ ᑙ ᛮ 㐿 ఎ Ꮔ ࡡ ࡥ ࡛ࡗ ࡚ ࠵ ࡾ HSP30 㐿 ఎ Ꮔ 㸝 1 㸞 ࡞ ╌ ┘ ࡊ ࠉ ࡐ ࡡ ࣈ ࣞ ࣓ ࣭ ࢰ ࣭ よ ᯊ ࢅ ⾔ ࡖ ࡒ ࠊ[᪁ Ἢ ࡛ ⤎ ᯕ ] HSP30 㐿 ఎ Ꮔ ࡡ Ᏸ ධ 㛏 cDNA ཀྵ ࡦ ࠉ ᮇ 㐿 ఎ Ꮔ ࡡ 5v୕ Ὦ ᇡ ⣑ 4kb ࢅ ཱི ᚋ ࡊ ࠉ ࡆࡡ 㡷 ᇡ ࡡ 㒼 า ࢅ よ ᯊ ࡊ ࡒ ࡛ ࡆ ࢀ ࠉ5v୕ Ὦ ᇡ ࡞ ࡢ ࢪ ࢹ ࣝ ࢪ ᚺ ➽ 㒼 า ࡞ 㢦 జ ࡡ ሲ ᇱ 㒼 า ࠿ 々 ᩐ Ꮛ ᅹࡌ ࡾ ࡆ ࡛ ࠿ よ ࡖ ࡒ ࠊ ḗ ࡞ ࠉ ஢ ῼ ⩳ ゼ 㛜 ጙ Ⅴ ࡻ ࡽ ୕ Ὦ ⣑ 2㹠 b ࡡ ࣈ ࣞ ࣓ ࣭ ࢰ ࣭ 㡷 ᇡ ࢅ ⣑ 500 ཧ ࡢ
100bp ࡡ 㛣 㝰 ࡚ ㏪ ḗ D eletion ࡊ ࠉ ࣈ ࣞ ࣓ ࣭ ࢰ ࣭ ࢓ ࢴ ࢬ ࢕ ⏕ ࣊ ࢠ ࢰ ࣭ pNAG M(pNAGT4㸝 ኬ 㛭 ( ᰬ )ࡻ ࡽ ᜠ ୙ 㸞 ࡻ ࡽ ᨭ ን )࡞ ⤄ ࡲ ㎲ ࡲ 13 ⛸ 㢦 ࡡ ᙟ ㈻ ㌹ ᥦ ᰬ ࢅ ష ᠺ ࡊ ࡒ ࠊ ࡐ ࡊ ࡙ GUS Ὡ ᛮ ࡞ ᑊ ࡌ ࡾ ᫤㛣 ࡛ Ὼ ᗐ ን ໩ ࡡ ᙫ 㡢 ࢅ ᳠ ゞ ࡊ ࡒ ࠊ ྘ Deletion ᰬ ࢅ YPD ᾦ మ ᇰ ᆀ ࡚ 30Υ ୌ ᬄ ᇰ 㣬 ᚃ ࠉ GUS Ὡ ᛮࢅ ῼ ᏽ ࡊ ࡒ ࡛ ࡆ ࢀ ࠉ -2045㹳 -476 㡷 ᇡ ࢅ ๎ 㝎 ࡊ ࡒ ᫤ ࡛ Ẓ ࡬ ࡙ ࠉ -388 ࡱ ࡚ ๎ ࡾ ࡛ ኬ ࡀ ࡂ ఩ ୖ ࡊ ࡒ ࠊࢿ ࣭ ࢧ ࣤ よ ᯊ ࡡ ⤎ ᯕ ࡚ ࡵ ࠉ30Υ ᇰ 㣬 ᫤ ࡡ ㌹ ෕ Ὡ ᛮ ࠿ ୖ ࠿ ࡖ ࡙ ࠷ ࡒ ࠊࡆ ࡡ -476㹳 -388 ࡞ ࡢ ࠉCCA AT㒼 า ࠿ Ꮛ ᅹ ࡊ ࡙ ࠷ ࡒ ࠊ-2045㹳 -388 ࡱ ࡚ ๎ 㝎 ࡊ ࡒ ྘ ᰬ ࡢ ࠉ40Υ 2 ᫤ 㛣 ࡡ 㧏 Ὼ ㄇ ᑙ ฌ ⌦ ࡞ ࡻ ࡾ GU SὩ ᛮ ࡡ ቌ ຊ ࠿ ぜ ࡼ ࡿ ࡒ ࠿ ࠉ-285 ࡱ ࡚ ๎ 㝎 ࡌ ࡾ ࡛ ࠉ30Υ ᇰ 㣬 ᫤ ࡓ ࡄ ࡚ ࡝ ࡂ ࠉ㧏 Ὼ ㄇ ᑙ ࡚ ࡵ GUS Ὡᛮ ࠿ ⴥ ࡊ ࡂ ఩ ୖ ࡊ ࡒ ࠊ-388㹳 -285 㡷 ᇡ ࡞ ࡢ ࠉ㸽 㹈 㸺 ࡛ ࢪ ࢹ ࣝ ࢪ ᚺ ➽ 㒼 า ࡞ 㢦 జ ࡡ 㒼 า ࠿ Ꮛ ᅹ ࡊ ࠉࡆࡿ࠿㧏Ὼㄇᑙ࡞㛭ࢂࡖ࡙࠷ࡾ࡛⩻࠻ࡼࡿࡾࠊ
(1) ᮿ ୖ ࡼ ࠉ ᪝ ᮇ ㎨ ⰹ ໩ Ꮥ ఌ  2005 ᖳ ᗐ ኬ ఌ ㅦ ⁿ さ ᪠ 㞗 ࠉ 㹥 232
Analysis of a promoter region of HSP30 gene from Aspergillus oryzae and expression of GUS gene
Mayumi Matsushita, Satoshi Suzuki, Ken-Ichi Kusumoto, Yutaka Kashiwagi
㸝Natl. Food Res. Inst.)

P-60
㯔Ⳟ Aspergillus oryzae ࡡ఩Ὼㄇᑙࣈ࣓࣭ࣞࢰ࣭ࡡ༟㞫஁⏛༡ඔ㸡ላᾀᏄ㸡⛑Ὂ஦㸡Ꮽ㒂ᗛ஁㸝᭮᰿෗⥪◂㸞㯔Ⳟ Aspergillus oryzae ࡢఎ⤣Ⓩ࡝㔂㏸⏐ᴏ࡚࠵ࡾΰ㒿ࠉ㓲ἔࠉ࿝ძ࡞౐⏕ࡈࡿ࡙࠷ࡾ㔔さ࡝᚜⏍∸࡚࠵ࡾࠊࡱࡒ㯔Ⳟࡢᵕࠍ࡝㓕⣪ࢅኣ㔖࡞ฦἢࡌࡾࡆ࡛࠿▩ࡼࡿࠉ㐿ఎᏄᕝᏕᡥἪࢅ⏕࠷࡙᭯⏕ࢰࣤࣂࢠ㈻⏍⏐࠿⾔ࢂࡿ࡙࠷ࡾࠊࢰࣤࣂࢠ㈻ࡡ⏍⏐࡞࠽࠷࡙㔔さ࡛࡝ࡾࡡࡢࠉ1.ࣈ࣓࣭ࣞࢰ࣭㐽ᢝࠉ2.ᇰ㣬᮪௲ࡡࢤࣤࢹ࣭࡚ࣞࣜ࠵
ࡾࠊᠻࠍࡢࡆࡿࡱ࡚࡞ࠉᾦమᇰ㣬࡚㧏Ⓠ⌟ࡌࡾ sodMࠉmelO ࡛ catB ࣈ࣓࣭ࣞࢰ࣭ࠉᅖమᇰ㣬࡚㧏Ⓠ⌟ࡌࡾ glaBࣈ࣓࣭ࣞࢰ࣭ࢅཱིᚋࡊࡒࠊࡊ࠾ࡊࡆࡿࡼࡡ㧏Ⓠ⌟ࣈ࣓࣭ࣞࢰ࣭ࢅฺ⏕ࡊ࡙ࡵࠉධ࡙ࡡ┘Ⓩࢰࣤࣂࢠ㈻࠿ኬ㔖⏍⏐࡚ࡀࡾࢂࡄ࡚ࡢ࡝࠷ࠊ⩳ゼᚃࡡࢰࣤࣂࢠ㈻ࡡࣆ࢚࣭ࣜࢸ࢔ࣤࢡࡷࣈࣞࢷ࢓࣭ࢭฦよ࡝࡜࠿ࠉࡐࡡཋᅄ࡛
ࡊ࡙ᣪࡅࡼࡿ࡙࠷ࡾࠊୌ᪁ኬ⭘Ⳟ࡞ࡻࡾࢰࣤࣂࢠ㈻⏍⏐࡞࠽࠷࡙ࡢࠉ఩Ὼ࡚ᇰ㣬ࡌࡾࡆ࡛࡞ࡻࡽᠺ⇅ࢰࣤࣂࢠࡡ཭㔖࠿ྡྷ୕ࡌࡾࡆ࡛࠿ሒ࿈ࡈࡿ࡙࠷ࡾࠊࡐࡆ࡚ᮇ◂✪࡚ࡢࠉ㯔Ⳟࡡ఩Ὼ࡚ࡡࢰࣤࣂࢠ㈻⏍⏐ࢨࢪࢷ࣑ࢅ
᳠ゞࡌࡾࡒࡴࠉ఩Ὼ࡚ᙁງ࡞Ⓠ⌟ㄇᑙࡌࡾࣈ࣓࣭ࣞࢰ࣭ࡡཱིᚋࢅモࡲࡒࠊࡱࡍࠉ㯔Ⳟ OSI1013 ᰬࢅ఩Ὼ࡛㏳ᖏࡡῺᗐ࡚ࣆࢪ࣏ᇰ㣬ࢅ⾔࠷ࠉ2 ᪝㛣ᇰ㣬ᚃ࡞྘Ⳟమ࠾ࡼ mRNA ࢅ⢥⿿ࡊࠉᏽἪ࡞ᚉ࠷ DNA ࣏࢕ࢠࣞ࢓ࣝ࢕よᯊࢅ⾔ࡖࡒࠊDNA ࣏࢕ࢠࣞ࢓ࣝ࢕よᯊࡡ⤎ᯕ࠾ࡼࠉ఩Ὼ࡚Ⓠ⌟㔖࠿ኣ࠷㐿ఎᏄ⩄ࢅ᢫ฝࡊࡒࠊ᢫ฝࡊࡒ㐿ఎᏄ⩄ࡡ୯࠾ࡼࠉ఩Ὼ࡚≁␏Ⓩ࡞Ⓠ⌟ㄇᑙࡈࡿࡾ 3 ⛸㢦ࡡ㐿ఎᏄࢅ㐽ᢝࡊࡒࠊ྘㐿ఎᏄࡡࣈ࣓࣭ࣞࢰ࣭㡷ᇡࢅࢦࣇࢠ࣭ࣞࢼࣤࢡࡊࠉGUS ࣭ࣝ࣎ࢰ࣭よᯊࢅ⾔ࡖࡒࠊⓆ⌟ࣈࣞࣆ࢒࢕ࣜࢅ᳠ゞࡊࡒ⤎ᯕࠉࡐࡡ࠹ࡔࡡ 2 ⛸㢦ࡡࣈ࣓࣭ࣞࢰ࣭ࡢ఩Ὼࣆࢪ࣏ᇰ㣬࡚≁␏Ⓩ࡞Ⓠ⌟ࡌࡾࡆ࡛࠿ึ᪺ࡊ
ࡒࠊ
Isolation and analysis of promoter regions required for low temperature induction in
Aspergillus oryzae
Hiromoto Hisada, Hiroko Tsutsumi, Yoji Hata, Yasuhisa Abe (Gekkeikan Sake Co. Ltd.)

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㯔Ⳟ࡛㓕ẍ࡞࠽ࡄࡾ㐿ఎᏄⓆ⌟ࡡẒ㍉よᯊᒜ᰷೒Ꮔ㸡ᑈᯐ㟱ᐃ㸡ఫ㔕ඔ᫓*㸡㧏℡஁⨶Ꮔ㸡ኬ⟺ಘୌ*ࠉ⏣⏛㞖஄ 㸝⏐⥪◂࣬*㔘Ἁᕝኬ㸞
㸱┘Ⓩ㸳㯔Ⳟ Aspergillus oryzae ࡢࠉ㒿࣬࿝ძ࣬㓲ἔ࡝࡜ࡡఎ⤣Ⓩ⏐ᴏࢅิࡴ࡛ࡊ࡙ࠉ㎾ᖳ࡚ࡡ㐿ఎᏄᕝᏕᢇ
⾙ࢅ⏕࠷ࡾ㓕⣪⏍⏐࡝࡜ࠉ᪝ᮇࡡࣁ࢕࢛ࢷࢠࢿࣞࢩ࣭⏐ᴏ࡞ᗀࡂ⏕࠷ࡼࡿ࡙࠷ࡾࠊ㎾ᖳࠉ㯔Ⳟࡡࢣࢿ࣑よᯊ࠿࡮࡯Ᏸ஡ࡊࠉ㯔Ⳟࡡᣚࡗ㐿ఎᏄࡡධㇲ࠿᪺ࡼ࠾࡞࡝ࡖࡒࠊࡐࡆ࡚ᠻࠍࡢࠉ㯔Ⳟࡡࢣࢿ࣑ሲᇱ㒼า࠾ࡼ஢ῼ
ࡈࡿࡒ㐿ఎᏄࡡ࡮࡯ධ࡙ࢅᦒ㍍ࡌࡾ࢛ࣛࢥ DNA ࣏࢕ࢠࣞ࢓ࣝ࢕ࢅషᠺࡊࡒࠊ᪜࡞ࠉࡆࡡ࣏࢕ࢠࣞ࢓ࣝ࢕ࢅ⏕࠷࡙ࠉᵕࠍ࡝ᇰ㣬᮪௲ࡷᙟ㈻㌹ᥦమࢅ⏕࠷ࡒよᯊࢅ㛜ጙࡊ࡙࠽ࡽࠉ㯔Ⳟࡡධ㐿ఎᏄࡡⓆ⌟᝗ሒ࡞㛭ࡌࡾኬぞᶅ࡝ࢸ࣭ࢰ࣭࣊ࢪࢅᵋ⠇ࡊࡗࡗ࠵ࡾࠊࡆࡡ᝗ሒᇱ┑ࡢࠉ㯔Ⳟࡡ㌹෕โᚒࢾࢴࢹ࣭࣠ࢠࢅよࡂࡒࡴ࡞㔔さ࡝㘵࡞࡝ࡾ࡛⩻࠻ࡼࡿࡾ 1)ࠊ௑ᅂࠉᠻࠍࡢࠉ୕オࡡ㯔Ⳟ DNA ࣏࢕ࢠࣞ࢓ࣝ࢕ࢅ⏕࠷࡙ࠉ࣓ࢸࣜ⏍∸࡚࠵ࡾฝⰾ㓕ẍ Saccharomyces cerevisiae ࡛ࡡ㐿ఎᏄⓆ⌟ࣈࣞࣆ࢒࢕ࣜࡡẒ㍉ࢅモࡲࡒࠊ㸱᪁Ἢ࣬⤎ᯕ㸳࠵ࡼ࠾ࡋࡴᇰ㣬ࡊࡒ㯔ⳞⳞమ࡞ Heat shock ฌ⌦ࢅ⾔࠷ࠉRNA ࢅ᢫ฝࡊ࡙࣏࢕ࢠࣞ࢓ࣝ࢕⏕⺧කᵾㆉ mRNA ࢅష⿿ࡊࡒࠊࡆࡡモᩩࢅ㯔Ⳟࡡ 11,000 㐿ఎᏄࡡࣈ࣭ࣞࣇࢅࢪ࣎ࢴࢹࡊࡒ࢛ࣛࢥ࢓ࣝ࢕ࢅ⏕࠷࡙
㐿ఎᏄࡡⓆ⌟よᯊࢅ⾔ࡖࡒࠊࡱࡒࠉ㐿ఎᏄⓆ⌟ࢸ࣭ࢰ࣭࣊ࢪ୕࡞࠵ࡾࠉ㓕ẍࡡྜྷᵕࡡᇰ㣬᮪௲ୖ࡚ࡡよᯊ⤎ᯕ࡛ࡡẒ㍉ࢅ⾔ࡖࡒࠊ㯔Ⳟ࠿᭯ࡌࡾ௥ㅨࣂࢪࢗ࢘࢕ࢅྱࡴ࡙ࠉHeat shock ࡞ᑊࡌࡾฝⰾ㓕ẍ࡛ࡡཬᚺࡡ┞㐢
࡞ࡗ࠷࡙よᯊࢅ㐅ࡴ࡙࠷ࡾࠊ
1)⏣⏛ࡼ㸯᪝ᮇ㎨ⰹ໩Ꮥఌさ᪠P518(2005)

Comparative analysis of gene expression at heat shock between Aspergillus oryzae and Saccharomyces cerevisiae
Noriko Yamane, Yasunobu Terabayashi, Motoaki Sano*, Kumiko Takase, Shinichi Ohashi*, Masayuki Machida(AIST,*KIST)

P-62
Carbon and nitrogen repression of the expression of citA gene encoding citrate synthase in
Aspergillus nidulans
Pil Jae Maeng* and Yeong Man Yu (Dept. of Microbiology, Chungnam National Univ., Daejeon 305-764, Korea)
The citA gene of Aspergillus nidulans encodes a citrate synthase which catalyzes the condensation reaction between
acetyl-CoA and oxaloacetate yielding citrate, the first step of the TCA cycle. As an extension of the previous studies on
the citrate synthase of Aspergillus nidulans and the citA gene, we studied the regulation of citA expression especially bycarbon and nitrogen sources in the present research. According to the results obtained by real-time RT-PCR analysis,
the expression of citA was strongly induced by acetate. However, the level of citA transcript was decreased by approx.
80%, when either 1% of glucose or sucrose was added to the inducing medium together with 50 mM acetate. This result
suggests that the induction of citA expression by acetate was significantly repressed by either glucose or sucrose. The
extent of the carbon catabolite repression was decreased by about 40% in creAd30 mutant when compared with that ofwild-type strain, indicating that the repression is at least partially CreA-dependent. In addition, the level of citAtranscript was decreased by approx. 40% in the presence of 5 mM ammonia when compared with that in the presence of
5 mM urea. This result suggests that the expression of citA gene is also subject to repression by ammonia.

81

P-63
RNAi 䛱䜎䜑㯑Ⳟ(Aspergillus oryzae)㐿ఎᏄ䜹 䜨䝰䝷䜻䝷䜴⣌䛴㛜Ⓠᒜ⏛㻃 ಞ䟾ᆊᮇ࿰ಆ䟾⚽⏛㻃 ಞ㻃 䟺㒿⥪◂䟻㯑Ⳟ(Aspergillus oryzae)䛵䚮ΰ㒿䜘䛵䛞䜇䛮䛝䛥㔂㏸⏐ᴏ䛱ᗀ䛕ฺ⏕䛛䜒䛬䛊䜑᭯⏕⣊≟Ⳟ䠏䚸ᅗⳞ䚹䛭䛈䜑䚯㎾ᖳ䚮ᙔ◂✪ᡜ䜈ཤ⏤䛝䛥 EST 䛐䜎䛹䜶䝒䝤よᯊ䛒⤂஡䛝䚮㯑Ⳟ䜈䝡䜽䝌䜶䝒䝤◂✪䛴᫤௥䜘㎼䛎䛥䚯㯑Ⳟ䛴᭯⏕ᙟ㈻䜘ฦᏄ䝰䝝䝯䛭⌦よ䝿ᚺ⏕䛟䜑䛥䜇䛱䛵䚮㐿ఎᏄ◒ቪ䛱䜎䜑ᶭ⬗よᯊ䛒ᚪ㡪䛭䛈䜑䛒䚮◒ቪ⏕䜯䜿䝇䝌ᵋ⠇䛴↱㞟䛛䜊┞ྜྷ⤄ᥦຝ⋙䛴఩䛛➴䛒◂✪䛴䝠䝌䝯䝑䝇䜳䛮䛰䛩䛬䛊䜑䚯ୌ᪁䚮ິ∸䝿᳔∸⣵⬂䛰䛯䛭䛵䚮RNAi ⌟㇗䜘ฺ⏕䛝䛥ᵾⓏ
㐿ఎᏄ䛱ᑊ䛟䜑஦ᮇ㙈 RNA 䛴⣵⬂හᑙථ䝿Ⓠ⌟䛱䜎䜑㌹෕ᚃ㐿ఎᏄ䜹䜨䝰䝷䜻䝷䜴⣌䛒಴ื㐿ఎᏄᶭ⬗よᯊ䛴ຝ⋙ⓏᡥἪ䛮䛝䛬Ἰ┘䛛䜒䚮Ὡ⏕䛛䜒䛬䛊䜑䚯䜄䛥䚮⣊≟Ⳟ Neurospora crassa, Magnaporhte oryze, Aspergillus nidulans
ཀྵ䛹 A. fumigatus 䛱䛐䛊䛬䜈 RNAi 䛱䜎䜑㐿ఎᏄ䜹䜨䝰䝷䜻䝷䜴䛒᭩㎾ሒ࿈䛛䜒䛥䚯䛙䛴 RNAi 䛱䜎䜑㐿ఎᏄ䜹䜨䝰䝷䜻䝷䜴⣌䛵䚮᭯ງ䛰䝡䜽䝌䜶䝒䝤◂✪䝈䞀䝯䛮᭿ᙽ䛛䜒䜑䛙䛮䛑䜏䚮㯑Ⳟ䛱䛐䛗䜑᭯ຝᛮ䛱䛪䛊䛬᳠ゞ䛟䜑䛙䛮䛮䛝䛥䚯㻃 㯑Ⳟ䜶䝒䝤 DB 䜘᳠⣬䛝䛥䛮䛙䜓䚮N. crassa 䛱䛐䛊䛬 RNAi 䛱ᚪ㡪䛮䛛䜒䜑 QDE2 䟺argonaute family protein䟻ཀྵ䛹
DCL-2(Dicer)䝟䝦䝱䜴䛴Ꮛᅹ䛒☔ヾ䛛䜒䚮RNAi 䛒㯑Ⳟ䛭䜈ᶭ⬗䛟䜑ྊ⬗ᛮ䛒♟ြ䛛䜒䛥䚯䛣䛙䛭䚮䜷䝷䝕䝅䝚䝯䛰โ㝀㓕⣪䜹䜨䝌䜘ᣚ䛧䚮䛛䜏䛱 Gateway 䜻䜽䝊䝤䛱䜎䜐㯑ⳞⓆ⌟䝝䜳䝃䞀䛾ᑙථྊ⬗䛰䝛䝭䜽䝣䝍 pGFPi ཀྵ䛹 pMtli 䜘షᠺ䛝䚮䛣䛴᭯ຝᛮ䛱䛪䛊䛬᳠ゞ䛝䛥䚯㯑Ⳟ䛴ฦ⏍Ꮔᙟᠺ䜘โᚒ䛟䜑㌹෕ᅄᏄ brlA 㐿ఎᏄ EST 䜳䝱䞀䝷䜘ฺ⏕䛝RNAi 䛱䜎䜑䜹䜨䝰䝷䜻䝷䜴䜘モ䜅䛥䛮䛙䜓䚮α-amylase 㐿ఎᏄ䝛䝱䝦䞀䝃โᚒୖ䛱䛐䛊䛬 RNAi 䜷䝷䜽䝌䝭䜳䝌䜘Ⓠ⌟䛛䛡䛥ሔྙ䛱ฦ⏍Ꮔᙟᠺ䛴㐔䜒䛒ふᐳ䛛䜒䛥䚯䜄䛥䚮䝒䞀䜺䝷よᯊ䜎䜐䚮brlA 㐿ఎᏄ mRNA 䝰䝝䝯䛴΅ᑛཀྵ䛹ฦよ⏐∸䛱⏜ᮮ䛟䜑䛮⩻䛎䜏䜒䜑䜻䜴䝎䝯䛒᳠ฝ䛛䜒䛥䚯䜄䛥䚮㯑Ⳟ RIB40 䜶䝒䝤୯䛱 3 䜷䝘䞀Ꮛᅹ䛟䜑 α-amylase 㐿ఎᏄ䛱ᑊ䛟䜑 RNAi 䛱䛪䛊䛬䜈᳠ゞ䛝䛬䛊䜑䚯
Gene silencing with RNAi in Aspergillus oryzae
Osamu Yamada, Kazutoshi Sakamoto, and Osamu Akita (NRIB)

P-64ࢠ࢙ࣤ㓗⏍⏐⣊≟Ⳟ Aspergillus niger ࡞࠽ࡄࡾ⥫Ⰵ⺧කࢰࣤࣂࢠ㈻ࢅ࣏࣭࣭࢜ ࡛ࡊ࡙⏕࠷ࡒ alternative oxidase 㐿ఎᏄ㸝aox1㸞ࡡⓆ⌟よᯊ
ᑚἑ㈶ྍ㸡᭱㒂㈏⃀, ᮄ㔕㑝ჹ㸡᱊ᮟකኯ㑳 㸝᪡ኬ࣬⌦ᕝ࣬ᚺ໩㸞࠘┘Ⓩ࠙ࢠ࢙ࣤ㓗⏍⏐⣊≟Ⳟ A. niger WU-2223L ࡢ㏳ᖏࡡࢨࢹࢠ࣑ࣞ࿣ྺ⣌ࡡ௙࡞ࢨ࢓ࣤ㟸វུᛮ࿣ྺ⣌ࢅ᭯ࡌࡾࠊᙔル࿣ྺ⣌ࢅᵋᠺࡌࡾ㓕⣪ alternative oxidase㸝AOX㸞ࢅࢦࣛࢲࣜࣃࢺࣞ࢞ࢦ࣑㓗࡞ࡻࡽ㜴ᐐࡊ࡝࠿ࡼ
ᇰ㣬ࡌࡾ࡛ࠉࢠ࢙ࣤ㓗⏍⏐㔖࠿ⴥࡊࡂ఩ୖࡌࡾࡆ࡛ࢅᙔ◂✪ᐄ࡚ࡢ᪺ࡼ࠾࡞ࡊ࡙࠷ࡾ1)ࠊᮇ◂✪࡚ࡢࠉAOX 㐿ఎᏄ㸝aox1㸞࡛⥫Ⰵ⺧කࢰࣤࣂࢠ㈻ EGFP 㐿ఎᏄ㸝egfp㸞ࡡ⼝ྙ㐿ఎᏄࢅᑙථࡊࡒ A. niger ᰬࢅష⿿ࡊࠉ⛸ࠍࡡᇰ㣬᮪௲ୖ࡞࠽ࡄࡾ aox1 ࡡⓆ⌟ࢅ EGFP ࢅ࣏࣭࣭࡛࢜ࡊ࡙よᯊࡊࡒࠊ࠘᪁Ἢ࠽ࡻࡦ⤎ᯕ࠙Aox1 ࡡୖὮ࡞ egfp ࢅ㏻⤎ࡊࡒ⼝ྙ㐿ఎᏄ aox1-egfp ࢅష⿿ࡊࡒࠊࡆࡿࢅ A. niger WU-2223L⏜ᮮࡡ aox1 ◒ቪᰬ㸝DAOX-1㸞࡞ᑙථࡊࠉaox1-egfp ࢅ 1 ࢤࣅ࣭ಕᣚࡌࡾᰬ㸝AOXEGFP-1㸞ࢅష⿿ࡊࡒࠊ
AOXEGFP-1 ࡡ⺧ක㢟᚜㙶ふᐳ࡞ࡻࡽ EGFP ࡡ⥫Ⰵ⺧ක࡛ MitoTracker® Red CMXRos ࡞ࡻࡾ࣐ࢹࢤࣤࢺࣛ࢓
ࡡ㉝Ⰵ⺧කࡡన⨠࠿ྜྷୌ࡚࠵ࡾࡆ࡛࠾ࡼࠉAOX ࡡ࣐ࢹࢤࣤࢺࣛ࢓ᑻᅹᛮࢅ☔ヾࡊࡒࠊࡈࡼ࡞ࠉࢡࣜࢤ࣭ࢪ⃨ᗐࢅ 10 g/l ࠾ࡼ 120 g/l ࡱ࡚ṹ㝭Ⓩ࡞ን໩ࡈࡎ࡙ᇰ㣬ࢅ⾔ࡖࡒሔྙࠉᖏ࡞ EGFP ࡡ⺧ක࠿☔ヾࡈࡿࡒࡆ࡛࠾ࡼࠉ
aox1 ࡢࢡࣜࢤ࣭ࢪ⃨ᗐ࡞౪Ꮛࡎࡍ࡞Ⓠ⌟ࡌࡾࡆ࡛࠿♟ြࡈࡿࡒࠊ
1) K. Kirimura et al., Biosci. Biotechnol. Biochem., 64, 2034-2039 (2000)
Expression Analysis of the Alternative Oxidase Gene (aox1) with an Enhanced Green Fluorescent Protein as aMarker in Aspergillus niger Producing Citric Acid
Satoshi Ogawa, Takasumi Hattori, Kuniki Kino, Kohtaro Kirimura(Dept. Appl. Chem., Sch. Sci. Eng., Waseda Univ.)

82

ெྞ⣬ᘤ
஬ᒜ┷⌾··················································· 32ᒷᓧ㜷ᑋ⨶ ··············································· 53
ᒷୖ࿰⿩··················································· 19Ꮻఫ⨶ㄵ··················································· 55
⁵ཋᑋᙢ··················································· 51
ᾇ⩹ሪ㇇········································75, 76, 77
ኯ⏛᪺ᚠ············································· 48, 63ኬ᰷⏛Ꮼ··················································· 49
ኬ⟺ಘୌ············································· 40, 80
ᒱᓧᏏ᫆··················································· 65ᑚ➗ཋ ༡ಘ ··········································· 42
ᒱ⏛᫥౅··················································· 57
ᒱ㒂᪺Ꮔ··················································· 50
ᒱᮇ⥜Ꮔ··················································· 62
ᒱᮇಆ㍔··················································· 54
ᑚἑ㈶ྍ··················································· 81⧂⏛ ೸··················································· 19
ⴘྙ์ᖶ··················································· 57ᑚ⏷ ᾀ··················································· 42ばἡᨳ஦୔ ··············································· 68
みㆺ├ᶖ··················································· 41➗ཋ ⣸··················································· 44
ຊ἖ఫᖲ············································· 69, 70
᯵ᮄ ㇇·································· 40, 64, 65, 79
ຊ⸠ࣀࢻ··················································· 32
ຊ⸠㞖ኃ·································· 47, 58, 77, 78
㔘᲻ḿᶖ············································· 32, 54
㔘Ꮔ ຉ··················································· 32㙂ಲ㧏ᚷ············································· 53, 54♼ኯ㑳······················································· 62
ஞ஬ගᙢ··················································· 57
ἑහᙹ඼··················································· 59
ᕖᓃ⨶౅··················································· 66
ᕖ㒂┶Ⓡ··················································· 50
ᕖᮟ┷ᚷ··················································· 59
ᕖᮇ㐅······················································· 57
ⳝờ㡝ᙢ··················································· 66
ⳝ㛣㝧ᚷ············································· 49, 75
໪ᮇ์⾔··················································· 64໪ᮇົࡥࡆ ······················· 36, 41, 42, 48, 49,····························60, 61, 71, 72, 73, 74, 75
ᮄ㔕㑝ჹ·············································· 43, 81
ᮄୖᾀ······················································· 45
ᮄᮟኣỜ········································40, 64, 65

Drivinya Antra··········································· 57
David B. Archer························ 29, 36, 61, 64
Mee-Jeong Cha ·········································· 66
Suhn-Kee Chae ·········································· 66
Paul S. Dye················································ 61
Evelyn B. Elegado ······························· 52, 53
Dong-Min Han··········································· 60
Kap-Hoon Han····································· 35, 60
Howard S. Judelson ··································· 44
Praveen R. Juvvadi ············· 36, 48, 61, 73, 75
Hyo-Jung Kim ··········································· 60
Jee Hyun Kim ············································ 60
H. C. Kistler··············································· 50
Nak-Jung Kwon ········································· 66
Heather A. Lee··········································· 64
Pil Jae Maeng ············································ 80
Maria Dolores Montiel······························· 64
Sun-Hee Noh ············································· 66
Berl R. Oakley ··········································· 66
Richard O'Connell ····································· 52
Mathieu Paoletti········································· 61
Nguyen Bao Quoc······································ 41
Fabian A. Seymour ···································· 61
Marites A. Sales········································· 53
Sung-Hwan Yun ········································ 34
Yeong Man Yu ·········································· 80ྙ⏛⚵▦··················································· 78⚽⏛ ಞ··················································· 81㜷㒂 Ṅ············································· 52, 53
㜷㒂ᩏᝃ········································38, 47, 63
Ꮽ㒂ᗛ஁··················································· 79᭯Ờ ງ·································· 32, 50, 51, 54
᭯ᒱ Ꮥ································· 11, 41, 42, 49,································· 60, 74, 72, 73, 74, 75
᭯Ἁ ❮············································· 45, 68஫༎ᔋᅽ᪝Ꮔ ································68, 69, 70⏍㥎༜ஒ··················································· 50▴஬⤦⨶··················································· 76▴஬༐὘··················································· 53▴ᕖᬰᏄ··················································· 50▴⏛ ༡ᶖ ··············································· 25▴⏛༜ஒ··················································· 69
ᕰᕖீኯ··················································· 56
ୌᓞ ⱝ἖ ················································· 46
ୌᰏಆ௒··················································· 63ୌ▴᫓ᙢ··················································· 59
௿⸠ᖶᠺ··················································· 71

83ᮄᮟ┷················································· 56, 57
᱊ᮟකኯ㑳··········································· 43, 81㔘㗋᮵······················································· 73
ࢡ࢙ࣤ࣬ࢤࣤ࣬ࣀ ··································· 62
ᴃᮇ៿ୌ·································· 40, 64, 65, 79
ᕝ⸠យᏄ··················································· 59ᕝ⸠ಆ❮··················································· 55
஁ಕᗛ஄··················································· 52ಲ㒌ᑑᏳ··················································· 72ᖶ⏛ ᪺⏍ ··············································· 23
ᑚᓞᾇᖲ··················································· 58ඡ⋚ᇱୌ㑳 ··············································· 50ᑚᯐலᕵᏄ ··············································· 40
ᑚᯐူኰ·································· 47, 58, 77, 78
஫࿝ົஒ·································· 21, 42, 47, 63㰳⸠៿ୌ㑳 ··············································· 54
᩟⸠⚵ᠺ··················································· 51
㒿஬Ᏻ······················································· 51ᆊ㔕┷ᖲ··················································· 56
ᆊᮇ࿰ಆ············································· 17, 81
ᆊᮇ⿩ୌ··················································· 67▹ᕖ⨶ఫ⥬ ··············································· 71ྡྷᆊ⏜㈏··················································· 55➪ᰡᚷಕ··················································· 74
ఫ⸠ฺḗ··················································· 67ఫ⸠ཪ⣎··················································· 71
ఫ㔕ඔ᫓········································40, 47, 80
ఫ㔕⿩ᇱ··················································· 76㩢ᓞḿᾀ········································68, 69, 70
᳕ྞᮿᏄ··················································· 47
ሲ⃕࠵ࡍࡈ ········································· 56, 57
ΰỀපᚠ··················································· 57
ΰỀ೸㞕··················································· 45
ᚷỀඔ஼············································· 45, 46
ḿ㊪ῗஒ········································41, 42, 72Ⓣ▴ཪ⣎··················································· 52ᮙᒜ⣟ஒ··················································· 78
㕝ᮄᚥ······················································· 50㕝ᮄ ⪵············································· 65, 79㕝ᮄୌྍ··················································· 68㕝ᮄ⪵················································· 40, 64
㡪㈙ᬍ஁··················································· 50
ເࠎᗛ௥··················································· 75
᭧᰷㍜㞕············································· 52, 53⏛὾⥜Ꮔ··················································· 72㧏℡஁⨶Ꮔ ··············································· 80

㧏㔕⩇Ꮟ··················································· 58㧏ᶣ ▹⧂ ··············································· 42㧏ᶣ᪺⌌··················································· 78➁හ㐠㞕············································· 61, 62➁ୖ඼⏠··················································· 48➁ㆺ༡Ꮔ··················································· 65ኣ⏛⨮ Ὂኯ ············································· 46㎦ᕦᬏ⣎··················································· 75⏛୯༐ᑖ············································· 57, 58⏛୯ᑋᇱ··················································· 78
ㆺಞ἖······················································· 44⋚㔕Ꮟୌ··················································· 71
ሪ⏛๙ኃ··················································· 70㎱ ඔெ····················································· 52㎱ᒜ ᙪୌ················································· 52
ላᾀᏄ······················································· 79ᜇᕖ ༡············································· 45, 68ᑈ஬᪺✉··················································· 57
ᑈᒱ ᚥ·································· 32, 50, 51, 54
ᑈᯐ㟱ᐃ············································· 71, 80
ᑈᮇ ᐰ··················································· 46
ᐣᶌຊዄ············································· 32, 51
ᚠᒱ᪸ᩝ··················································· 63Ễ஬ྍ㑳··················································· 45
୯ฝၤᏄ··················································· 67
Ễ஬ົ······················································· 67
୯ᓞబ······················································· 47
୯℀ᬓ஄········································42, 60, 74
୯ᮟ㈏······················································· 77
୯ᮟ▩ᜠ··················································· 68
୯ᮟ㞖သ··················································· 67
୯ᒁᩔᆍ··················································· 41㞬ἴ ๙····················································· 46
こᮟ㯖㔓Ờ ··············································· 43
஦༎㌲᜾㔓 ··············································· 70
ொᖲ༜ஒ··················································· 45୸⩒ಞ㌗··················································· 65㔕ཾ♰ྒྷ··················································· 78㔕ཾⲸᯖᏄ ··············································· 56
ⴏཋኬ♰··················································· 47
␂ᒜ⌦ᗀ··················································· 60⛑Ὂ஦················································· 42, 79
᭱㒂㈏⃀·············································· 43, 81㤷ሔᓧົᙢ ··············································· 67ᯐ㛏⏍······················································· 43ᵵཾ⿩ḗ㑳 ········································· 42, 6084஁⏛༡ඔ··················································· 79Ⓤ⏣‫‮‬᭹··················································· 50᪝ྚ࠵ࡷᏄ ··············································· 69ᖲ▴ḿ⏠··················································· 69
ᖲሪᐃ༡··················································· 68
ᗽᮆួႵ············································· 45, 68
⸠஬ ໌········································75, 76, 77
⸠ᒱᬓ์··················································· 63⸠ᮟ┷············································51, 56, 57
ఄಘ㐅▦··················································· 70
ཿᕖ⫩௥··················································· 40ཿᕖ೸ኯ㑳 ········································· 38, 63⣒ ᭰ᾀ··················································· 66
ᇷහ⿩஄········································15, 48, 63
ᇷᑹူஒ··················································· 66ᇷ㉲Ờ⨶··················································· 71∶⏛ᬓ⿩··················································· 77⏣⏛㞖஄·································· 40, 47, 71, 80
ᮿ஬⌦ᜠ··················································· 52ᮿᑹୌ㑳··················································· 71ᮿᓧⰟ⨶Ꮔ ··············································· 45ᮿୖ┷⏜⨶ ····································40, 65, 79
ᮿᯐⰃ༡··················································· 58
㤷ᶣ⏜౅··················································· 74┶ᒜ⁘ᚷ··················································· 41୷஬ῗୌ᭹ ········································· 47, 58
୷ᒜ₮ୌ·················13, 36, 48, 61, 71, 73, 74
Ềㆺ἖················································· 47, 63Ề㔕⊓······················································· 47
୔⏛ᆀ㈏ྍ ··············································· 54୔▴ Ꮽ··················································· 69
ᐋᓧᏭᑑ··················································· 67
ᐋᆀಆᙢ··················································· 52Ṃᕖ ἖··················································· 53ᮟᒜ⫍Ꮔ··················································· 59
Ⱪἠᖶኯ··················································· 59
ᮇᒜ㧏ᖶ··················································· 55
᲻ཾ ᬓ⨶ ··············································· 77
᲻ἃ⧂······················································· 51
᲻⏛ᐰெ··················································· 61᲻⏛┷⣟··················································· 55▦㏛ග㑳··················································· 69
ᒜᙟὊᖲ··················································· 47
ᒜཾຩ······················································· 56
ᒜᓧᬍ୓··················································· 63ᒜᓧࡹ࠾ࡽ ··············································· 58ᒜ⏛ ಞ··················································· 81ᒜ⏛ಞྒྷ··················································· 68
ᒜ᰷೒Ꮔ············································· 71, 80
ᒜᮇ ୏℡ ··············································· 73ྚ⏛ Ꮟ····················································· 46
ྚ⏛ㄌ······················································· 70
ྚぜၤ················································· 57, 58
࿰⏛᭰Ꮔ··················································· 70ῳ㑌⟿Ꮔ··················································· 56
ῳ㑌๙ᚷ··················································· 47
ῳ㎰ူ······················································· 57
ῳ㎰ὀ♰··················································· 73
๪▴༡஄········································45, 46, 68

85

糸状菌分子生物学研 究会

会則

本会を糸状菌分子生物学研究会（Fungal Molecular Biology Society of Japan）と呼ぶ。また本会が開く研1.

究会を糸状菌分子生物学コンファレンス（Conference on Fungal Genetics and Molecular Biology）と呼ぶ。
2. 本会は糸状菌の分子生物学、細胞生物学、生化学、生理学、遺伝学などの普及発展を目的とする。
3. 本会はその目的を達成するために次の事業を行う。
(1)

研究会及び総会の開催。

(2)

会報の発行。

(3)

関連研究団体との協力事業。

(4)

その他、必要と思われる事業。

4. 本会はその目的に賛同して入会した個人会員及び総会において承認された名誉会員を持って構成する。
5. 本会入会希望者は所定の入会申込書を提出し、別に定める入会金を納入するものとする。
6. 本会はその運営のため、会長、運営委員若干名および会計監査 1〜2 名をおく。任期は 2 年とし、改選は運営委員の推薦と総会の承認による。
(1)

会長は本会を代表し、会務を統括する。

(2)

運営委員は運営委員会を構成し会務を審議する。運営委員には庶務、会計、編集担当、広報担当をおく。
会計監査は本会の会計を監査する。

(3)

7. 本会は事業運営に必要な実費を年会費として個人会員から徴収する。
8. 本会の事務年度は研究会の開催準備開始から「次期」研究会の開催準備開始直前までとする。
9. 前事務年度の庶務、会計については、これを総会において報告し、承認を得るものとする。
10.

本会則の改定には総会出席者過半数の賛成を必要とする。
以上

補則
(1)

本会則は 2001 年 7 月 1 日より発効する。

(2)

本会入会金は 1,000 円とする。

(3)

年会費は一般会員 2,000 円、学生会員 1,000 円とする。

(4)

研究会の通知及び会報は、当該年度までの会費を納入した会員に送付する。

(5)

2 年度にわたって会費納入のない会員は、その資格を失うものとする。

(6)

研究会の発表者は、会員に限るものとする。新入会員の演題申し込みは会費納入の確認を持って受理する。

86

糸状菌分子生物学研 究会運営委員 会名簿
会

長

北本

勝ひこ

東京大学大学院農学生命科学研究科（〒113-8657

東京都文京区弥生 1-1-1）

運営委員
秋田

修

独立行政法人酒類総合研究所（〒739-0046

五味勝也

東北大学大学院農学研究科（〒981-8555

鮫島

広島県東広島市鏡山 3-7-1）

仙台市青葉区堤通雨宮町 1-1）

正浩

東京大学大学院農学生命科学研究科（〒113-8657

東京都文京区弥生 1-1-1）

会計担当
有江

力

東京農工大学農学部（〒183-8509

竹内

東京都府中市幸町 3-5-8）

道雄

東京農工大学農学部（〒183-8509

東京都府中市幸町 3-5-8）

編集担当
小林

哲夫

名古屋大学大学院生命農学研究科（〒464-8601

名古屋市千種区不老町）

広報担当
川口

剛司

大阪府立大学大学院農学生命科学研究科（〒599-8531

大阪府堺市学園町 1-1）

庶務担当
堀内

裕之

東京大学大学院農学生命科学研究科（〒113-8657

東京都文京区弥生 1-1-1）

87