Division of Biofunctional Chemistry
The Chemical Society of Japan
Vol. 20, No.3 (2005. 12. 8)

   g \   lDNW







[o` stxB>54+.Km8,;</0*=3;eDMl
diHU |py'),9AqL]NWq^mZcEB
 Tw>%<#=gG6)-Km           1"/08,;<M~hnXI bDmfz_JVevZOj
 C}@ u  RlDfzNW'=7(9

FSP

 u  RlDfzNWCk
 {aC&8?)$?<
 {a2!?:9

FSP

FSP r
 uRlQNWstC'=7( 

9YZ

   









 

ōŋŋŐÇőè5©¤Ã1ąòĞłĵ½Ļ6ōŋ«ÇÐUsgVNj!ĦHF-²óĮ¯ą!,6ı6ļēńñòĞI?/éò½¢Ï6čđ6 ŋÇŁIª>0ŎŋÇŁ6ô¥IÙDĳ.0ĀĬýI(F-ġ¡Ě5ÔÓIě#2>0ě-'2
1D,J4ô¥!.-6
t^)0

2ņ

FÊ54.-26ÔÓ?¶.-čđ|ąIWYC'6ľ1Ĝ.0"-ĤĚ52.0?Ü>0GG2ęEò54.-ĤĚ7¨Î·½·½Ń6µīďIî)0*$5zÅ·½6²óčđÀ6Ö54.

-µīďå~57ķĽKrMp6oVPp¢Ï!čđ\th1Dz·5İ)-2"5²óąCļēl]p¢ÏI*E

C2ĨHF-éòü¹x6¢Ï

4Dļē6'243?ČC4ı52.07¸³­uĺą!æ
=!Ŀæ1.-    Ĥ      Iuĩ=ĕ024#ļē¢Ï!HC÷üúx6¢Ï5F-ļē¢Ï6)#     EB4ö!)0"-Ëå6ļēńñò

ĞI?/éò½¢Ï6čđ7ä6Ç6á5DĪBD?â#ÖIØ&424

4

®1"4;3þö9FEčđ1.-<Ñ8,6Ëå7÷üúx1Ħéò¢ÏIĦ.0-61.0ąňļēŉ¢Ï57;3ĸāÕ1.-

ì{Æî·½5ĎDĀ5ŌŎÇ5ą¬ĄÆ½Ļ5Ď.0

C7Đïċ5bMOľ5yD)}17čđÀ6·!ĹºĖ=Ú@·ğĢ@ÿĔĝI.-YscR@ļē
6Ċă5£DĖJ1EĠ6½J1"-éò½5õ:0ą½@ºąÿ½7B#h`m
Kp(Fĥ43?Ň17E!Q[^(FktPt?
4Tgt^

A2'G5Ö!Â#B

!ß.0Eéò½?ą½6¿Ņ?´êċ57  6-<D1

DĘ2'G7¶ą½ł6½17ĥktPt@û¾ò±ktPt?osZnsXi_t43Iŀ)0Đïċ5½5¡)0E'6?Ę-ą½17íĽ
7   6ŏà»1KiaĽ?uà»ħ1D½ðĴÉIæ

4#4.0E))'67Ę-#74ą½6Ĭė1?4E:#½ðĴÉIæ04+,46

Iºqfp1ÝEB5Í!%0EËĻ7ŎŋÇ5ą

1'E¢ÏIéò½6Ĩģ1Ą*E'2

CWYt^)-çĲ7ĻUsgVNj6Ċħ6w7ą?6I×B54.0Eá6ä
¼Ç6ĊħÞ6x1?ŌŊŎ!ą

łĵ1Du·1Eáê½57ąòĞłĵ½ĻbMO\RarVtĻą¬½čđ6v/6ą

łĵ6Spte!D¦!ĂĒ)0UsgVNj43IĶ°)0Eáê½!]LdVnsI£
DFE'2?DÁëċ57§¨E7ĵÛ)0éò½6ČĭIùć)//ą¬ăįIºqfp1ã)ĆîĈ2?ĵÛ)4!CĥÈĉÒ6ø4365ÌĒ0Eľ2)0ĊÄ*Eñ5Í!%-

*,-10.&2453) 

 

"



'+

$/ (*,! #%       
1.+2)
Äºg³º<;PR=HfWb¹ïs*©gÆ¦*ÏÔ+ăÈ*PRLBReEh)
%²3k¤<(3*&8'+ê0&3(ĄPRLBReEh*¿+ăÅ<PRD>G)0&¢Û8'&ăTcBÇ¨'+Ì(8Æ³*Å¦ÍÈ´¡98':60#%
8ăÈ&+ăPRBaFIhÑýgy¥Ö¦Õ<®ÀÊ8PR`QKN=HfWb*
ÏÔ)Ó7$$7ă

*k&ăĂ|*­$åÒä<Ê!PR\Lb=c=HfWb*ø

Íç;9%8Ą &măåÒä<­$Ă|'%T>@|&8 DNA 4YXJO<Ê%ªç#%!ăPR`QKN*®4=HfWbÏÔ<Ün8Ą
2. DNA Ă»¹ï<~Ê!PRØ=HfWb [1]AuSí Mao 6)5#%ăÉs{*òq¯*û)1698
ZbM>ðÞÞ<~Ê!ăÙ 10nm *r¸ô}!ß¹ï*¹×9%8Ą*¹ï+ă

Au

Au
Au
Au

9!ô}<­$ 8 ¶( 6 ¶* 1 ¶ö DNA ă4$*ZbM>ðÞÞ<)!PRr¸<©ă6)µó|)¼%8öó|<~Ê%ü®8o*r¸' 2 »xÎ)Þ8')5#%©98Ą

ª+ă* DNA Ă»¹ïs<PRMT>F*LfXdh
N'%'6ă

Au

Au
Au

Au
Au

Au
Au

Au

(a)

(b)

 1. DNA PRr¸<~Ê!õP
RØ=HfWb*i)PRØ<Lhah^>O)=H

fWb&8'ã!( 1)Ą0 Mao 6
Rr¸ù*ðÞ)u;98ą±

! DNA P*ą¶öIC<õPRØ)%8 DNA 'éÎ(ô})â¯8Ą
%ăDNA vāõPRØ'z)Ă»¹ï<©8'ă

õPRØ 1 »ô}8'l§98ĄûăýĀ¢÷&Â!':ăõPRØ=HfWb9%8'Ðë9( 2)ă¶«¾*³¦<!Ą

 2.  1 )57û)=HfWb9!õPRØ* TEM Ëw

3. DNA åÝá<~Ê!»jp½Ú¥Ö¦PRS>WbKO·°[2]DNA *Æ£'%ăþ)ñ¬¦*ĂéÎ(Þ*o)ăĂ|ýìýæ<.![b\h&7ă*Æ¦<­$ĄDNA +*èÿ)õ>@f<Ãà8¦8Ą6)ăDNA +î-3Ç"ăõ>@fÃ8')57CeV_hc¹ï',98Ái¹ï<©8Ą

ª+ă* DNA *AJ@f>@fÃàºä'CeV_hcÓ<~Ê%ăõPRØ<=Hf
Wb&8'ãă½ÚHfDh/*¥Ê'%¿9%8UaE?](Pd))ú%ăDNA-Pd
S>WbKNPR=HfWbs*t©<1!Ą

-Ć-

Pd

¦À,iÇ)&(èá¥V¹Æ), 

ª

se),

(è~q¦À8I4K

Q³Êµ*)é¦ÀiÇ&)á(a) ¦ÀiÇ&)á¥se,b°
 Pd ¦À8I4K
U, ¦ÀTv17äw¥se,b°@E6/C5K;¦À8I4K(l3a)Ü

Ú²

Ò$'*)è\§*)á§Ø¤Íâ~á¥¼,Y*!'$èæ¦ÀTv17ÀÔ

ãËå

i´%á¥ã%¡)é"è

uf)$è¦)é

 Pd ,_èPd =>¾xèDNA M)LzÛÝ,ºÖa)ÐV,W)&è¦ÀiÇ&)V¹Æ Pd =>¾xht

#á§§*)57:C½, (l 3b)é(b) ¦ÀiÇ&)V¹Æ,b°
Pd ¦À8I4K
æ, ¦ÀTv17ä©

 l 4 g Pd  TEM ±[,¶é-DNA âyo
N

è-73GAI×Õ\^®

'*(a)èNaBH4 Õ\¿ 200nm ¾`¾x
¾(b)é*

p¾xâ~ Pd ¾xRß]ÞV

'*

è-73GAI× NaBH4 Õ\cÓ²¯ÏÙÏä,O
-DNA yoNÁ)éLè

è-73GAI×Õ\rgè¿ 5nm ¾xÞg)¿ 70 nm ¾`¬æ` Pd ¾x

'*(c)èNaBH4 Õ\%è)¸pL¾x(p¾¿ 25 nm)Ñ)«l 3. Pd ¦À8I4K|

*(d)é&èDNA  Pd .0I,¨ÃÕ\)è¾`~ Pd ¾x*)Éè'èDNA 2HADKGÐ·Õ\ÏÓ&(èVÐ%uf)

)

¶j*)é ¢è-73GAI×Õ\ Pd =>¾x,áÛmzán,êíLzZè5 `£»ÀJ1ç¦Ààk¥Nufá§uf,ÎÌ(l 5)é
DNA âyoNÕ\ Pd ¾xè¦À¼,¶}è-73GAI×Õ\W DNA-Pdè¦À,iÇ)&V¹Æ

9<VèDNA-Pd ?.BF

¿ 5 nm  Pd ¾xÞgV)

iÇ&( 5 nm  Pd ¾x)$Ä

'è¦À

V¹Æã¹,se)é

TMè*"èDNA yoN Pd .0I,Õ\)&èDNA 2HADKGÐ·,b°
Pd =>ÐV,Èè¢PSq Pd ¦À8I5I2Å,²­)+"d)é

160

5.5

N2 H2

140

N2

H2 N 2H2

N2

Current ( -DNA)/ mA

Xyqá¥¼,¶é*Current (+DNA)/ nA?.BF9<V

l 4. Pd ¾x TEM ±[(a) -73GAI×Õ\ Pd, (b) NaBH4 Õ\ Pd, (c) -73GAI×Õ\ DNA-Pd,(d) NaBH4 Õ\ DNA-Pd

5.0
4.5

120

4.0

100

3.5

80

3.0

60

2.5

40

2.0

20

1.5

0
0

300

600

1.0
900 1200 1500 1800 2100
Time / sl 5. Pd ¾xá¥¼ÂìPd ¾x, {ÂìDNA-Pd ?.BF9<V-ë-4. N58WK[\Bb>Z`0~ÝVF\3]ILÃĀo
üđ(=`<!hĆÕê0+

1óþ

p©[3]

ēÀ

,ÕĆ9]>6X0©ĕgđ'9]>6X58`*ē(È,\`Ć9]>6X0©,ė$ĕÛö

!ĕó{óþËď¶ÛĆ0ÝďéñÅÄ0%ĕIL^T]
A`R\0ø,ė


58`

IL:[@Cb3

N58WK[\Bb>Z`!ĕ*WK[]0ĉ,§¢ĕÇâÖÆî¶©ċ0£ĕIL

ĕĄn
*A`D

^T]$ôĂēÀÃĀ0Ãç,ė$ĕÐaf¦ÌÒÏďä(»m¤(ÂČ»m·Æº³ý,&ĕØ  6. USDH0~ÝVF\3]ILÃ

M5R\EH3A`R\Đ Āo p©

'ĕN58WK[\Bb>Z`!Ĕà,ėÿĕÖÆî¶îUSDH!ĕN58WK[\Bb>Z`Æò0·,','-,ė

ª!-*ÖÆî¶îa©USDH0ÝĕÐaf¦ÒÏďä(ÆIL:[@Cb0½d#OCbJ`;+ĕl«Í!ā©ÃĀo0p©,0®,( 6)ė!ĕª»mÖĎIL3A`R\

øĆiĈè

N58

WK[\Bb>Z`ìk,ė
$}&ĕ12 É

USDH0¯ãQ2b?[5R[\b0ÝĕĆiĈĒæwă¬0øė3 ă¬ĕvQ2b?²,ĆiĈîč¦Q2b?²¥Ó 300 sėĕ-*ĆiĈîč¦Q2b?
.ĕ50%


{

Q2b?

cą|0¯ã+ĕ*¯ãUSDHą|0¾

USDH!ĆiĈÚßàî,

ė

ĕ

ĆiĈîč¦USDH0½d

(a)

(b)

(c)

(d)

#,)ĕÐaf¦ÌÒÏfĆiĈ%ă¬à0%ėUSDH

!ĕĆiĈîč¦USDH¹D8b]0tĊ­>@F5`0vĕćĖD8b]î0~ÝćùĐėĕ¸yÜćS^bGUSDHćS^bG0÷u¦ĆiĈè

{±ÌÒÏf30 {Îĕ254nm

ëu0×ĆiĈ0Ázėî¿ĕUSDHS^bG

iĈÞ¼

ïõ÷u

#,©,
$ĕl
SDH

%Ć

Ñ-ĕĆiĈ0½d

ÖÆº³#

/

,( 7)

î'r,&ĕU-,ćS^bG0ĕ÷uÚ¦0­áiĈĆY6_P6Xè¨ÔÏÎ.ĕćS^bGd0Ñ,





èÞ¼

÷u

ĕjă¬-ĆiĈîUSDH ZnOí©0Úßàúû,

ã-ė-!ĕUSDH05`:?7EGåOCbJ`;-"Ðaf¦Ì-Ę-

 7. ćĆÙ¨ÔÏÎ-ćS^bGx
(dÊęëð¸×, f´ę254nm ×, eÊę254nm ×(° µëð9EGQ4]CbqÝ))(a) ZnO ¨ÔÏÎ¡ USDH¸ćS^bG(b) ZnO ¨ÔÏÎ¡ USDHćS^bG(c) ZnS ¨ÔÏÎ¡ USDHćS^bG(d) Eu2O3 ¨ÔÏÎ¡ USDHćS^bG

ÎÉµZPý"ąoTĆç)@6N<L/&)â|%ĖVÒ"¹ċ×¯¾·¯k@6N<L/£ô±&ė
?-.E=HI4N0GLĖÂąoT

Ć5JÕĖ029-L²XQąoTĆëvčDC7:)Ëm&

#



ėËm$5JýÃĊĖ1 «D
C7:)Ëm!Á5JÃĊÏì[äqeYP%Ė3 «êĀ~ăçÕąoTĆÃÀĆ)v&ąoTĆvÍ'ĖÌąoTl( 8)ė!ĖDC7:)ºą)Ëm!û

&,E>$ė,E>ą

ëvDC7:,E>ąĄfR
gąoTĆv

oTĆv

ÃĊÏì!

¾î

ĖąoTĆ#Ė

Ø'ĖÌ×

'&ė$ñ{ÊÔÕ)đĔć#%

ą

ĖÃÀ
(Ėè 20nm ąoTĆ;>çiÞAHKNF7NA)

áõ'( 9)ė 'Ė

&



?-.E=HI4N0GL#%ºþ[)\VDC7:MóÝ
&&

 8. ąoTĆëvDC7:ËmÂąo
TĆ5Jê­o'z')â|%Ė

\%dºþÿë

#%Ė#%ĐïĂºþuî

 9. ąoTĆëvDC7:#%
oTĆÃÀÔ SEM Û^

ą

(ė

øü VéWàåĖ¶p_Ôãàå¡ĉàå268A(ĉĎ¨§¤, x}æn§¤, t®n¢)Øß»(ÜíÄy, ÂÙāy, c½ú
!

'!%,y, ¼°*)rkøðėĖA+N1¦âÆĖ¶pČàåãÑùÅ§¤Ç³Èn§¤(Ö¶UČªējãn§¤)ĖÐ¾´©¦]MawàåĖĈ¥vãàå¡bòÓÚ§¤ĉăÈSn§¤
ė

Ê

rk)Ē

øðė

1. Ü íÄ, s ¬, ¸Ç `, t ®, ĕ öO, ĉ Ď¨, DNA ĕ¿ºþ[h;>ç,3LBI, ĕe÷¨ď, 61, 617-622 (2004).
2. M. Umetsu, Y. Hatakeyama, S. Ohara, S. Takami, T. Naka and T. Adschiri, in preparation.
3. M. Umetsu, M. Mizuta, K. Tsumoto, S. Ohara, S. Takami, H. Watanabe, I. Kumagai, and T. Adschiri,“Bioassisted room-temperature immobilization and mineralization of Zinc Oxide - High order structuration of
ZnO nanoparticles to a flower-type morphology-“, Advanced Materials, 17, 2571-2575 (2005).

-Ę-357 B> </ C;18

2 #+ $(2,0


?&(*(354 ).
       
sjUznFÏ2&+ĠćĜQSYn7mRNA A6Ĵ¸Ĉ4Ġć6Ķ§2
TxZrt[Ml6Ķ4ÊFÜ.0Cİ
Uzn6ŋęėåòĭîA
.06ĐĈ4ĉč95īÅ¼Ā%D+sj

Ŀ4ĠćĜQSYn5¾'C·ugt16ā¤Ğ54

+ĳ·ĴŇĳ·æ¥ňAĥ°­¾ÉÕ2½4

ĦĨõòFă&0

heW[ė§Č¢ÑFÍ&Ĵy·ķ
y+-6½5Í%D+ņ·Ňkshe
W[ňFĶ§'CĐÂ4Ė>7Ņ!<?61C0çī17 mRNA 2sjUzn6

ăóÇ5@.0ĠćĜ6äĻĈÍĢED0C$2?ãA54.+ŀÃ5



Ŀ4$D

A6¢Ñ?ÎĈ5>D8ē4·ĦĨ¹6äĻĈyĎĻĈ4Ė>§E)1BĬÆĈ4
NmSZszy_HJYL^vRz6įö7ĠćĜQSYn6ā523=A(ĈÍyÁ¹ĈŇ¹ĈňÑăF?¤Ğ5&//CÖ7đĸFă&+·ľ§6òď5ļ'Cĉč 1)5īÅĠćĜQSYn6Í2¹Ĉă5¨"+ó4GevzWFĢ.0C|5*DA6ĉč6{5/0èõ¹yìß¹ĈAĕ%)0+,

+




-

"%:

@

A 

69!='2)

O^nQzLIxS6[rcZîŇŉ°­ň´¬ļĮî%A57ĘĚĈĆĀîÚģ6
įÀ5ĳ·ñyàAÌADCÓ®ķ?ŃĪĈ5±µ&0C&&Ëí6ĳ·àyĆĀp\VsxM7'B/:&+Ĕĝ@BÛ&+ DNA/RNA Fă0ĢED0Bĳ·
6³ąĆĀÿø7Ĕĝ6Ä«2&0£BÙED0+ĔĝěF{Ĕĝ6ľ§ŇQSYnň

2+¯§*6QSYn5"C¦{Ĕĝ6ÿÔıFäĻyĎĻĈ`rozV25ý&Í'CÚģ7
QSYn2&06Ĕĝ]XZwzL5¾'Cá+4GevzW24BC$6
@4Ĺé

ô6¼Ā57Ĕĝ6´óÒ6ð1CĔĝğFĊ²&4ĂĔĝĲ§¬6ĳ·ñyàØùÐŁ1CÖ7ĔĝĠćĜQSYnFă&+GevzWŀń RNA F
ăCĔĝĳ·TxQxM5Ý×&0CĔĝğ6@4ü§Ē5"Cĳ·ñyàF¤Ğ5'Cevzd6

Ĥ2&0puKqrzaz

PxMBÞ#ADCMB 7ġyû·F~ê
5è'CfGbxòĭïĵ1BôĈïĵ6iSZz
OSZĈė§5ħĩ%DCòĭ³5@BġFĆ'C&&ġû¬ MB 7÷Òyº»Ò43Ĕĝ26
Ĳ§Ò¡9´Ĕĝ;6¿ù5©łB*6
ă57{»6Ľ.+{âïĂþĄí6 RpoSmRNA 7SYntze¬ŇQSă¬ň6 mRNA òĭ
Fè&ZrxSă¬ DsrA RNA 5@.0Èĩ$%

DC Open-Close ¬òĭ³ŇĠćĜĺ¶5ÐŁ4
sjUznė§Ĵ6úÒ}úÒň5@.0Ġć6ĜFÍ&0CŇª 1aň=+īÅ Collins A5@.0ëõòïĔĝ16ĳ·ĆĀÍ5Ñă¤Ğ1C$2ċ%D0C 3)$DA6ïĵ

ă7

-Ŋ-

ė# MB ÞĘ"ĭ¶"įu/Æ$ÛĂĈďòĆĝs§ÜeÑĀ#Þþ"MB  mRNA(MB-mRNA)#@AH[3ï

ĂĈĚ

3Đàóĝs§ĭ¨i$`WDf[ĄĠRBS: Ribosome Binding Site,Ĕč3 MB T4QdÞĘ"jæÁàóĝs§¨i"#)ďòñíĦ¥0/ĴĶ; àóĝs§ĄĞv, ĳčĵbXfEfďò¦æÁ3Õ/a@R6_fC3ĜÉa@R6`d3

âī#

ġā

¦ó@:La·3)HIV Ĩęĝs§

/;\85d CCR5 OScE5S3àó

 MB-mRNA 3I?5dĴ 1bĵ¢ċĎďòÅ@AH[k"/ĝs§Ü¬ı3Đ

+ amol~fmol bUa#ÜÃ¹àóĝs§3Ü/1³!

(ØĀ  ēĉ3Õ

/öđ0 2a)!in vitro Ā'#¿ï"Ī/Ü+Ó-Ã¹eĜÉÁh"RNase H Activity-Coupled ¢ċĎĆķģ

RNase H æÁ",.MB-mRNA 33ï

Ā3ï

¦@AH[3yÅMB-mRNA(for -galactosidase)3ï

One-Step ĝs§ĠÜ"+Å
hÆ

 2

/Ĵ 2Ø@AH[$

¦ó"Ć

æÁ

/ĵ2b)p$<N[eäï0/S_AZK-Íĉ!ĭ|İ RNAĝs§ÑeÜĉ/

¼ÛĂĈ"Ě/@AH[#ĦñĂĈħ=Z]M;f@^dáÞ3¬/

"ÅnzĂĈÜüÝó"$Ó'#¿ï3Đm«

/



Æ

$ďòĆ

@AH[3ïĂĈáĉÓ'#´ĦĴhėĵ

Ĳ§Åĉ"ô©ó¿ïõü+Đz`WDf[#~0/0-#õü{"pi"ăo









!"









ĠĜÉó"4ZNĢ3ÿ¯"ģ/`WDf[$îx"¨/ÿµ!Ĳ§Å
Y@fd.¡ß!Ġe¡ú#áĉ§3*ĭ£Ĳ§Å@AH[/ 5)Øû$ÔĮ#ğhÂ

0

Èå#öý"ÌÇßĖ¸Æ/ 6)

!¿ï"`WDf[ģĉ!#¡ßÁ¢!è"!

N4@aå

LdBdA>SbG@^då /!ĭ£`WDf[3oģĉ$£`WDf[$}Ĥ&ãÛ"Ï#¿ï

+ĂĈ#¿ï3Ë"Ĩ$¯$4 =Kdå#ï",.í("ß/öđ0Ė¸#/¡ßÁ"±$ĭ¶"°®/÷0

/{ċĎ`WDf["

¤(

$!

ć

#ê½ó!ÞĘ

Ēĕ0/éÁ#¡ßÁ-ĭ¶"ČçÖ(0/©ó4Z/#"±lĤ%v"4ZN3Õ/4ZNĢ$¢-0/ 7)lĤĥ#ð!/$é.#²å#Ħñ/

VSFKø¿$`WDf[ A >5J"Ą tRNA #ãÛÁ4ZNP >5J"¨/t»VSFK×4ZNĢ#8aWMa3ãÛÐÎ/
Df[",/yï$l"Õ!Ġ",/7dJcQfÚĕ/",.ºĕ0/`W-0/ěøëÄª«"À20/7dEaQfÚ+zÊgÒtRNA "Ą4ZNĢ4ZN#

pKa $ 8.0~8.5 ù¹
áÞ¾

ć

ć

.kÁÙr"-0/Æ$ýxó!Ĭ­$!4ZN#vą"j4ZNĢ#²3ĩ­/kÁÙri"/ìó!ãÛ¿$ãÛÁ4ZN#ĊScJd
//ĊScJd#q«!2ãÛÁ$4ZN,.wj

ScJd0!

PKc9@x$x-ĸ-+`WDf[#"/0#$!



kî

0

¶kî$çèå

Ü"

04aMc?

S_G@[aĈ3Ê»o"

0´Ñ#ä«y¹3¨Ă0*pÿ3² !

{Ý ! OJ `9 @S `P 8a ú(HPA) 3  Ê4\Ka(Ala)."#o0;]@a(Gly)
      

OJ`9@øú(HAA)3=aI`c^Ê (Ď)3ùÀ tRNAPheCUAµ"zÐ"4WL4@^zçè DHFR111 Ë"Ì§r UAG 4aMc=Ja&#r3ê·lÎÈ Wild Type DHFR 3=aI`c^

<^t»üâ.ryÇ3ÙvGly |%OJ`9@ćão-HAA $yÇÐ!rÓí1fÿĀ#ĀAla $ non-acylated =aI`c^
!d®¶ÒØÊHPA $~20%#~ÇéÏ&#rÓí1OJ`9@ćão$o+]UDcX#
0 mRNA HaS_cI3ÊÖ(<5%)#MaJ#)Ä1#°Ó!r$Óí!01."8]>TSFJ3=cJ

4aMc?S_G@[aĈ3ê

r#Ó3ê67BEaR`GH5a;#à¹

MALDITOFMASS "-0½Ala $#rÓí1!

HPA $ 20%Ö#~Ç#rÓí1(MASS Ä#à¹.r1
Ą4WLú$HPA 

0



1

je#à¹-/TSFJÕ}¿¼.
/0Ô1!

OJ`9@ú]UDcXû#"+zÐ4WL4@^zÃ3Ê
º#no"$-/Â

OJ`9@ćão4WLú-/+r

1,

ąÏ

à¹

0ryÇ3ĉ*0210ÍÁpÿ3r0"-/.!0ryÇ#e¯b½#x(10¶à¹$
Ą#ÉÐ
³
Ć
Å

"

0r°."!
!



q"ò'





/#ÒØ"Ð£¦.10Æ"­
0d®ă!#éÏå
0*"$ÉÐ$éÏå0

/0æ

¸
+"

¸"

0



0!HcV#Č]UDcX¢¤ý3Ô.#4S`cF3f¾"©

0ÉÐ

¿æ#g

Ü3¬½!Ą3û0@BHX"õz­!ĈÚtõzÜĊC_:@[aÜċ#āÎ+(Ć0@BHX#ĈÚtõz(in vitro selection)3æ"0¡ë#¼Û"+¥0soÐ"$±ñ Ribosome Display Ã 9) (öm

DcX3iìÈ
3+

3ð

¤ý"!0

0TSFJÕz}¿¼

 Chemical Probe


0­

0(ó"h#Þ!ôïÐ(phenotype)

"āÎ Read-through Ribosome Display (Rt-RD)Ã 10)

aNcIYaIzÃ 11) (IVC Ã)3ß)2
tRNA # DNA ÷w3þ

(genotype)

Ã

Griffiths ."-āÎ1=0

DNA # 3’µ"QZcA[a0.č×ć# RNAĊmRNA-tRNA

0 mRNA e"]U

0EaN:3ªÔ0 genotype-phenotype coupling Ã)-% tRNAċu10-""-d#þ

DNA0Ċ 4aċ(ā

=JaĊATGċ#®" FLAG-tag 3=cJ0÷w1"á4aMc(TAG)=Ja3-ď-¾]Ö#wÂÄ
DHFR " Î^ y 2
6RB"Ôr DNA 2dQ¿Ø!
/ tRNAdcq

. ARS !ÆÈ/(i)3PER7GO?7R"¥
2±$(ii)3P
ER6BP#MRB
9 N R  / (iii) 
FLAG-tagMH;

RJAPCN2
 mRNA S!
±/.(iv)
  Rt-RD/IVC  tRNA  in vitro selection

[¼'.ßp 4bà3PER7GO?7R¥". tRNA !Ã"Ôr DNA,d/ mRNA "'2MRB9NR.)IVC ¤2¬." Rt-RD/IVC ¤2*!
tRNASerU "3P>6BPNRGÏ!LP=JÑfn(GRN,:O58KP2Å0Þ! CUA 3P>6BPU74B!xrÑf2n( tRNAtRNA !#v¡

us`Ü/."+~40%´& Enrich /`Ü*xrÑf

Í/-in vitro!

.h#tRNA "ÊiÌ´2ª.S*Û®½.
7GO?7RtRNA ":O58KP# in vivo Å1/¤#£§

 Rt-RD/IVC ¤#in vitro!

.:O58KP2kÁ!

*".°·±/x ARS !ÆÈ/.7GO?7RtRNA "\7GO?8KP!
ÕT.µ"ÓQ<PF5!~*in vitro"h©q¤2¬3ID38N:O58KP¤!

.1/.

³ÒºßRSà" in vitro Ý¸cÊi!.MH;RJ2W¿89@J#Ú !¹a/89@J.©q&!Å1/¡ °·2**¯"Ïe v¶»Z°·*&&v"¢2.©q*v"°· ÊÅW-Y/,"2toÇÉ °·#XÐwzwz×z°·²lQ«¨izÙ°·}Å1/°·-.Ù{|!¦­S&&°·ËÅ!jgbm°·ÀV%!z«"!+-¦Ç&1) _ $ Aoyama Y. Chem. Eur. J. 2004, 10, 588. 2) (a) Sando, S.; Narita, A.; Abe, K.; Aoyama, Y. J. Am. Chem. Soc.
2005, 127, 5300. (b) Sando, S.; Narita, A.; Ogawa, K.; Aoyama, Y. submitted. 3) Collins JJ. et al. Nat. Biotechnol 2004, 22,841. 4) Sando, S.; Abe, K.; Sato, N.; Kanatani, K.; Aoyama, Y. submitted. 5) Budisa, N. Angew. Chem. Int. Ed. 2004, 43,6426, and references therein. 6) Sando, S.; Kanatani, K.; Sato, N.; Matsumoto, H.; Hohsaka, T.; Aoyama, Y. J. Am. Chem.
Soc. 2005, 127, 7998. 7) Tan, Z.; Forster, A. C.; Blacklow, S. C.; Cornish, V. W. J. Am. Chem. Soc. 2004, 126, 12752. 8)
Ogawa, A.; Sando, S.; Aoyama, Y. submitted. 9) Lipovsek, D.; Plückthun, A. J. Immunol. Methods 2004, 290, 51. 10)
Ogawa, A.; Sando, S.; Aoyama, Y. ChemBioChem in press. 11) Griffiths, A. D.; Tawfik, D. S. Curr. Opin. Biotechnol.
2000, 11, 338.

-á-/13987,:#2 ;*  !-   "6
0$&5'4)( +.%        >,E; âę±>¶:±Āā;¬5&ú»ðõ+ßå:ÕĆNÅ31º®ăÕĆNJ-
ß;ÒČ>Z~X|[Nĉ;-K)8;HJĒë=1E=m{Wwr8ÀILKăÕĆ=Á%¢ù7J)LD7Ö:±=ĜÙċGfhÏÉkPlxX~Zv|=1E=ĜÙb|mz~d8+6ã*L6&1ñô%Kâ§óÅ=1E¶:±ĀāĈé80=òÑÀIL
KăÕĆ%¹ÏÉ=×ù8fhs~dyĕ«7ü¤-K)8;HJÊ1:×ùNÅ4fhqbx
Qy÷=%¢ù8:36

K ¨Ĝğ1,3WyT|=ÕĆĚZ]kRw|>Ď=ĞWyY~[;Ĝ4=¤7ç;ĎWyY~[NÅ4T~ew|>ĎNÅ1: ě8¢ąç:ĞčI/O¾ÃĈé @ĞčI/O=Ì³´ÕĆ 1,3WyT|Ė>WyY~[%ĜĞò¤7ćò+1ug~V:ĎÕĆNÅ2Ë:ÕĆNÅ31­îĖ7K)LI1,3WyT|>ĎĘÓ;äÐ-K½

I/O¾ÃùNÅ2¯

7>ªÈ=8L1£¼&ĞčI/OÕĆ8+6²©-KD1ja`G¿:9=ÕĆiws~^>ÒČ=0L8BC¥,7JD*;­îNÕÃ8+1¯=ă±ÕĆ7K8ÿ"K¯ĞčI/OÕĆ>  :9=ÔÂÍ×Ý°7

1,3WyT|Ė=ÝÛßÂ>*I;û¦Ü>ē   +ĞÎ=w|_rYSyĎ8:K%Ý°NÚ;öÇ-K8@=ĞčI/OÕĆ
Nú»ðõç;Õí   -KĚ¨ĜěÌĄØI>1,3WyT|=    Ĉé;Kê=ÒČ%²-K8­îĝÎ8ÒČĜÎ$I:Knb{ĞčI/OÕĆ%¾Ã*LK8¦ÜáĂN+6K )=òÑ>­î}1,3WyT|%ÒČĖ!û=ÕĆÂ8ú»ðõÂN<"1ug~V:­îĖ7K)8NĀā*/KF=7K1,3WyT|=ú»ðõç:I/O¾ÃĈéNU~m{\[8+6ã-L?­îN¬8+1'Ê+fhÏÉ÷=þ%¢ù8:K>.7K
­î}ÒČ¤$I­î}¤Ãę±¤A=·ď 1,3WyT|%ĞčI/ON¾Ã-K:ė>îĎĐ=æÚÂã8ø"IL6KµđÚ;#(KĞčI/O=Ì³´ÕĆ7>I/OĊ;æÚç:ĜÙesS|%²©-K)8%ËI$8:36

KæÚÂĜÙesS|%ÚÂýĔ7ML1°ÕĆ>ZV{cU[dx|=TwràĒë8+6Æ"K)8%ÐK))7ZV{cU[dx|>   :9kw~
z|Ė=L1o[d8+6×ù-K)8%H'èIL6[dx|=ĜÙċÖ=×ùNÅ36

KF+1,3WyT|%ZV{cU

L?¥,ÞïfhÏÉ7KT~p|fh`t~l%ĜÙæÚìĐ;¡JDLK>.7K1,3WyT|}¸T~p|fh`t~l¤=¾Ã   1,3WyT|Ď%ĞÎI/O;¼&ÄKĈé;ÒČN²*/K81,3WyT|}ÒČ¤%--­·?>/z­·ÕB$/55
 t8ÿ.;=ÝBÁ
Âă¼ê

%

 .ü´

)6>+.

% /Ô½Ö.1,3LiJk/   ÙÖB$/55¥Ø*đ%û³ć.;=ú/1,3LiJkBĄÄ.Ù¡BÔ2+ê»z/  .;>¦Bñ'%$/éÊî×+.1,3LiJkÜ.óòĊ*/â-]Tmk­·Bì

)>+ô>z/čÀ/aj_DGi;=­· òĊB<#C

PS_FgkÛÅ/ÆÏùY[Uem`òĊ*9Þ?)>+Ã<+-')>Ďđď1,3LiJk/¬<#C­·ì
.

²<?%

%z/±0 

*=

¾Ç/\kXi/òĊ
B1,3LiJk<#CÜ.ó)>+ Ã<

+-'%/;.

1,3LiJk0Ìÿx/č¢+9u{ßđĔ(a) PS_FgklSWNT z/ÔÙÖ+gV^kLÐª/Ñª1,3LiJk.àÉ!>µB¸'%z ­· (b) z/ AFM !>+å?%*1,3LiJkĒĀ<#C/±ç  *>+Bë¶!>+1,3LiJk0$/sāÏùBËø.#>+.;=¾~/±B¸(  \kXi.©w)>+.->
1,3LiJkl

)?

z­·01,3LiJk/fZmK-gV^kLÛµBÃ<.

>(5=
ĎĐďz­·/s-Č0áÔµu{ß*>%8z­·0MQWchdm/µ.3+C,¯ċ?-Ďđď1,3LiJk0MQWchdm/OGR:­Ü.A#)sāÏùB#>%8”  LiJk”/=6ì+->/;-Ûµ01,3 x/áÔµY[È¿.§$/ýæ£Bº| ĐÓbQW+)¹>ï

ìµBå!>9/*>/{ÎyöB¤!>%8.1,3LiJk/ĐÓbQW+)/ÒìBÐ-Ċ<}%xp$/(/·ÊBèv!>

1,3LiJklchEZhkz/­·   5"

 Ð*áÔµ/¨Ĉµč¢*>chEZhkBß)1,3LiJk/ĐÓbQW+)/ÒìBÍ)6>+.

%ÔÙÖr*0ĆlqÙ!>chEZhk9

1,3LiJk/ĐÓæ£.=7+É?1ÔÙ*>+Ä.æ£*ĐÓþ
*>0"*>¤ąPS_FgklchEZhkz0ÔÙµ*=ÙÖ0chEZhkÛ
Å/ÚĉðB%5%ÔÙÖB  .;=¦%+@Ă 

± 

/_DG\mÏùB¾äõ!>+*%ĎĒďNkWgQW0chEZhk.àÉ!>+ë<?$/Ă0ß%PS_Fgk+34ní%?<0chEZhkPS_Fgkæ£.=5?  Ó.þ
+Bå

)>

)>/éÊ0o/{ÎyöBÃ!>9/*=1,3LiJk

ĐÓbQW+ĒĔ(a) PS_FgklchEZhkz/Ñª(b) z/ TEM )Òì!>+B¬å

)>



1,3LiJklchUI_Hkz/­·  :chEZhk-,/-chdm&*-Ëø-sāÏùB¸(¨Ĉµč¢.()
9B6%chUI_Hk0Ëø-sāB¸(¨Ĉµč¢*=Å°Ă/.;=®-ð®B«÷!+ã<?)>ē .å!;.chUI_HkPS_Fg--g,ĊÉ|Ýÿ*709"ãÃÐÏ,êćê6CfgK/(8(¼6()$"1,3FeDg(,*57\dQCYAgcgP^H?e6ýÒ,HgYB_iJ
ag/("(;Ú&80"

ĊÉ|ÝlºÓ'8(;¾&ªðò 3£9"ĈČ ĉ1,3FeDg7lºÓ;¾&\dQCYAgoýøÝ
'+9&8(å6986*\dQCYAgĊÁ1,3FeDgÝ'Þ¸98"2Ên'3Ć,āµ9×},Î}-£9)Æ)ïñæ;·%ząJN
Y>cg;ĊÉ|[LT(&Õ8('Ôv

;OiGRT(
"MgIiá4
(&,°ÕÙÜ

Čď JNY>cgh\dQCYAgv,«² v,  LZETeÀ­98zą1,3FeDg;ĊÉ|[LT("¤ă±Ć,óØÈæ     4\d=Wdg1,3FeDg,ĊÉ|Ý*709"ċuÊú{,Ą'GLT\d]i(qwÕ8(å698j»ĄĈJNY>cg'-{ýFeHiL

1,3FeDg-oýÊ±,1,3FeDgì

DiUcg'-ĎuÊúĉ-GLT\d]i(,qwÕ*-Ām)(*)8ĈċÇ¨ĉ1,3FeDghÄúv3Æ,vÆ¨;($&8(å69

*Ām)JNY>cg{ý/,æ¤~÷t¤~,þ×*¿'8(¡9&8Ĉč ĉ3 ,5)zą1,3FeDg1,3Fe 
DgÆ ÖÊ±Ć,ĊÉ|[LT(&Èæ9. ¯698v/,óØ)Èæs
 !' JNY>cg,FeHiL{ý*]gX 
iL;zą"JNY>cg;Õ   8- 

OH

(a)

O
HO

HO

O

O
OH
HO OH OH CH2
HO
O
O
O
O
O
OH
OH
OH
OH
OH



OH

NaIO4

O
OH
HO OH OH CH2
HO
O O
O O
O
OH OH OH OH OH

RO
OH 1-n

HO
O
OH

O
O
OH CH2
HO
O O
O O
OH
OH
OH
OH

O
OH n

OH

HN

mæ()8-'8Ĉč ĉ\d=Wdg(,v«²;1"!,ãÃ

O

O

O

H2N R

(b)

NaBH 4HOO
OOHHO OH OH CH2
HO
O O
O O
O
OH
OH
OH
OH
OH

O
O
HO
OH CH2O OO OOH OH OH OH

R NH
HO
O
OH

O
OH

1-n

O
OH

n

&!

"%

7<#GLT\d]i*fEQgïñæ;s $m'8p¼6()$&8,5* zą1,3FeDg1,3FeDgÆ Ċ &'#"

É|[LT(&Èæ8ãÃ¯69&8r čď õb@áúú; Õ"JNY>cg/® Æ)Èæ±VX]Sd=e;í8k' ,ö¶ØząË  zą1,3FeDg;
Õ"GLT\d]i/,óØÈæsm,5)óØ)ÈæsmÅ2&¿'8(-î0'3)
1,3FeDg;ĊÉ|[LT("ÑÈVXà,ĊÉ|âä   ûURT4ü¥VXà,ĊÉ|ùv-_`di4¹Íá(&è·"9&8ô§96ÑÈVXà;ßy*ĊÉ|âä8´Ë,þ×-;:Ì×*ë:9&8
³-1,3FeDg,ĊÉ|[LT(&,Èæ;ÑÈVXà,âä*
!,ãÃ


x.

Õ8(;å"¬čVX_iTe,üVXàÊÐÏ*JNY>cg,  ÐÏ;8( õÛ'üVXàÖÊÝÿ*709

ĊÉ|=fiw²'8(;¼6(&8ĈĎĉ1,3FeDgÁÂ¿8ç¦âäØ)6<«²æ;©('(98VXà,ĊÉ|âä¢½*ë8(¼6()$&8

--

jé*Ă

ČÍwZNT:ČÍw³3
 Í.ČÍwäĄ@UV³,!*~Ý#<,@5'1,3I`Fcª·#<ČÍwäĄ.k6³µ}@ktéì!ČÍwä
z+Ā³@ñ,|Æ=1­±='ċ}@°<,+<,í:=<4$1,3I`Fcäz+/ktéì.ÿ!'MBOQacô¤p@·!pª·@ñ)'¯ y×£.9;Ā@ă!'&/êÇÐÖµ/[_MBOQacÜ·#<čď1,3I`Fc@ČÍwZNT,!'āUVæ/éì

,Á:,-)*<4'Ê.B`KGLL^c@;5ČÍwäĄ+ë³@ýñ%<,­f-L_FUVXACWe°:=<,7Á:,-)*<=:/êÇ01,3I`Fc-<ČÍwZNT,!*(+-]V\e/³µ8³Ù@±+<ČÍwĀ,!*Ëî#<îµ@á#<7/+<Ďď1,3I`Fc@  ÍwĀ,!*~Ý!'Ā³/È´   ČÍwĀ+Ü·!'L_FUVXACWe/  v

?;.
 og/9.Éüµ,ï¨éìµ@¸)'çd1,3I`FcJNT}/ªØ8


µ.q

#<,-Ê-ËîµUVÅ¿@;5ČÍwUVKc[MRT@h<,@Á:,!*'¡rĊ1,3I`Fc@ČÍwZNT,#=1Kc[MRT/:-<éì8Ćâ7î,-<0$+<4'Üpÿµ/1,3I`FcÊ-UVÅ¿,Üpéì,/CcPeXDCN,!*u1Îµ8{Ćµ,)'ĉ:Üpz+/ËîßÚć+)'UVÅ¿@WCEUV\S_B`.¼#<,+<+>m¯ÄèUVSH
VbMe(+-WCESHVbMe.*7Ā-«@Ç'#,Ã®
 ÷ûďÄàã0ÀÔ¬Ò½ºnò@

6<  

=< YbMDHT/fÛ+;»¹¤Ĉ'ÀÔxÜ0"6ùĀ-ó@)'i§©å¡ ÌlÂxÜgÑÓfjxÜą©å¡ Ă¥sxÜ.²9;Þ!g4#4'àã/þñ.!*'('i§¡/āÒxÜ Ăø¦úÁ  -:2.ÀÔàã¢ I`eY/¡ÜöÏ.Õ¶÷ð!4#íõ¾
[1] K. Sakurai and S. Shinkai, J. Am. Chem. Soc., 122, 4520 (2000).
[2] M. Numata, M. Asai, K. Kaneko, T. Hasegawa, N. Fujita, Y. Kitada, K. Sakurai, and S. Shinkai, Chem. Lett., 33, 232 (2004).
[3] M. Numata, M. Asai, K. Kaneko, A.-H. Bae, T. Hasegawa, K. Sakurai, and S. Shinkai, J. Am. Chem. Soc., 127, 5875 (2005).
[4] M. Numata, T. Hasegawa, T. Fujisawa, K. Sakurai, and S. Shinkai, Org. Lett., 6, 4447 (2004).
[5] C. Li, M. Numata, A.-H. Bae, K. Sakurai, and S. Shinkai, J. Am. Chem. Soc., 127, 4548 (2005).
[6] T. Hasegawa, M. Numata, T. Fujisawa, K. Sakurai, and S, Shinkai, Chem. Commun., 2004, 2150.
[7] A.-H. Bae, M. Numata, T. Hasegawa, C. Li, K. Kaneko, K. Sakurai, and S. Shinkai, Angew. Chem., Int. Ed., 44, 2030 (2005).
[8] T. Hasegawa, S. Haraguchi, M. Numata, T. Fujisawa, C. Li, K. Kaneko, K. Sakurai, and S. Shinkai, Chem. Lett., 34, 40 (2005).
[9] M. Numata, C. Li, A.-H. Bae, K. Kaneko, K. Sakurai, and S. Shinkai, Chem. Commun., 2005, 4655.

--,. 0: 53( ;#+  & 2 !4  % /67 *, .-9 -$   '8       






 óĈ3%Bóè5ÕČ4?2ÁČ/52$À¬ızô3?,.îõ

û3ċ:ă.@CBĝ¬ÞĢ4ÂÎ3?A°ð(.Bğ¾{ºĈ3.<öäÊzôäĉČĨČĸķzô21ß2¬ızôEĽ(.·'2¬EĴā'*B:"¨$2'C.Bî3òï¬0Xqgo¢¬0EâÛû3ċ:D*+ÞĢE<-ípWLPq =®û35Č(.2ł~u4òïí"z3Įï3ģČ(.BíK[^q   21íñû3č:,+¬3¨©4ĲÆ"³*@C.
B&C@4ĝ¬5¬ĉ¬0(.ĒéÞĢEÕ(.

AÒëÈąØÑ0(.ÖÃ/#Bĝ¬3ľ¿4âđE}w)B35ď´2¬Eĕĝ¬EÞÎ)Bl_Z]3¨$4Ë¤EHqfZ])B&0"Ī32B{ºĈ3.&4?2Hq[nRIq]¬EÞĆ)B&05¨$4ĕĭęEÇ0)B"óĈ5ńŅŃ=ĔúĄ4óľ¬Eô.Ķì$ÿ4âđEùð'*.B&C@óľ¬Et-4l_Z]0(.{ºû3ċĐĴā)B&0"/#C6×Ù4âđ3Ò+2âđE}w)B&0"đ32B0yÌ'CB ×ýĂ/5{ºĈhS]¬?A<?A©#$ĵ2¬Eîõû3ĘĚ)BÐEĝ¬ÞĢÂÎ4+;4ØÑ0(.ç(+
9+Ð"Č)BÐ¬Eºª)B&03?AÐ4ČħEÒ+2âđÊÈ¥0(.èô(+×Ā/5~v4-4]dZ

MS3-.Ċ|)B  K[^q4¬ıĻÅįHjsRr Ð4Ďïĝ¬ÂÎEô(+aHJUqQqNr ÐEco\GqNepZM0)Báïĝ¬r Ð3ĦþĬ¹ĭE¶(+Ò«

#

1)4

"

+ ð¡óĲģí4ÜEû3ĩĉÐ 21"aHJUqQqNĉ¬0(.ô'
C.B"¨$ĩĉUqPs5)/3ê=ļºÝ21/°ô'C.B(!(ğ¾ß2ÎíEÜµě0)BÇ"AÔ­4ĩĉ/5ĥÈ/#2tÓÐ5
A0@>BWsOZ]¬Eîõû3Ü)B&0Eđ3)BØÑ/BÐEô+÷UqPs4x/<ĹàĚÐEô+ėĺfmTkqĿŀŁ÷UqPs5ć!-nF
oWHi/4Ü"đ2ĞġÓæ/Ağ¾Éġ3Ĥ¸(+ÏĖ/B(!(  EÜñ0)BaHJUqPs3
3

.Åī4¬EÜ(?0(.<¬"UqPsYZfu¯'C+Ð0Č(+0#4QN^oÁ¿5Ĺ¼3·'&C5aHJUqPs4Èą"UqPsYZfėĺ/4´¿§-9Aĺā+A4Īī§3ã)B+;/¬EÕÍ»0(.Ü)B35QN^oE¦½'*BÇ"ÕB×ýĂ/5Ð0 

ÊÐ04



ĝ¬ÂÎEô(+¬ÜQN^o¦½æEİù(+  &&/5jYocJpOq4ÜæE3Ġ8BjYocJpOq5®3%Bķ¬²

0(.?$ô@C.B9+ĳē0(.<ô@C.+ĺbsLqVqøE7#Ĝ&)í4t-0(.<ü@CBÕ±í/BÐå¯£  E÷(.jYocJpOq3µ(.îõû3Č)Bk`Mps^oÐEÄ+(!(&4ÐE--¯(+UqPsYZf£Ú)TGWM<X@Y8¶r%1Ï*TGWM<

+

+

H3C

N

N

+

+

H3C

N

N

CH3

X@Y8­o5'+}Õ&"!6+k)Ë
-!2)TGWM<X@Y*1**pÏ

1(CH2)5COOH

!0)L;<DYAZ*ÀB?JW')È|5TGWM<X@Y8¸»½)

H3C

+

N

N

+

+
(CH2 )12 N

+

2

N CH3

#Û&­o5!0)+fTGWM<X@Y)Ãy!'8(B?JW'%x4o5*{Ø8l¦5B?JW´*[#f'  hv'*Ép&5f8~  TGWM<X@Y)5f8gÅ5!0)º!v³  '  hv&
5M<X@YÇf !DYAZGHO)  hv  'f8×±
a`)¶r5'  *ÀB?JW+v fÓ*cyf*Ò''1)5&  hv&5M<X@YF;QZ *b74)TGWM<X@Y8¶r5'DYAZGHO])6!Ép*§)5f*ÃyÎd)TGWM<X@YÃyM<X@Y
F;QZ'f*ÉpÔ65ÜC>ZS Ý TGWM<X@Y*pÏ 
B?JW+pÏ  \*f*ÃyÔ
%Ï*TGWM<X@Y**Às)Ä¡75')(5#.4*BCIS)Ï+  hv'f'*Ép8Ô!ÏÜn(

Ép)$B?JW3*·ÏÝ'%UKEZ65¨BCIS+ep8
f©)¶r5uÂÁ)²Ê%ß¬À8&5'7"! 






+




(i)



(ii)
+

(a)






(b)
+



+
+




(c)

C>ZS f'  hv8º!epÛ­oBCISTGWM<X@Y)25f M<X@YF;QZÐfÔ'f M<X@YF;QZÉp fM<X@YF;QZÐ
f8!DYAZGHO)M<X@Yf'M<X@Yz¥AYOW8¶r!'*¹i'3) *)f M<X@YF;QZÐf8¶r!Áii TGWM<X@YÖ ^&*f M<X@YF;QZÉpÐf 2,TGWM<X@Y ^ '  TGWM<
X@Y  fÃyÎd   TGWM<X@Y  fÃyÎd         C> S  )¿2)  f8«)tiÚ!  f8p'%vfw)2"%f8ÍÃ!£°¹Ép8y!º!f+=G<YPWN:VYÜ ~

Ý8m¼%36!f  '9K<YPWN:VYÜÝ)Ãy5f  &5f )+9RY=HOVY?)24PWN:VY  8tiÚ! 'tiÚ  8µy%3
6!Ép  *¯Ì8vÓqÙÑ)24!Ãª **Þj*¢¹¯Ìf*¾Æ6!

*1**¸»+e '  *_PWN:VY)Ãy5[¤  + )Ãy )¸»½)Ãy563  8¯p'5°¹fÉp  +  )+nÃy )*/Ãy!#.436!°¹fÉp*¸»+®0%Û

--

CH3

'4#E"0. 

N+




IgM

NH N

+

N

H3C

COOH

N HN

3

N+
CH3
COOH

N HN

HOOC

COOH

NH N

4

IgM

COOH

 G1CH3N   RMf^ °º¢pÓu  7¡µ  ¢]c\Hbd¢p  7¢z´3CLTKd]

c \ H b d            G Y K d  ] c \ H b d     4æÉ£Ñp     

H3C

+

N HN

+

N

+

N

NH N

N

CH3

5

+

CH3

 '75°º¢páÃ¯Ô¸7ª²Îã68im(46¾5Cu6¹¾À6È+C¢z
È×oFØÉ+C'4#Ë3C¥7È×oFÓuÎã6Ø),C'46@BJIcRÅ7q¢z4%È*å

3¢z7

F®Á+C'4#3$C@

65C1;BÆ§eu­Ì4*27v½#«3$C¢pßF15&¢zu6ÍruF½C4å
ËÍrI_fQdO¬¦65C;.°º¯ÔF¯¡+C¢pu7àß6Ì¸Ftx),D9Ì¸¤?XaUOWbZbfPRV^Å7±Ë¬¦4*27v½>«3$C4Ê!2C







 l;36 4 7@

0%'8- &
0%4*)66$+

 ¡ ] c \ Hb d 4 ($+,  C  8 â ~  p&F  © 6 Ð Ò3 $ C ¢&2/$!1

p F  C ' 46 @ 0 2
¨ . 5   C  8 r



-.3&

À4*2¢p ]0%'8!9


c\HbdÞp77":
*# ]c\Hbd4C8â~pFs6}B=¢p6«)DC±Ëeâ~p6+C¹¾[11]ÚyË[12]F®Ï*2$.¹6¼pt6
p4Ü]c\Hbd47



0%5-

± Ë F ¿ » ), C ' 4F+*5]c\HbdçLTKd]c\Hbd6È+C¢

p8ÚyÙÇ7g1 Horseradish Peroxidase (HRP)4än7¶F>/)A6å·7ÕÚy³Çky¸4*2¼pt68
p8ÖÂÛÞp7<38h3>-7¶FE5'4#E"0.¢p#È+C*5ÖÂÛÞpF½D9-7ÖÂÛÞpSd[NAD5@5`YfN5|45C4Ê!.ÖÂÛÞp4

7ujuF}B=Ý45CuF+C'46@Bå5|w{:Äp¹¾--Ìz}Ú)(7'Ø587«ÍÑ&+=BXZ([G>Vêh õ ö)7WQ
C[]OZh;iß58"[G>Vêhh~h$´Ú;°à" ^æ*ÕÙ¢i)46[G>Vêh;ÂÊÌ)×~7  Ðò*WQC[]OZh58"8O
5*h+n%  &#"=ZE\î¶`Y\éP +HO
RKS<]bh  '[G>Vêh  ;¼~
ClO 4 RhNP)=XP\róh&7 =IM=UN=CYZé;O
'%½t"*¿»)·Ô;RTY\D&  ©ëz "zÆÁ;l¹AX
6
V;Ç"   4-  )%p®"hð  x¸ '%Ç"=BXZ([`&+ =IJZ=XP\ h÷h*XIUh G>Vêh 
587*)%h`&+ h*0Æ%7':#"õ öµCOOH
ä'%   7+d*  h'[G
H
H
2
H
COOH
>Vêh  '*¼~¿»;Ç%³*­e
H
P
Rh I
NH
H3 C
`)% =IM=UN=CYZé*·ÔPO
NHCOCH3CH3uz;Þ#"'985*zÆÁN- - L -+.% h' h*XIUh&#"
 
> 99 % ee
85*×¯5[G>Vêh  'h  '*êhÓ)7?O\J@ç+[G>Vêh«h*×~èf){6/87  h  )×~"[G>Vêh  *?O\J@')46+2%ËÃ8"3*&7' çÌa¥·Ôuz
:#"/"h×~7')46[G>Vêh*Ú+_"«h  +[G>Vêh'·¿»b)%(êh; ÂÊ;ª7±2%y(a¥·Ôu'(7'¨5)(#" «ÍÑ)%ÆhÓ*k8"´Ú;cÓ)Ö01')46§´ÚËÃFHLV*²Ò;0"ÛÏ2"ÀÚs;cÓ)/{6m8%!*q;vp)o8,«cgpÆhôp~¬¦+Å*ãÝ;¾&7Åá¬¦'%¹Ç&73*'Ø%7 âå «ÍÑ*nÜ):"6ñ%6/ìíÄÍÑÎôpÎ£xÈ¨¤¡)*;j6wÉ_/[1] Harada, A. Acc. Chem. Res. 2001, 34, 456-464.
[2] Miyauchi, M.; Takashima, Y.; Yamaguchi, H.; Harada, A. J. Am. Chem. Soc. 2005, 127, 2984-2989.
[3] Oshikiri, T.; Takashima, Y.; Yamaguchi, H.; Harada,A. J. Am. Chem. Soc. 2005, 127, 12186-12187.
[4] Yamaguchi, H.; Harada, A. “Macromolecular Nanostructured Materials” Ed. by Ueyama, N. and Harada, A.
Springer Series in Materials Science 2004, 78, 258-272.
[5] |ºïxÈ¨ 2005, 54, 825.
[6] Yamaguchi, H.; Harada, A. Chem. Lett. 2002, 382-383.
[7] Yamaguchi, H.; Harada A. Biomacromolecules 2002, 3, 1163-1169.
[8] |ºïxÈ¨ 2004, 55, 249-255.
[9] Harada, A.; Yamaguchi, H.; Tsubouchi, K.; Horita, E. Chem. Lett. 2003, 32, 18-19.
[10] Yamaguchi, H.; Harada, A. Top. Curr. Chem. 2003, 228, 237-258.
[11] Yamaguchi, H.; Kamachi, M.; Harada, A. Angew. Chem. Int. Ed. 2000, 39, 3829-3831.
[12] Yamaguchi, H.; Tsubouchi, K.; Kawaguchi, K.; Horita, E.; Harada, A. Chem. Eur. J. 2004, 10, 6179-6186.

--31

.('/62 5

148"$-70*,+!)#%
é ÚoËôđćQ^YRNZ)Ä ¿Ě ¿¿?d¿>h79 ¤æ÷ċÛĆO\^VS:Đs.IĚ n?%6>íKG
C/5k¥@¦$ė?¹>@ĉK
2Ě«Á?Ï>­CIC/5)Ě®âb?uó?!ė";č=GĚ~ó?áÉ>@Ð?äq>ö.39/
C$C/9Ü/#GC3M:/54?a¾ĚQ^YRNZ:@ Lā&H»?~ó>EGĚÚoËôđć?ãå°ÈÞù'EAÇ?G¾>
8$9Ö =úû)øKIĚÃ¨C:ÑÞ=þû)ï$5-;>¯ý1HÌé:1k¥:éĜ;=HÚoËô
đćċnüÒDĚègØ@ëěr:/5)và>@(=G?Ó±:Ěk¨Ěõ³ãåó?ßçď;/9·,Â&?Ô$Ê;=79ø+;¬KIC1ÆQ^YRNZ@ ¥lÇĚk¥:Lă&C1)Ę©Ç?îð>8$
9@üÒÀĒLÕęĚzÅ>@ìj£ê>EHÚo[U]ãåĘ  ę)e:#75?>/9Ě¥ ¨ > @ T^ V P  Ě . F > Ã Ą @ĚC:Ā;/9ĚÚoy?j£àº_Ëô|ªLµ/5ãå)<
M<M&9*9$C1Ę   ęÆQ^YRNZ@p¥Ě~ó»>1HÞùm»?.)æ
79'GĚk¥D mĘĕÞù mĚ
YST`Þù mę?Þù)#GC/
5C5Ěk@ ¥LË>ĚÍÿtÚĘf¢¤æ}ĎĚÙã å ² ę > × { ü Ò !   ( F   
Bċn?Ċ;¬$x"L'Ė$/Ě
~óa>@ĚÚoËôđćċn?ÎL¶GąG=)FĚÇ?$J$
J=Þ ?¾L07+G;ò&HËn;=GC/5Ç¥?é Q^YRNZ@iČ?ē¡tÚ?es:øKIC1k¨DĚ½/$ãåĔ?Đ´>ñ)HE%=¼½=ãåÞùĚ'EA»?~;ÑÞ=ÝúûLÅ§/9'GC1&
Ĉ+=GC/5)Ěé ÚoËôđćQ^YRNZ?wĒLX`ZW`Rc>Ą ¸Ă/C1Ę    ę







+

'

"  

%.*

+

)



- -,0/&#!
 [¤ê'.*$ æ +Xä(6ñ)3Ô Ö]È'2~ ÷'3

²ð

e*éÇ

.*i(Æ4 `+×  6í! G8SJR*éÇ  d1|½'¶*± dï,4#3 4.*+l$2 &*éÇà~ #.'r$"! `À£'1~ 6î!nË(+ ¯ù´.ª§30(¶ó%#ÎY#! £*!-jû6cZ(Ñ# ÿþÐÕ*¦ÞVñ¥ ĀþÐÕ*Éu¥ āþÏç*52/ Ăþ
j#3 ¢xë·+

V<PUC(3¢ ăþÜ j 4 46  Áö$
ðe(wpÿcY ~ °  ¾cZ

 ²ðe*éÇ (#ð1* j.¹-#ü ~ ¿+*_*¿(%"#ì j('3

*»e(ðeéÇ *f62*nË®(#ú!1 $3²éÇ ïß(!2vú!ÍÛÝó ¶g({¡ÎY3 )$(45

23

6

¸Ä oýµzmÐþ ãÛÝy6sÌ!¼\bÂÚ¢Ø¥EFG8HQBD³¬ µ kaý^õþâåQM?WO(0"#ñá'ù *ĄKIADÓ}¢»º1*

7ITWA ÃÍ qýÒ¨»ºþÙVJW@  ;R:U6  ¼mL>C%!­EF<UN=BC*uè  ÅøýôõÊþ©h T=9Oòh6Ì![«ÂÚy}¢t¡) $1+ --,-
--

!





#$ " %

!& ãĐĖč±ĐĖ¡¤ĎØeÈìĆüvþbð²(bALTC>TQ)

W·þ¢[čV_ęÔĀ¾ďÂ č£ĎďÓ ÐčĎĎ%&5?OSIKGQÁ1%'#ėÀđĕđĖ¼čā'ĐËđ¼(¼á$ó

.j(ÑÊKGQ»%

Ď

[$P

OF>CWð²yrë8W«'ÎÚ&öôĄ$($)&

%­.`(ALTC>TQ$)Õ×ÁÈv4ÕÁÈv.$

£nĀ(õ(iØº'úďđ¼ą(¥-yrë¸)ĖĘ~čõĖ~ĆgĐĘ~ØĔĒ~Ď%b(1%'%5ó[$. o¼)³©õÒ8ēaď°b\¼)³©õÒ8Ēa%¼á$ó

.o¼(³©õÒ$).âlÉYiØčZćĎ'm×BCGNęn4ùnĈ|e.$%n¯

E:HQ$9RCG

PF>&uµ´t8ÞÁÈn4ùn8pÙn¯m×BCG
N,(¬Ù'!

#8.é#ßÅñXiØčäÌvæĎ4) <P@<=BNÿd(w}Ûă§±'"¹
GN(Æå
Ă

%

&ùnBC

E:HQ$8ċ. ÿd8pÙ]Äăe(Ø±%(9RCGPF>&ā:<Suµì'!
[$.òx®iØč£ć×Ď4)

6¦Ý(¨Ċ%xØtÕ(Ít'

E:HQ$¦Ý(MIPNB2MIPD;PNBď

MIPNB(Ít'6¨Ċ2ÝsÃkî'Ć6íÏVïøziØč^ćĎ4)Ö÷(¶f(E:HQ$õÒ88c_  è|ESJ>8Ù.
Ø

._  è|ESJ>

(%|±ď3+748pÙØÈì(¶f'!
8n52##8c.(ªď°#¿¹(

b$1ĉ'5U5(ª{þ$(Č/b,%à5.ý.$4(º15U
#

%c#5.74)*%'yrċõ(iØ8)

0%6ÜÇðØ(º%1ç'ċwq(Õ$(8h5#½ĉ

8û-#ċ.ê¼đ¼)U





%­

6%

 nWaj¶&(¨,|k`m¨,S q¬^%=0<E:/¥e

¹§,¸$
¦º¬^

[y

~

%¤

´T

,¸$vuJW»Nj¶¼&"®Qdi`,Z^°t±R

³

BD

'!®Q

;.<F,$°t®Q,ZLp

YlAIF@-EH4

2?9FfYl·cRc{^}^





¡O%½

$x½JW»µj¶¼!²c{²

5C2F>.1G7I¾

Xhwc$²c{

\¢



)s³

X#

¹§,

´+(

ª

s³







&



$ *!x($ji©¯¦]°,¸Mz£6BI83IF,³V$MoR¤´«`m­P
`mPM_r

g,U(b

K$-(%  $#&/+ '"  .!  úĮĬ'") *,ëÝąĢĕ«NqfOAhĈÆaCslh.

Dr²ĒĘ

µ§rØæ

ÜīÇº)+8ÕäËę¹yĪ²³ā_@ĴÓĭĲÎĪĝī,²³Kqa>oqPR

qUs(Ġ:2#¦-Í-<¼'ıĲĪď¶İ

tćĮı

«ëĮ

ĳī-Í,'#$2#¤Å×Ī«ë.©õr²³ā_@D<28ô7+ĵī*.;82!=
aCslh.
įĮ¸68«ë-¯+%#6(ď¶-ëİ,69Ħďåı"'Ã-ĥ(ċ2#2)¿2Ò-Ħďå.

«ërĈÆõøĄ,69fPUsðČ

ď¶-ë-ðČ,&'ü,1:#°´Ù§«´«õøö-ßĜ÷wë,ÀĊñ<ì#ÛĔZM@q-åħ(ďå#$2#ç¬­À-ģ·,Ĩ@Tp@Nq
OX`sebW\<£ÔÛĔ)'ì#cYp[mgs4ĝ±¾ûÀ-K@n\K@n-Z
M@q,&'#$2#Þ,
Ĕ-P@XWqH

<ì#lqVhÛĔ

K@n\K@nÛĝ±@Dq¾ûÀ  ĀUq`G-Ä,Ä#

7,é¡

8ÿ=(7:9K@n\K@nªò+ĚýâÀ½-ZM@q,&'3'#$
2#Ò-ďå7ï¾û.âð('x­Ðġ<.9,ēĖ'22#Ă²³ā_@DL@BqPRqUs-ĉ{ãë, jZnğ4ðéĚý<%#ĝ±ĚýÝąðéiE^Qh-Ï)"-¾ì)åħ(ďå#$2#¢êv(-ùýL@Gn-7¨28|ěėĚý-ĎÏ¾-iE^Qh<Ï#ÒÌ-ô

"'jZnğ<ì#|,&''#$2#Þ,

ěė

chDF

NJ]sS)/:9ch<9Ěý,9ěýâÀÝÛ60£-ěL@[<¿-22,¥É9Íà,&'ĎÏ'#$2#è,

»Ą.  4ĀÑÛĔ

Ö<ì#§¥Ď¼-9(#Ěý¾-iE^Qh<«-(~y,ÂĞ<#ó=3¦%#-(.+(5'

qfOAh(.Ù-Í-¯+))(":<ÁĐ'y,Ï

ëÝąĢĕ«N
įî

,ďå:

9ë,.Õáþ-}£Üë<È ë©?o@<ì#ČĤblQjq@isOqHà,69¦ldnams.

   zìÖ-åħ(ďå#$2#}ë)/:9čă<Ġðrµđ,Ê;7:2#ďå(.

jqĩ-£Ô7čă-Úí"'čă-?bmIsNkq-

-ĮĮ-

ČĤblQ

,Ć92(u®,ĎÏ

+%&6%£¯Đ"=  ķ1ã 1[CXiçĲÄÐ1µ;

02HUlNŀ³1ĒĔý/ã/.÷ó±ý0Þø/Ň¦6,ĥēAÈø>+6
%ÜÅ1~ňīð2âv¬±Ć±ÓģĀĉMiOl1ĠĦØ÷0ŉĠ÷02w»e]JD`œĆ±ýĜ

6%-ñŁ1ęÌ-166,×ß1Ć±ė2āöĂ,=1Ŕ1Ŏ{1īðAđ+Ģ(+ņ6%Ü1īð,2 hlfT1æÊ0§*  éę1'SdiLglKci0ÿ%Āĉ)6<C_Vĸ- AELQfĐ,)/=ŋC_VCKfŌ  Aü

%A

đ+ń­

   

C_VĸAOiWIo4Č0¹"=%81 AŎÚľ,Ð,=Ō0)+ĩØ>6
aeG1Ć±ė%'
ŋ

Ċ

%

ŋC_VCKf%ÜÅ0÷1ġÛāöĂ"=CŌ:3YFld`1ŏjÝ0ÖĆ

0â¬Ā,1ÕĘÔŅ17ŋ ÔŅŌ0)+9ă
LQ`AĖ-+%&6,>%´;+%&6%č¥1ĀĉK

×ß,2.1:/ĀĉKLQ`Ü9ĝ1Ŕ$+āöĂ,=Āĉė-/=%802    ŋğĨ,1Ō   ŋĀĉúÛŌ$+   

1nÇ,=|¨0%þB2Ě/<0??-Ė%--Ë6"1:0x¢1YFld`,2\ZPTe]JD`bRfĻ}OiWI$>;Aã"=Ñë;02×ß1ĀĉKLQ`6,¾Á´A<Ò61÷2ń½0m¶0%-+&6+$%ī¼%1,ė1þB2Ě1¸ļzª,(+9ĝõà%91-Ë6"ûkÈċ9ń½0íü0Ģ@>īðĎr1s²ÚľA¬¾0İ+

66

%

ő{1~ňīð1ÅŏŎ{1^LOlüĤAĢ6
Ī9ń½0ò^LOlüĤ1ıo,Ľ®

%īð0Ãđ^LOlüĤ1ĭ%Ï|1qá96-90Ģ/ć,%x¿ÂĞÑ1|Àt1þB0·å¡0/(+ņōŒ1±÷1o;ŏ1^LO
lAĵ

6

%$1Đä^LOlüĤÜĄ2ÙĹŋâ»¬ŌĄ02Ĺoì-Ĉu°îŋ#>9p¬ł»Ō1Ŏĵ>66%ô--90¤ÛAį%

1:/YFld`AĽ"=-2ą-+98+1ďŊ,%>6,0%y1ĞÑ1|-ê5+9´k-90ĝ91,(%-ĚĮ+6"ė1þ

B0-(+9¬©ÞÍĕ/ÚľAĴ+%&%91-!+6"1:/|0"=-,%12ÉīðAÃ+&(%Ő1ī¼1÷1Ń,<
1¨A<+ïÎĬě

6";0«¬/ħĺ/,21:/ÞÍĕ/ĞÑ

YFld`2Ľà/(%-Ë6"÷}éęĿĳ±Ķ|1þè×ß±|tº1þè01¨A<+Æù

n6"

-Ŏŏ-03<%-$#'12    (*7:99;8=+ -$#' )5 =l[°wl¸»[,=öôôúõõ÷ðâñ
&=} ï²Ö{ (V¹ 3190)
   ðìi   }#+èçØg}iRØ¦g¥ãd,sñ0476 URLühttp://www.toyama-u.ac.jp/jp/Outline/access/= ½g~Õ×zç§lªAKN@

äµ

WQ ð}MÏñ
¾ÎË/±

¤Ê¡



P«`~ ðmåb$TëÁÉ¤Ê/ng}M¡ºñf/'ÁÉ·q,Â&

² È ð¹W}Mñ]e

BN6EN3N=FNKLÁÉeýþöó91<D:46¨²Ò ð}M¯ñ C68NµÓc¼£á/±Ðî¤ÊÑ¶

Þ\~_©

äµaÜ ðmåb

}M¡ºñjÔ´Í¯ ð¢}M°w¯ñÆl)6;J6*°-8L?4S

0D>á

G7Dl

ÐW¿~ ðXà}Mo~®íñ                   /4."6

!=öôôùõöûðâñOÌÖ­C68NµÓ/rY%µÓÇ!A4 ±ÀõBN5ÅðQRs 2.5 cm^¶ñéMµÓÇðÝv

Ãbñ *"
%



Ú,

ßÚ
òµÓÇ

Ö­

/

Ùç~ENIðMicrosoft word ¬Yëñ fbc8@jaist.ac.jp

ç~ENI(hÛZR|t!µÓÇRÄñ
MMuxyðÝÚYæ

|t!AHL;02;(f/(µÓ³&

%

Uipk³/*.

%

-öø-

ê

/

$"46,5

7

5Bh^nkGC qLDRuw88M 5,000 @ak 7,000 @8M 6,000 @ak 8,000 @
Rq 2,000 @ak 3,000 @
GC(nk 4H_h!bh!?TpKdYh8M1><R88M18M1RqAz=hk(Q.2/"
 FAX 4s$

#GC|pK16i'%#uwS3g&OJh6
s$

hH_:X 
H_KqLDR*0,+)-UZh!:X;7hry(9r4

/3$"eij  @

'%.!

923-1292 t[}{]lI 1-1
 E=vRfPRPR ojvRuwv ~N`0761-51-1681 FAX0761-51-1149
Q.2/fbc8@jaist.ac.jp
28(-&#)01

  *+67   xFVmUZP\hshinoha@eng.toyama-u.ac.jp   ~N`E=Pojhohsaka@jaist.ac.jp   W cUZP\shinono@eng.toyama-u.ac.jp

--


$/K 
 4E' 0  2 1=B<&><I:)  13%.!;"8AFC%.D!

                              
@G-+K#H 9J6(*5J,<?7