【手続補正書】
【提出日】平成16年10月19日(2004.10.19)
【手続補正１】
【補正対象書類名】明細書
【補正対象項目名】００７３
【補正方法】変更
【補正の内容】
【００７３】
公開されているSso7dのアミノ酸配列に基づき、Sso7dをコードする合成遺伝子を構築するうえで7個のオリゴヌクレオチドを使用した。オリゴヌクレオチドをアニーリングさせ、T4 DNAリガーゼを用いて連結させた。完全長の遺伝子を増幅させるため、2個の末端オリゴヌクレオチドをプライマーとして用い、連結した最終産物をPCRの鋳型として使用した。得られるPCR断片には、5’末端に唯一のEcoRI部位と、3’末端に唯一のBstXI部位が含まれるように設計した。Sso7dタンパク質をコードするほか、前記PCR断片はまた、Sso7dタンパク質のC末端に位置するGly-Gly-Val-Thr（配列番号：34）のアミノ酸配列を有するペプチドリンカーをコードする。Sso7dの合成遺伝子は、配列番号:1に示されるDNA配列を有し、配列番号:2に示されるアミノ酸配列を有するポリペプチドをコードする。
【手続補正２】
【補正対象書類名】明細書
【補正対象項目名】００７４
【補正方法】変更
【補正の内容】
【００７４】
Sso7dをコードする合成遺伝子を次に、Taqの最初の289アミノ酸がSso7dに置換される融合タンパク質を作製するために使用した。融合タンパク質を作製するため、以下のように、Sso7dをコードする断片を、Taqポリメラーゼをコードするプラスミドにサブクローニングした。手短に言えば、Sso7d合成遺伝子を含むDNA断片を制限酵素のEcoRIおよびBstXIで消化し、それからTaqをコードするプラスミドの対応部位にこれを連結した。結果として、Taqの最初の289アミノ酸をコードする領域がSso7d合成遺伝子に置換される。このプラスミド(pYW1)により、Gly-Gly-Val-Thr（配列番号：34）からなる合成リンカーを介してΔTaqのN末端に融合されたSso7dを含む単一ポリペプチドの発現が可能となる。融合タンパク質(Sso7d-ΔTaq)をコードするDNA配列およびそのタンパク質のアミノ酸配列はそれぞれ、配列番号:3および配列番号:4に示されている。
【手続補正３】
【補正対象書類名】明細書
【補正対象項目名】００７５
【補正方法】変更
【補正の内容】
【００７５】
Sso7d-Taq融合タンパク質の構築
Sso7d/完全長Taq融合タンパク質もまた構築した。手短に言えば、Taqポリメラーゼの最初の336アミノ酸をコードする1kbのPCR断片を、2個のプライマーを用いて作製した。5’プライマーによりPCR断片の5’末端にSpeI部位が導入され、3’プライマーはTaq遺伝子のヌクレオチド1008〜1026にハイブリダイズする。この断片をSpeIおよびBstXIで消化すると、Taqポリメラーゼの最初の289アミノ酸をコードする0.9kbの断片が放出される。この0.9kb断片をプラスミドpYW1にSpeI(リンカーをコードする領域に位置する)およびBstXI部位で連結した。得られるプラスミド(pYW2)により、Sso7d-ΔTaqと同様にして、Gly-Gly-Val-Thr（配列番号：34）からなるリンカーを介して完全長Taq DNAポリメラーゼのN末端に融合されたSso7dタンパク質を含む単一ポリペプチドの発現が可能となる。融合タンパク質Sso7d-TaqをコードするDNA配列およびそのタンパク質のアミノ酸配列はそれぞれ、配列番号:5および配列番号:6に示されている。
【手続補正４】
【補正対象書類名】明細書
【補正対象項目名】００７７
【補正方法】変更
【補正の内容】
【００７７】
上述のSso7d合成遺伝子をPCR増幅して、KpnI部位およびNheI部位をSso7d遺伝子に隣接するように導入するために、2個のプライマーを使用した。5’プライマーによってもまた、Sso7dタンパク質のN末端に、リンカーとして機能する6個の付加アミノ酸(Gly-Thr-Gly-Gly-Gly-Gly；配列番号：35)が導入された。KpnIおよびNheIで消化後、PCR断片をpPFKSに対応部位で連結した。得られるプラスミド(pPFS)により、ペプチドリンカー(Gly-Thr-Gly-Gly-Gly-Gly；配列番号：35)を介してPfuポリメラーゼのC末端に融合されたSso7dタンパク質を含む単一ポリペプチドの発現が可能となる。融合タンパク質(Pfu-Sso7d)をコードするDNA配列およびその融合タンパク質のアミノ酸配列はそれぞれ、配列番号:7および配列番号:8に示されている。
【手続補正５】
【補正対象書類名】明細書
【補正対象項目名】００７９
【補正方法】変更
【補正の内容】
【００７９】
PL-ΔTaq融合タンパク質の構築
5番目の融合タンパク質は、14個のリジンと2個のアルギニンからなるペプチドがΔTaqのN末端に連結されている。ポリリジン(PL)-ΔTaq融合タンパク質を作製するため、2個の67ヌクレオチド(nt)のオリゴヌクレオチドをアニーリングさせて、EcoRI部位と適合する5’突出末端、およびSpeI部位と適合する3’突出末端を有する二本鎖DNA断片を形成させた。このDNA断片は、以下の組成のリジンが豊富なペプチドをコードする: 
2005510235.tif 000001
（配列番号：36）。このペプチド中のリジンおよびアルギニンの数は、Sso7d中のものと同じである。このDNA断片を、EcoRIおよびSpeIで予め消化されたpYW1に、Sso7dをコードする領域と置換されるように連結した。得られるプラスミド(pLST)により、ΔTaqのN末端に融合されたリジンが豊富なペプチドを含む単一ポリペプチドが発現される。融合タンパク質(PL-ΔTaq)をコードするDNA配列およびそのタンパク質のアミノ酸配列はそれぞれ、配列番号:11および配列番号:12に示されている。
【手続補正６】
【補正対象書類名】明細書
【補正対象項目名】０１１０
【補正方法】変更
【補正の内容】
【０１１０】
実施例8. Sso7d融合ポリメラーゼはサイクルシークエンシングに有利である
DNA配列決定法に適した改良型ポリメラーゼをコードするプラスミド・クローンを構築し、そのタンパク質産物を精製した。第一の酵素はSso7d-ΔTaq(Y) (変異を太字体で表示した配列番号:30および配列番号:31)であり、これは配列番号:31の表示位置で「F」残基の代わりに「Y」残基に置換されるように、TaborおよびRichardson (米国特許第5,614,365号)の方法により改変されていることを除いて、酵素Sso7d-ΔTaqと同じである。第二の酵素はSso7d-ΔTaq(E5;Y) (変異を太字体で表示した配列番号:32および配列番号:33) であり、これはTaborおよびRichardsonの方法により改変され、また5’から3’のヌクレアーゼドメインを不活性化する点突然変異を含むことを除いて、Sso7d-Taqと同じである。
【手続補正７】
【補正対象書類名】明細書
【補正対象項目名】０１１５
【補正方法】変更
【補正の内容】
【０１１５】
配列表
配列番号:1 合成Sso7d遺伝子
2005510235.tif 000002
配列番号:2 Sso7dのアミノ酸配列
2005510235.tif 000003
配列番号:3 Sso7d-ΔTaq融合タンパク質をコードするDNA配列
2005510235.tif 000004
2005510235.tif 000005
配列番号:4 Sso7d-ΔTaq融合タンパク質のアミノ酸配列
2005510235.tif 000006
配列番号:5 Sso7d-Taq融合タンパク質をコードするDNA配列
2005510235.tif 000007
2005510235.tif 000008
配列番号:6 Sso7d-Taq融合タンパク質のアミノ酸配列
2005510235.tif 000009
配列番号:7 Pfu-Sso7d融合タンパク質をコードするDNA配列
2005510235.tif 000010
2005510235.tif 000011
配列番号:8 Pfu-Sso7d融合タンパク質のアミノ酸配列
2005510235.tif 000012
配列番号:9 Sac7d-ΔTaq融合タンパク質をコードするDNA配列
2005510235.tif 000013
2005510235.tif 000014
配列番号:10 Sac7d-ΔTaq融合タンパク質のアミノ酸配列
2005510235.tif 000015
配列番号:11 PL-ΔTaq融合タンパク質をコードするDNA配列
2005510235.tif 000016
配列番号:12 PL-ΔTaq融合タンパク質のアミノ酸配列
2005510235.tif 000017
配列番号:13 プライマー L71F
2005510235.tif 000018
配列番号:14 プライマー L71R
2005510235.tif 000019
配列番号:15 プライマー L18015F
2005510235.tif 000020
配列番号:16 プライマー L23474R
2005510235.tif 000021
配列番号:17 プライマー L18015F
2005510235.tif 000022
配列番号:18 プライマー L29930R
2005510235.tif 000023
配列番号:19 プライマー L30350F
2005510235.tif 000024
配列番号:20 プライマー L35121R
2005510235.tif 000025
配列番号:21 プライマー L2089F
2005510235.tif 000026
配列番号:22 プライマー L7112R
2005510235.tif 000027
配列番号:23 プライマー L30350F
2005510235.tif 000028
配列番号:24 プライマー L40547R
2005510235.tif 000029
配列番号:25 プライマー H-Amelo-Y
2005510235.tif 000030
配列番号:26 プライマー H-Amelo-YR
2005510235.tif 000031
配列番号:27 ヒトβ-グロビン プライマー 536F
2005510235.tif 000032
配列番号:28 ヒトβ-グロビン プライマー 536R
2005510235.tif 000033
配列番号:29 ヒトβ-グロビン プライマー 1408R
2005510235.tif 000034
配列番号:30 Sso7d-ΔTaq(Y)タンパク質をコードするDNA配列
2005510235.tif 000035
配列番号:31 Sso7d-ΔTaq(Y)タンパク質のアミノ酸配列
2005510235.tif 000036
配列番号:32 Sso7d-ΔTaq(E5)(Y)タンパク質をコードするDNA配列
2005510235.tif 000037
2005510235.tif 000038
配列番号:33 Sso7d-ΔTaq(E5)(Y)タンパク質のアミノ酸配列
2005510235.tif 000039
【配列表】
SEQUENCE LISTING                               
<110> MJ Bioworks Incorporated
<120> Methods of Using Improved Polymerases
<130> TO-A0425
<140> JP   2003-547584
<141> 2002-11-27
<150> US   60/333,966
<151> 2001-11-28
<160> 36                            
<170> PatentIn Ver. 2.1
<210> 1
<211> 189
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:synthetic
Sulfolobus solfataricus Sso7d gene
<400> 1
gcaaccgtaa agttcaagta caaaggcgaa gaaaaagagg tagacatctc caagatcaag 60    
aaagtatggc gtgtgggcaa gatgatctcc ttcacctacg acgagggcgg tggcaagacc 120   
ggccgtggtg cggtaagcga aaaggacgcg ccgaaggagc tgctgcagat gctggagaag 180   
cagaaaaag                                                         189   
<210> 2
<211> 63
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sso7d
<400> 2
Ala Thr Val Lys Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp Ile  
1               5                  10                  15    
Ser Lys Ile Lys Lys Val Trp Arg Val Gly Lys Met Ile Ser Phe Thr  
20                  25                  30        
Tyr Asp Glu Gly Gly Gly Lys Thr Gly Arg Gly Ala Val Ser Glu Lys  
35                  40                  45            
Asp Ala Pro Lys Glu Leu Leu Gln Met Leu Glu Lys Gln Lys Lys      
50                  55                  60            
<210> 3
<211> 1899
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sso7d-deltaTaq
fusion protein
<400> 3
atgattacga attcgagcgc aaccgtaaag ttcaagtaca aaggcgaaga aaaagaggta 60    
gacatctcca agatcaagaa agtatggcgt gtgggcaaga tgatctcctt cacctacgac 120   
gagggcggtg gcaagaccgg ccgtggtgcg gtaagcgaaa aggacgcgcc gaaggagctg 180   
ctgcagatgc tggagaagca gaaaaagggc ggcggtgtca ctagtcccaa ggccctggag 240   
gaggccccct ggcccccgcc ggaaggggcc ttcgtgggct ttgtgctttc ccgcaaggag 300   
cccatgtggg ccgatcttct ggccctggcc gccgccaggg ggggccgggt ccaccgggcc 360   
cccgagcctt ataaagccct cagggacctg aaggaggcgc gggggcttct cgccaaagac 420   
ctgagcgttc tggccctgag ggaaggcctt ggcctcccgc ccggcgacga ccccatgctc 480   
ctcgcctacc tcctggaccc ttccaacacc acccccgagg gggtggcccg gcgctacggc 540   
ggggagtgga cggaggaggc gggggagcgg gccgcccttt ccgagaggct cttcgccaac 600   
ctgtggggga ggcttgaggg ggaggagagg ctcctttggc tttaccggga ggtggagagg 660   
cccctttccg ctgtcctggc ccacatggag gccacggggg tgcgcctgga cgtggcctat 720   
ctcagggcct tgtccctgga ggtggccgag gagatcgccc gcctcgaggc cgaggtcttc 780   
cgcctggccg gccacccctt caacctcaac tcccgggacc agctggaaag ggtcctcttt 840   
gacgagctag ggcttcccgc catcggcaag acggagaaga ccggcaagcg ctccaccagc 900   
gccgccgtcc tggaggccct ccgcgaggcc caccccatcg tggagaagat cctgcagtac 960   
cgggagctca ccaagctgaa gagcacctac attgacccct tgccggacct catccacccc 1020  
aggacgggcc gcctccacac ccgcttcaac cagacggcca cggccacggg caggctaagt 1080  
agctccgatc ccaacctcca gaacatcccc gtccgcaccc cgcttgggca gaggatccgc 1140  
cgggccttca tcgccgagga ggggtggcta ttggtggccc tggactatag ccagatagag 1200  
ctcagggtgc tggcccacct ctccggcgac gagaacctga tccgggtctt ccaggagggg 1260  
cgggacatcc acacggagac cgccagctgg atgttcggcg tcccccggga ggccgtggac 1320  
cccctgatgc gccgggcggc caagaccatc aacttcgggg tcctctacgg catgtcggcc 1380  
caccgcctct cccaggagct agccatccct tacgaggagg cccaggcctt cattgagcgc 1440  
tactttcaga gcttccccaa ggtgcgggcc tggattgaga agaccctgga ggagggcagg 1500  
aggcgggggt acgtggagac cctcttcggc cgccgccgct acgtgccaga cctagaggcc 1560  
cgggtgaaga gcgtgcggga ggcggccgag cgcatggcct tcaacatgcc cgtccagggc 1620  
accgccgccg acctcatgaa gctggctatg gtgaagctct tccccaggct ggaggaaatg 1680  
ggggccagga tgctccttca ggtccacgac gagctggtcc tcgaggcccc aaaagagagg 1740  
gcggaggccg tggcccggct ggccaaggag gtcatggagg gggtgtatcc cctggccgtg 1800  
cccctggagg tggaggtggg gataggggag gactggctct ccgccaagga gggcattgat 1860  
ggccgcggcg gaggcgggca tcatcatcat catcattaa                        1899  
<210> 4
<211> 632
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sso7d-deltaTaq
fusion protein
<400> 4
Met Ile Thr Asn Ser Ser Ala Thr Val Lys Phe Lys Tyr Lys Gly Glu  
1               5                  10                  15    
Glu Lys Glu Val Asp Ile Ser Lys Ile Lys Lys Val Trp Arg Val Gly  
20                  25                  30        
Lys Met Ile Ser Phe Thr Tyr Asp Glu Gly Gly Gly Lys Thr Gly Arg  
35                  40                  45            
Gly Ala Val Ser Glu Lys Asp Ala Pro Lys Glu Leu Leu Gln Met Leu  
50                  55                  60                
Glu Lys Gln Lys Lys Gly Gly Gly Val Thr Ser Pro Lys Ala Leu Glu  
65                  70                  75                  80
Glu Ala Pro Trp Pro Pro Pro Glu Gly Ala Phe Val Gly Phe Val Leu  
85                  90                  95    
Ser Arg Lys Glu Pro Met Trp Ala Asp Leu Leu Ala Leu Ala Ala Ala  
100                 105                 110        
Arg Gly Gly Arg Val His Arg Ala Pro Glu Pro Tyr Lys Ala Leu Arg  
115                 120                 125            
Asp Leu Lys Glu Ala Arg Gly Leu Leu Ala Lys Asp Leu Ser Val Leu  
130                 135                 140                
Ala Leu Arg Glu Gly Leu Gly Leu Pro Pro Gly Asp Asp Pro Met Leu  
145                 150                 155                 160
Leu Ala Tyr Leu Leu Asp Pro Ser Asn Thr Thr Pro Glu Gly Val Ala  
165                 170                 175    
Arg Arg Tyr Gly Gly Glu Trp Thr Glu Glu Ala Gly Glu Arg Ala Ala  
180                 185                 190        
Leu Ser Glu Arg Leu Phe Ala Asn Leu Trp Gly Arg Leu Glu Gly Glu  
195                 200                 205            
Glu Arg Leu Leu Trp Leu Tyr Arg Glu Val Glu Arg Pro Leu Ser Ala  
210                 215                 220                
Val Leu Ala His Met Glu Ala Thr Gly Val Arg Leu Asp Val Ala Tyr  
225                 230                 235                 240
Leu Arg Ala Leu Ser Leu Glu Val Ala Glu Glu Ile Ala Arg Leu Glu  
245                 250                 255    
Ala Glu Val Phe Arg Leu Ala Gly His Pro Phe Asn Leu Asn Ser Arg  
260                 265                 270        
Asp Gln Leu Glu Arg Val Leu Phe Asp Glu Leu Gly Leu Pro Ala Ile  
275                 280                 285            
Gly Lys Thr Glu Lys Thr Gly Lys Arg Ser Thr Ser Ala Ala Val Leu  
290                 295                 300                
Glu Ala Leu Arg Glu Ala His Pro Ile Val Glu Lys Ile Leu Gln Tyr  
305                 310                 315                 320
Arg Glu Leu Thr Lys Leu Lys Ser Thr Tyr Ile Asp Pro Leu Pro Asp  
325                 330                 335    
Leu Ile His Pro Arg Thr Gly Arg Leu His Thr Arg Phe Asn Gln Thr  
340                 345                 350        
Ala Thr Ala Thr Gly Arg Leu Ser Ser Ser Asp Pro Asn Leu Gln Asn  
355                 360                 365            
Ile Pro Val Arg Thr Pro Leu Gly Gln Arg Ile Arg Arg Ala Phe Ile  
370                 375                 380                
Ala Glu Glu Gly Trp Leu Leu Val Ala Leu Asp Tyr Ser Gln Ile Glu  
385                 390                 395                 400
Leu Arg Val Leu Ala His Leu Ser Gly Asp Glu Asn Leu Ile Arg Val  
405                 410                 415    
Phe Gln Glu Gly Arg Asp Ile His Thr Glu Thr Ala Ser Trp Met Phe  
420                 425                 430        
Gly Val Pro Arg Glu Ala Val Asp Pro Leu Met Arg Arg Ala Ala Lys  
435                 440                 445            
Thr Ile Asn Phe Gly Val Leu Tyr Gly Met Ser Ala His Arg Leu Ser  
450                 455                 460                
Gln Glu Leu Ala Ile Pro Tyr Glu Glu Ala Gln Ala Phe Ile Glu Arg  
465                 470                 475                 480
Tyr Phe Gln Ser Phe Pro Lys Val Arg Ala Trp Ile Glu Lys Thr Leu  
485                 490                 495    
Glu Glu Gly Arg Arg Arg Gly Tyr Val Glu Thr Leu Phe Gly Arg Arg  
500                 505                 510        
Arg Tyr Val Pro Asp Leu Glu Ala Arg Val Lys Ser Val Arg Glu Ala  
515                 520                 525            
Ala Glu Arg Met Ala Phe Asn Met Pro Val Gln Gly Thr Ala Ala Asp  
530                 535                 540                
Leu Met Lys Leu Ala Met Val Lys Leu Phe Pro Arg Leu Glu Glu Met  
545                 550                 555                 560
Gly Ala Arg Met Leu Leu Gln Val His Asp Glu Leu Val Leu Glu Ala  
565                 570                 575    
Pro Lys Glu Arg Ala Glu Ala Val Ala Arg Leu Ala Lys Glu Val Met  
580                 585                 590        
Glu Gly Val Tyr Pro Leu Ala Val Pro Leu Glu Val Glu Val Gly Ile  
595                 600                 605            
Gly Glu Asp Trp Leu Ser Ala Lys Glu Gly Ile Asp Gly Arg Gly Gly  
610                 615                 620                
Gly Gly His His His His His His                                  
625                 630        
<210> 5
<211> 2763
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sso7d-Taq
fusion protein, Sso7d/full-length Taq fusion
protein
<400> 5
atgattacga attcgagcgc aaccgtaaag ttcaagtaca aaggcgaaga aaaagaggta 60    
gacatctcca agatcaagaa agtatggcgt gtgggcaaga tgatctcctt cacctacgac 120   
gagggcggtg gcaagaccgg ccgtggtgcg gtaagcgaaa aggacgcgcc gaaggagctg 180   
ctgcagatgc tggagaagca gaaaaagggc ggcggtgtca ctagtgggat gctgcccctc 240   
tttgagccca agggccgggt cctcctggtg gacggccacc acctggccta ccgcaccttc 300   
cacgccctga agggcctcac caccagccgg ggggagccgg tgcaggcggt ctacggcttc 360   
gccaagagcc tcctcaaggc cctcaaggag gacggggacg cggtgatcgt ggtctttgac 420   
gccaaggccc cctccttccg ccacgaggcc tacggggggt acaaggcggg ccgggccccc 480   
acgccagagg actttccccg gcaactcgcc ctcatcaagg agctggtgga cctcctgggg 540   
ctggcgcgcc tcgaggtccc gggctacgag gcggacgacg tcctggccag cctggccaag 600   
aaggcggaaa aggagggcta cgaggtccgc atcctcaccg ccgacaaaga cctttaccag 660   
ctcctttccg accgcatcca cgtcctccac cccgaggggt acctcatcac cccggcctgg 720   
ctttgggaaa agtacggcct gaggcccgac cagtgggccg actaccgggc cctgaccggg 780   
gacgagtccg acaaccttcc cggggtcaag ggcatcgggg agaagacggc gaggaagctt 840   
ctggaggagt gggggagcct ggaagccctc ctcaagaacc tggaccggct gaagcccgcc 900   
atccgggaga agatcctggc ccacatggac gatctgaagc tctcctggga cctggccaag 960   
gtgcgcaccg acctgcccct ggaggtggac ttcgccaaaa ggcgggagcc cgaccgggag 1020  
aggcttaggg cctttctgga gaggcttgag tttggcagcc tcctccacga gttcggcctt 1080  
ctggaaagcc ccaaggccct ggaggaggcc ccctggcccc cgccggaagg ggccttcgtg 1140  
ggctttgtgc tttcccgcaa ggagcccatg tgggccgatc ttctggccct ggccgccgcc 1200  
agggggggcc gggtccaccg ggcccccgag ccttataaag ccctcaggga cctgaaggag 1260  
gcgcgggggc ttctcgccaa agacctgagc gttctggccc tgagggaagg ccttggcctc 1320  
ccgcccggcg acgaccccat gctcctcgcc tacctcctgg acccttccaa caccaccccc 1380  
gagggggtgg cccggcgcta cggcggggag tggacggagg aggcggggga gcgggccgcc 1440  
ctttccgaga ggctcttcgc caacctgtgg gggaggcttg agggggagga gaggctcctt 1500  
tggctttacc gggaggtgga gaggcccctt tccgctgtcc tggcccacat ggaggccacg 1560  
ggggtgcgcc tggacgtggc ctatctcagg gccttgtccc tggaggtggc cgaggagatc 1620  
gcccgcctcg aggccgaggt cttccgcctg gccggccacc ccttcaacct caactcccgg 1680  
gaccagctgg aaagggtcct ctttgacgag ctagggcttc ccgccatcgg caagacggag 1740  
aagaccggca agcgctccac cagcgccgcc gtcctggagg ccctccgcga ggcccacccc 1800  
atcgtggaga agatcctgca gtaccgggag ctcaccaagc tgaagagcac ctacattgac 1860  
cccttgccgg acctcatcca ccccaggacg ggccgcctcc acacccgctt caaccagacg 1920  
gccacggcca cgggcaggct aagtagctcc gatcccaacc tccagaacat ccccgtccgc 1980  
accccgcttg ggcagaggat ccgccgggcc ttcatcgccg aggaggggtg gctattggtg 2040  
gccctggact atagccagat agagctcagg gtgctggccc acctctccgg cgacgagaac 2100  
ctgatccggg tcttccagga ggggcgggac atccacacgg agaccgccag ctggatgttc 2160  
ggcgtccccc gggaggccgt ggaccccctg atgcgccggg cggccaagac catcaacttc 2220  
ggggtcctct acggcatgtc ggcccaccgc ctctcccagg agctagccat cccttacgag 2280  
gaggcccagg ccttcattga gcgctacttt cagagcttcc ccaaggtgcg ggcctggatt 2340  
gagaagaccc tggaggaggg caggaggcgg gggtacgtgg agaccctctt cggccgccgc 2400  
cgctacgtgc cagacctaga ggcccgggtg aagagcgtgc gggaggcggc cgagcgcatg 2460  
gccttcaaca tgcccgtcca gggcaccgcc gccgacctca tgaagctggc tatggtgaag 2520  
ctcttcccca ggctggagga aatgggggcc aggatgctcc ttcaggtcca cgacgagctg 2580  
gtcctcgagg ccccaaaaga gagggcggag gccgtggccc ggctggccaa ggaggtcatg 2640  
gagggggtgt atcccctggc cgtgcccctg gaggtggagg tggggatagg ggaggactgg 2700  
ctctccgcca aggagggcat tgatggccgc ggcggaggcg ggcatcatca tcatcatcat 2760  
taa                                                               2763  
<210> 6
<211> 920
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sso7d-Taq
fusion protein, Sso7d/full-length Taq fusion
protein
<400> 6
Met Ile Thr Asn Ser Ser Ala Thr Val Lys Phe Lys Tyr Lys Gly Glu  
1               5                  10                  15    
Glu Lys Glu Val Asp Ile Ser Lys Ile Lys Lys Val Trp Arg Val Gly  
20                  25                  30        
Lys Met Ile Ser Phe Thr Tyr Asp Glu Gly Gly Gly Lys Thr Gly Arg  
35                  40                  45            
Gly Ala Val Ser Glu Lys Asp Ala Pro Lys Glu Leu Leu Gln Met Leu  
50                  55                  60                
Glu Lys Gln Lys Lys Gly Gly Gly Val Thr Ser Gly Met Leu Pro Leu  
65                  70                  75                  80
Phe Glu Pro Lys Gly Arg Val Leu Leu Val Asp Gly His His Leu Ala  
85                  90                  95    
Tyr Arg Thr Phe His Ala Leu Lys Gly Leu Thr Thr Ser Arg Gly Glu  
100                 105                 110        
Pro Val Gln Ala Val Tyr Gly Phe Ala Lys Ser Leu Leu Lys Ala Leu  
115                 120                 125            
Lys Glu Asp Gly Asp Ala Val Ile Val Val Phe Asp Ala Lys Ala Pro  
130                 135                 140                
Ser Phe Arg His Glu Ala Tyr Gly Gly Tyr Lys Ala Gly Arg Ala Pro  
145                 150                 155                 160
Thr Pro Glu Asp Phe Pro Arg Gln Leu Ala Leu Ile Lys Glu Leu Val  
165                 170                 175    
Asp Leu Leu Gly Leu Ala Arg Leu Glu Val Pro Gly Tyr Glu Ala Asp  
180                 185                 190        
Asp Val Leu Ala Ser Leu Ala Lys Lys Ala Glu Lys Glu Gly Tyr Glu  
195                 200                 205            
Val Arg Ile Leu Thr Ala Asp Lys Asp Leu Tyr Gln Leu Leu Ser Asp  
210                 215                 220                
Arg Ile His Val Leu His Pro Glu Gly Tyr Leu Ile Thr Pro Ala Trp  
225                 230                 235                 240
Leu Trp Glu Lys Tyr Gly Leu Arg Pro Asp Gln Trp Ala Asp Tyr Arg  
245                 250                 255    
Ala Leu Thr Gly Asp Glu Ser Asp Asn Leu Pro Gly Val Lys Gly Ile  
260                 265                 270        
Gly Glu Lys Thr Ala Arg Lys Leu Leu Glu Glu Trp Gly Ser Leu Glu  
275                 280                 285            
Ala Leu Leu Lys Asn Leu Asp Arg Leu Lys Pro Ala Ile Arg Glu Lys  
290                 295                 300                
Ile Leu Ala His Met Asp Asp Leu Lys Leu Ser Trp Asp Leu Ala Lys  
305                 310                 315                 320
Val Arg Thr Asp Leu Pro Leu Glu Val Asp Phe Ala Lys Arg Arg Glu  
325                 330                 335    
Pro Asp Arg Glu Arg Leu Arg Ala Phe Leu Glu Arg Leu Glu Phe Gly  
340                 345                 350        
Ser Leu Leu His Glu Phe Gly Leu Leu Glu Ser Pro Lys Ala Leu Glu  
355                 360                 365            
Glu Ala Pro Trp Pro Pro Pro Glu Gly Ala Phe Val Gly Phe Val Leu  
370                 375                 380                
Ser Arg Lys Glu Pro Met Trp Ala Asp Leu Leu Ala Leu Ala Ala Ala  
385                 390                 395                 400
Arg Gly Gly Arg Val His Arg Ala Pro Glu Pro Tyr Lys Ala Leu Arg  
405                 410                 415    
Asp Leu Lys Glu Ala Arg Gly Leu Leu Ala Lys Asp Leu Ser Val Leu  
420                 425                 430        
Ala Leu Arg Glu Gly Leu Gly Leu Pro Pro Gly Asp Asp Pro Met Leu  
435                 440                 445            
Leu Ala Tyr Leu Leu Asp Pro Ser Asn Thr Thr Pro Glu Gly Val Ala  
450                 455                 460                
Arg Arg Tyr Gly Gly Glu Trp Thr Glu Glu Ala Gly Glu Arg Ala Ala  
465                 470                 475                 480
Leu Ser Glu Arg Leu Phe Ala Asn Leu Trp Gly Arg Leu Glu Gly Glu  
485                 490                 495    
Glu Arg Leu Leu Trp Leu Tyr Arg Glu Val Glu Arg Pro Leu Ser Ala  
500                 505                 510        
Val Leu Ala His Met Glu Ala Thr Gly Val Arg Leu Asp Val Ala Tyr  
515                 520                 525            
Leu Arg Ala Leu Ser Leu Glu Val Ala Glu Glu Ile Ala Arg Leu Glu  
530                 535                 540                
Ala Glu Val Phe Arg Leu Ala Gly His Pro Phe Asn Leu Asn Ser Arg  
545                 550                 555                 560
Asp Gln Leu Glu Arg Val Leu Phe Asp Glu Leu Gly Leu Pro Ala Ile  
565                 570                 575    
Gly Lys Thr Glu Lys Thr Gly Lys Arg Ser Thr Ser Ala Ala Val Leu  
580                 585                 590        
Glu Ala Leu Arg Glu Ala His Pro Ile Val Glu Lys Ile Leu Gln Tyr  
595                 600                 605            
Arg Glu Leu Thr Lys Leu Lys Ser Thr Tyr Ile Asp Pro Leu Pro Asp  
610                 615                 620                
Leu Ile His Pro Arg Thr Gly Arg Leu His Thr Arg Phe Asn Gln Thr  
625                 630                 635                 640
Ala Thr Ala Thr Gly Arg Leu Ser Ser Ser Asp Pro Asn Leu Gln Asn  
645                 650                 655    
Ile Pro Val Arg Thr Pro Leu Gly Gln Arg Ile Arg Arg Ala Phe Ile  
660                 665                 670        
Ala Glu Glu Gly Trp Leu Leu Val Ala Leu Asp Tyr Ser Gln Ile Glu  
675                 680                 685            
Leu Arg Val Leu Ala His Leu Ser Gly Asp Glu Asn Leu Ile Arg Val  
690                 695                 700                
Phe Gln Glu Gly Arg Asp Ile His Thr Glu Thr Ala Ser Trp Met Phe  
705                 710                 715                 720
Gly Val Pro Arg Glu Ala Val Asp Pro Leu Met Arg Arg Ala Ala Lys  
725                 730                 735    
Thr Ile Asn Phe Gly Val Leu Tyr Gly Met Ser Ala His Arg Leu Ser  
740                 745                 750        
Gln Glu Leu Ala Ile Pro Tyr Glu Glu Ala Gln Ala Phe Ile Glu Arg  
755                 760                 765            
Tyr Phe Gln Ser Phe Pro Lys Val Arg Ala Trp Ile Glu Lys Thr Leu  
770                 775                 780                
Glu Glu Gly Arg Arg Arg Gly Tyr Val Glu Thr Leu Phe Gly Arg Arg  
785                 790                 795                 800
Arg Tyr Val Pro Asp Leu Glu Ala Arg Val Lys Ser Val Arg Glu Ala  
805                 810                 815    
Ala Glu Arg Met Ala Phe Asn Met Pro Val Gln Gly Thr Ala Ala Asp  
820                 825                 830        
Leu Met Lys Leu Ala Met Val Lys Leu Phe Pro Arg Leu Glu Glu Met  
835                 840                 845            
Gly Ala Arg Met Leu Leu Gln Val His Asp Glu Leu Val Leu Glu Ala  
850                 855                 860                
Pro Lys Glu Arg Ala Glu Ala Val Ala Arg Leu Ala Lys Glu Val Met  
865                 870                 875                 880
Glu Gly Val Tyr Pro Leu Ala Val Pro Leu Glu Val Glu Val Gly Ile  
885                 890                 895    
Gly Glu Asp Trp Leu Ser Ala Lys Glu Gly Ile Asp Gly Arg Gly Gly  
900                 905                 910        
Gly Gly His His His His His His                                  
915                 920
<210> 7
<211> 2535
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Pfu-Sso7d
fusion protein
<400> 7
atgattttag atgtggatta cataactgaa gaaggaaaac ctgttattag gctattcaaa 60    
aaagagaacg gaaaatttaa gatagagcat gatagaactt ttagaccata catttacgct 120   
cttctcaggg atgattcaaa gattgaagaa gttaagaaaa taacggggga aaggcatgga 180   
aagattgtga gaattgttga tgtagagaag gttgagaaaa agtttctcgg caagcctatt 240   
accgtgtgga aactttattt ggaacatccc caagatgttc ccactattag agaaaaagtt 300   
agagaacatc cagcagttgt ggacatcttc gaatacgata ttccatttgc aaagagatac 360   
ctcatcgaca aaggcctaat accaatggag ggggaagaag agctaaagat tcttgccttc 420   
gatatagaaa ccctctatca cgaaggagaa gagtttggaa aaggcccaat tataatgatt 480   
agttatgcag atgaaaatga agcaaaggtg attacttgga aaaacataga tcttccatac 540   
gttgaggttg tatcaagcga gagagagatg ataaagagat ttctcaggat tatcagggag 600   
aaggatcctg acattatagt tacttataat ggagactcat tcgacttccc atatttagcg 660   
aaaagggcag aaaaacttgg gattaaatta accattggaa gagatggaag cgagcccaag 720   
atgcagagaa taggcgatat gacggctgta gaagtcaagg gaagaataca tttcgacttg 780   
tatcatgtaa taacaaggac aataaatctc ccaacataca cactagaggc tgtatatgaa 840   
gcaatttttg gaaagccaaa ggagaaggta tacgccgacg agatagcaaa agcctgggaa 900   
agtggagaga accttgagag agttgccaaa tactcgatgg aagatgcaaa ggcaacttat 960   
gaactcggga aagaattcct tccaatggaa attcagcttt caagattagt tggacaacct 1020  
ttatgggatg tttcaaggtc aagcacaggg aaccttgtag agtggttctt acttaggaaa 1080  
gcctacgaaa gaaacgaagt agctccaaac aagccaagtg aagaggagta tcaaagaagg 1140  
ctcagggaga gctacacagg tggattcgtt aaagagccag aaaaggggtt gtgggaaaac 1200  
atagtatacc tagattttag agccctatat ccctcgatta taattaccca caatgtttct 1260  
cccgatactc taaatcttga gggatgcaag aactatgata tcgctcctca agtaggccac 1320  
aagttctgca aggacatccc tggttttata ccaagtctct tgggacattt gttagaggaa 1380  
agacaaaaga ttaagacaaa aatgaaggaa actcaagatc ctatagaaaa aatactcctt 1440  
gactatagac aaaaagcgat aaaactctta gcaaattctt tctacggata ttatggctat 1500  
gcaaaagcaa gatggtactg taaggagtgt gctgagagcg ttactgcctg gggaagaaag 1560  
tacatcgagt tagtatggaa ggagctcgaa gaaaagtttg gatttaaagt cctctacatt 1620  
gacactgatg gtctctatgc aactatccca ggaggagaaa gtgaggaaat aaagaaaaag 1680  
gctctagaat ttgtaaaata cataaattca aagctccctg gactgctaga gcttgaatat 1740  
gaagggtttt ataagagggg attcttcgtt acgaagaaga ggtatgcagt aatagatgaa 1800  
gaaggaaaag tcattactcg tggtttagag atagttagga gagattggag tgaaattgca 1860  
aaagaaactc aagctagagt tttggagaca atactaaaac acggagatgt tgaagaagct 1920  
gtgagaatag taaaagaagt aatacaaaag cttgccaatt atgaaattcc accagagaag 1980  
ctcgcaatat atgagcagat aacaagacca ttacatgagt ataaggcgat aggtcctcac 2040  
gtagctgttg caaagaaact agctgctaaa ggagttaaaa taaagccagg aatggtaatt 2100  
ggatacatag tacttagagg cgatggtcca attagcaata gggcaattct agctgaggaa 2160  
tacgatccca aaaagcacaa gtatgacgca gaatattaca ttgagaacca ggttcttcca 2220  
gcggtactta ggatattgga gggatttgga tacagaaagg aagacctcag ataccaaaag 2280  
acaagacaag tcggcctaac ttcctggctt aacattaaaa aatccggtac cggcggtggc 2340  
ggtgcaaccg taaagttcaa gtacaaaggc gaagaaaaag aggtagacat ctccaagatc 2400  
aagaaagtat ggcgtgtggg caagatgatc tccttcacct acgacgaggg cggtggcaag 2460  
accggccgtg gtgcggtaag cgaaaaggac gcgccgaagg agctgctgca gatgctggag 2520  
aagcagaaaa agtga                                                  2535  
<210> 8
<211> 844
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Pfu-Sso7d
fusion protein
<400> 8
Met Ile Leu Asp Val Asp Tyr Ile Thr Glu Glu Gly Lys Pro Val Ile  
1               5                  10                  15    
Arg Leu Phe Lys Lys Glu Asn Gly Lys Phe Lys Ile Glu His Asp Arg  
20                  25                  30        
Thr Phe Arg Pro Tyr Ile Tyr Ala Leu Leu Arg Asp Asp Ser Lys Ile  
35                  40                  45            
Glu Glu Val Lys Lys Ile Thr Gly Glu Arg His Gly Lys Ile Val Arg  
50                  55                  60                
Ile Val Asp Val Glu Lys Val Glu Lys Lys Phe Leu Gly Lys Pro Ile  
65                  70                  75                  80
Thr Val Trp Lys Leu Tyr Leu Glu His Pro Gln Asp Val Pro Thr Ile  
85                  90                  95    
Arg Glu Lys Val Arg Glu His Pro Ala Val Val Asp Ile Phe Glu Tyr  
100                 105                 110        
Asp Ile Pro Phe Ala Lys Arg Tyr Leu Ile Asp Lys Gly Leu Ile Pro  
115                 120                 125            
Met Glu Gly Glu Glu Glu Leu Lys Ile Leu Ala Phe Asp Ile Glu Thr  
130                 135                 140                
Leu Tyr His Glu Gly Glu Glu Phe Gly Lys Gly Pro Ile Ile Met Ile  
145                 150                 155                 160
Ser Tyr Ala Asp Glu Asn Glu Ala Lys Val Ile Thr Trp Lys Asn Ile  
165                 170                 175    
Asp Leu Pro Tyr Val Glu Val Val Ser Ser Glu Arg Glu Met Ile Lys  
180                 185                 190        
Arg Phe Leu Arg Ile Ile Arg Glu Lys Asp Pro Asp Ile Ile Val Thr  
195                 200                 205            
Tyr Asn Gly Asp Ser Phe Asp Phe Pro Tyr Leu Ala Lys Arg Ala Glu  
210                 215                 220                
Lys Leu Gly Ile Lys Leu Thr Ile Gly Arg Asp Gly Ser Glu Pro Lys  
225                 230                 235                 240
Met Gln Arg Ile Gly Asp Met Thr Ala Val Glu Val Lys Gly Arg Ile  
245                 250                 255    
His Phe Asp Leu Tyr His Val Ile Thr Arg Thr Ile Asn Leu Pro Thr  
260                 265                 270        
Tyr Thr Leu Glu Ala Val Tyr Glu Ala Ile Phe Gly Lys Pro Lys Glu  
275                 280                 285            
Lys Val Tyr Ala Asp Glu Ile Ala Lys Ala Trp Glu Ser Gly Glu Asn  
290                 295                 300                
Leu Glu Arg Val Ala Lys Tyr Ser Met Glu Asp Ala Lys Ala Thr Tyr  
305                 310                 315                 320
Glu Leu Gly Lys Glu Phe Leu Pro Met Glu Ile Gln Leu Ser Arg Leu  
325                 330                 335    
Val Gly Gln Pro Leu Trp Asp Val Ser Arg Ser Ser Thr Gly Asn Leu  
340                 345                 350        
Val Glu Trp Phe Leu Leu Arg Lys Ala Tyr Glu Arg Asn Glu Val Ala  
355                 360                 365            
Pro Asn Lys Pro Ser Glu Glu Glu Tyr Gln Arg Arg Leu Arg Glu Ser  
370                 375                 380                
Tyr Thr Gly Gly Phe Val Lys Glu Pro Glu Lys Gly Leu Trp Glu Asn  
385                 390                 395                 400
Ile Val Tyr Leu Asp Phe Arg Ala Leu Tyr Pro Ser Ile Ile Ile Thr  
405                 410                 415    
His Asn Val Ser Pro Asp Thr Leu Asn Leu Glu Gly Cys Lys Asn Tyr  
420                 425                 430        
Asp Ile Ala Pro Gln Val Gly His Lys Phe Cys Lys Asp Ile Pro Gly  
435                 440                 445            
Phe Ile Pro Ser Leu Leu Gly His Leu Leu Glu Glu Arg Gln Lys Ile  
450                 455                 460                
Lys Thr Lys Met Lys Glu Thr Gln Asp Pro Ile Glu Lys Ile Leu Leu  
465                 470                 475                 480
Asp Tyr Arg Gln Lys Ala Ile Lys Leu Leu Ala Asn Ser Phe Tyr Gly  
485                 490                 495    
Tyr Tyr Gly Tyr Ala Lys Ala Arg Trp Tyr Cys Lys Glu Cys Ala Glu  
500                 505                 510        
Ser Val Thr Ala Trp Gly Arg Lys Tyr Ile Glu Leu Val Trp Lys Glu  
515                 520                 525            
Leu Glu Glu Lys Phe Gly Phe Lys Val Leu Tyr Ile Asp Thr Asp Gly  
530                 535                 540                
Leu Tyr Ala Thr Ile Pro Gly Gly Glu Ser Glu Glu Ile Lys Lys Lys  
545                 550                 555                 560
Ala Leu Glu Phe Val Lys Tyr Ile Asn Ser Lys Leu Pro Gly Leu Leu  
565                 570                 575    
Glu Leu Glu Tyr Glu Gly Phe Tyr Lys Arg Gly Phe Phe Val Thr Lys  
580                 585                 590        
Lys Arg Tyr Ala Val Ile Asp Glu Glu Gly Lys Val Ile Thr Arg Gly  
595                 600                 605            
Leu Glu Ile Val Arg Arg Asp Trp Ser Glu Ile Ala Lys Glu Thr Gln  
610                 615                 620                
Ala Arg Val Leu Glu Thr Ile Leu Lys His Gly Asp Val Glu Glu Ala  
625                 630                 635                 640
Val Arg Ile Val Lys Glu Val Ile Gln Lys Leu Ala Asn Tyr Glu Ile  
645                 650                 655    
Pro Pro Glu Lys Leu Ala Ile Tyr Glu Gln Ile Thr Arg Pro Leu His  
660                 665                 670        
Glu Tyr Lys Ala Ile Gly Pro His Val Ala Val Ala Lys Lys Leu Ala  
675                 680                 685            
Ala Lys Gly Val Lys Ile Lys Pro Gly Met Val Ile Gly Tyr Ile Val  
690                 695                 700                
Leu Arg Gly Asp Gly Pro Ile Ser Asn Arg Ala Ile Leu Ala Glu Glu  
705                 710                 715                 720
Tyr Asp Pro Lys Lys His Lys Tyr Asp Ala Glu Tyr Tyr Ile Glu Asn  
725                 730                 735    
Gln Val Leu Pro Ala Val Leu Arg Ile Leu Glu Gly Phe Gly Tyr Arg  
740                 745                 750        
Lys Glu Asp Leu Arg Tyr Gln Lys Thr Arg Gln Val Gly Leu Thr Ser  
755                 760                 765            
Trp Leu Asn Ile Lys Lys Ser Gly Thr Gly Gly Gly Gly Ala Thr Val  
770                 775                 780                
Lys Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp Ile Ser Lys Ile  
785                 790                 795                 800
Lys Lys Val Trp Arg Val Gly Lys Met Ile Ser Phe Thr Tyr Asp Glu  
805                 810                 815    
Gly Gly Gly Lys Thr Gly Arg Gly Ala Val Ser Glu Lys Asp Ala Pro  
820                 825                 830        
Lys Glu Leu Leu Gln Met Leu Glu Lys Gln Lys Lys                  
835                 840                
<210> 9
<211> 1904
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sac7d-deltaTaq
fusion protein
<400> 9
atgattacga attcgacggt gaaggtaaag ttcaagtata agggtgaaga gaaagaagta 60    
gacacttcaa agataaagaa ggtttggaga gtaggcaaaa tggtgtcctt tacctatgac 120   
gacaatggta agacaggtag aggagctgta agcgagaaag atgctccaaa agaattatta 180   
gacatgttag caagagcaga aagagagaag aaaggcggcg gtgtcactag tcccaaggcc 240   
ctggaggagg ccccctggcc cccgccggaa ggggccttcg tgggctttgt gctttcccgc 300   
aaggagccca tgtgggccga tcttctggcc ctggccgccg ccaggggggg ccgggtccac 360   
cgggcccccg agccttataa agccctcagg gacctgaagg aggcgcgggg gcttctcgcc 420   
aaagacctga gcgttctggc cctgagggaa ggccttggcc tcccgcccgg cgacgacccc 480   
atgctcctcg cctacctcct ggacccttcc aacaccaccc ccgagggggt ggcccggcgc 540   
tacggcgggg agtggacgga ggaggcgggg gagcgggccg ccctttccga gaggctcttc 600   
gccaacctgt gggggaggct tgagggggag gagaggctcc tttggcttta ccgggaggtg 660   
gagaggcccc tttccgctgt cctggcccac atggaggcca cgggggtgcg cctggacgtg 720   
gcctatctca gggccttgtc cctggaggtg gccgaggaga tcgcccgcct cgaggccggg 780   
tcttccgcct ggccggccac cccttcaacc tcaactcccg ggaccagctg gaaagggtcc 840   
tctttgacga gctagggctt cccgccatcg gcaagacgga gaagaccggc aagcgctcca 900   
ccagcgccgc cgtcctggag gccctccgcg aggcccaccc catcgtggag aagatcctgc 960   
agtaccggga gctcaccaag ctgaagagca cctacattga ccccttgccg gacctcatcc 1020  
accccaggac gggccgcctc cacacccgct tcaaccagac ggccacggcc acgggcaggc 1080  
taagtagctc cgatcccaac ctccagaaca tccccgtccg caccccgctt gggcagagga 1140  
tccgccgggc cttcatcgcc gaggaggggt ggctattggt ggccctggac tatagccaga 1200  
tagagctcag ggtgctggcc cacctctccg gcgacgagaa cctgatccgg gtcttccagg 1260  
aggggcggga catccacacg gagaccgcca gctggatgtt cggcgtcccc cgggaggccg 1320  
tggaccccct gatgcgccgg gcggccaaga ccatcaactt cggggtcctc tacggcatgt 1380  
cggcccaccg cctctcccag gagctagcca tcccttacga ggaggcccag gccttcattg 1440  
agcgctactt tcagagcttc cccaaggtgc gggcctggat tgagaagacc ctggaggagg 1500  
gcaggaggcg ggggtacgtg gagaccctct tcggccgccg ccgctacgtg ccagacctag 1560  
aggcccgggt gaagagcgtg cgggaggcgg ccgagcgcat ggccttcaac atgcccgtcc 1620  
agggcaccgc cgccgacctc atgaagctgg ctatggtgaa gctcttcccc aggctggagg 1680  
aaatgggggc caggatgctc cttcaggtcc acgacgagct ggtcctcgag gccccaaaag 1740  
agagggcgga ggccgtggcc cggctggcca aggaggtcat ggagggggtg tatcccctgg 1800  
ccgtgcccct ggaggtggag gtggggatag gggaggactg gctctccgcc aaggagggca 1860  
ttgatggccg cggcggaggc gggcatcatc atcatcatca ttaa                  1904  
<210> 10
<211> 634
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sac7d-deltaTaq
fusion protein
<400> 10
Met Ile Thr Asn Ser Thr Val Lys Val Lys Phe Lys Tyr Lys Gly Glu  
1               5                  10                  15    
Glu Lys Glu Val Asp Thr Ser Lys Ile Lys Lys Val Trp Arg Val Gly  
20                  25                  30        
Lys Met Val Ser Phe Thr Tyr Asp Asp Asn Gly Lys Thr Gly Arg Gly  
35                  40                  45            
Ala Val Ser Glu Lys Asp Ala Pro Lys Glu Leu Leu Asp Met Leu Ala  
50                  55                  60                
Arg Ala Glu Arg Glu Lys Lys Gly Gly Gly Val Thr Ser Pro Lys Ala  
65                  70                  75                  80
Leu Glu Glu Ala Pro Trp Pro Pro Pro Glu Gly Ala Phe Val Gly Phe  
85                  90                  95    
Val Leu Ser Arg Lys Glu Pro Met Trp Ala Asp Leu Leu Ala Leu Ala  
100                 105                 110        
Ala Ala Arg Gly Gly Arg Val His Arg Ala Pro Glu Pro Tyr Lys Ala  
115                 120                 125            
Leu Arg Asp Leu Lys Glu Ala Arg Gly Leu Leu Ala Lys Asp Leu Ser  
130                 135                 140                
Val Leu Ala Leu Arg Glu Gly Leu Gly Leu Pro Pro Gly Asp Asp Pro  
145                 150                 155                 160
Met Leu Leu Ala Tyr Leu Leu Asp Pro Ser Asn Thr Thr Pro Glu Gly  
165                 170                 175    
Val Ala Arg Arg Tyr Gly Gly Glu Trp Thr Glu Glu Ala Gly Glu Arg  
180                 185                 190        
Ala Ala Leu Ser Glu Arg Leu Phe Ala Asn Leu Trp Gly Arg Leu Glu  
195                 200                 205            
Gly Glu Glu Arg Leu Leu Trp Leu Tyr Arg Glu Val Glu Arg Pro Leu  
210                 215                 220                
Ser Ala Val Leu Ala His Met Glu Ala Thr Gly Val Arg Leu Asp Val  
225                 230                 235                 240
Ala Tyr Leu Arg Ala Leu Ser Leu Glu Val Ala Glu Glu Ile Ala Arg  
245                 250                 255    
Leu Glu Ala Glu Val Phe Arg Leu Ala Gly His Pro Phe Asn Leu Asn  
260                 265                 270        
Ser Arg Asp Gln Leu Glu Arg Val Leu Phe Asp Glu Leu Gly Leu Pro  
275                 280                 285            
Ala Ile Gly Lys Thr Glu Lys Thr Gly Lys Arg Ser Thr Ser Ala Ala  
290                 295                 300                
Val Leu Glu Ala Leu Arg Glu Ala His Pro Ile Val Glu Lys Ile Leu  
305                 310                 315                 320
Gln Tyr Arg Glu Leu Thr Lys Leu Lys Ser Thr Tyr Ile Asp Pro Leu  
325                 330                 335    
Pro Asp Leu Ile His Pro Arg Thr Gly Arg Leu His Thr Arg Phe Asn  
340                 345                 350        
Gln Thr Ala Thr Ala Thr Gly Arg Leu Ser Ser Ser Asp Pro Asn Leu  
355                 360                 365            
Gln Asn Ile Pro Val Arg Thr Pro Leu Gly Gln Arg Ile Arg Arg Ala  
370                 375                 380                
Phe Ile Ala Glu Glu Gly Trp Leu Leu Val Ala Leu Asp Tyr Ser Gln  
385                 390                 395                 400
Ile Glu Leu Arg Val Leu Ala His Leu Ser Gly Asp Glu Asn Leu Ile  
405                 410                 415    
Arg Val Phe Gln Glu Gly Arg Asp Ile His Thr Glu Thr Ala Ser Trp  
420                 425                 430        
Met Phe Gly Val Pro Arg Glu Ala Val Asp Pro Leu Met Arg Arg Ala  
435                 440                 445            
Ala Lys Thr Ile Asn Phe Gly Val Leu Tyr Gly Met Ser Ala His Arg  
450                 455                 460                
Leu Ser Gln Glu Leu Ala Ile Pro Tyr Glu Glu Ala Gln Ala Phe Ile  
465                 470                 475                 480
Glu Arg Tyr Phe Gln Ser Phe Pro Lys Val Arg Ala Trp Ile Glu Lys  
485                 490                 495    
Thr Leu Glu Glu Gly Arg Arg Arg Gly Tyr Val Glu Thr Leu Phe Gly  
500                 505                 510        
Arg Arg Arg Tyr Val Pro Asp Leu Glu Ala Arg Val Lys Ser Val Arg  
515                 520                 525            
Glu Ala Ala Glu Arg Met Ala Phe Asn Met Pro Val Gln Gly Thr Ala  
530                 535                 540                
Ala Asp Leu Met Lys Leu Ala Met Val Lys Leu Phe Pro Arg Leu Glu  
545                 550                 555                 560
Glu Met Gly Ala Arg Met Leu Leu Gln Val His Asp Glu Leu Val Leu  
565                 570                 575    
Glu Ala Pro Lys Glu Arg Ala Glu Ala Val Ala Arg Leu Ala Lys Glu  
580                 585                 590        
Val Met Glu Gly Val Tyr Pro Leu Ala Val Pro Leu Glu Val Glu Val  
595                 600                 605            
Gly Ile Gly Glu Asp Trp Leu Ser Ala Lys Glu Gly Ile Asp Gly Arg  
610                 615                 620                
Gly Gly Gly Gly His His His His His His                          
625                 630                
<210> 11
<211> 1965
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:polylysine
(PL)-deltaTaq fusion protein
<400> 11
atgattacga attcgaagaa aaagaaaaag aaaaagcgta agaaacgcaa aaagaaaaag 60    
aaaggcggcg gtgtcactag tggcgcaacc gtaaagttca agtacaaagg cgaagaaaaa 120   
gaggtagaca tctccaagat caagaaagta tggcgtgtgg gcaagatgat ctccttcacc 180   
tacgacgagg gcggtggcaa gaccggccgt ggtgcggtaa gcgaaaagga cgcgccgaag 240   
gagctgctgc agatgctgga gaagcagaaa aagggcggcg gtgtcaccag tcccaaggcc 300   
ctggaggagg ccccctggcc cccgccggaa ggggccttcg tgggctttgt gctttcccgc 360   
aaggagccca tgtgggccga tcttctggcc ctggccgccg ccaggggggg ccgggtccac 420   
cgggcccccg agccttataa agccctcagg gacctgaagg aggcgcgggg gcttctcgcc 480   
aaagacctga gcgttctggc cctgagggaa ggccttggcc tcccgcccgg cgacgacccc 540   
atgctcctcg cctacctcct ggacccttcc aacaccaccc ccgagggggt ggcccggcgc 600   
tacggcgggg agtggacgga ggaggcgggg gagcgggccg ccctttccga gaggctcttc 660   
gccaacctgt gggggaggct tgagggggag gagaggctcc tttggcttta ccgggaggtg 720   
gagaggcccc tttccgctgt cctggcccac atggaggcca cgggggtgcg cctggacgtg 780   
gcctatctca gggccttgtc cctggaggtg gccgaggaga tcgcccgcct cgaggccgag 840   
gtcttccgcc tggccggcca ccccttcaac ctcaactccc gggaccagct ggaaagggtc 900   
ctctttgacg agctagggct tcccgccatc ggcaagacgg agaagaccgg caagcgctcc 960   
accagcgccg ccgtcctgga ggccctccgc gaggcccacc ccatcgtgga gaagatcctg 1020  
cagtaccggg agctcaccaa gctgaagagc acctacattg accccttgcc ggacctcatc 1080  
caccccagga cgggccgcct ccacacccgc ttcaaccaga cggccacggc cacgggcagg 1140  
ctaagtagct ccgatcccaa cctccagaac atccccgtcc gcaccccgct tgggcagagg 1200  
atccgccggg ccttcatcgc cgaggagggg tggctattgg tggccctgga ctatagccag 1260  
atagagctca gggtgctggc ccacctctcc ggcgacgaga acctgatccg ggtcttccag 1320  
gaggggcggg acatccacac ggagaccgcc agctggatgt tcggcgtccc ccgggaggcc 1380  
gtggaccccc tgatgcgccg ggcggccaag accatcaact tcggggtcct ctacggcatg 1440  
tcggcccacc gcctctccca ggagctagcc atcccttacg aggaggccca ggccttcatt 1500  
gagcgctact ttcagagctt ccccaaggtg cgggcctgga ttgagaagac cctggaggag 1560  
ggcaggaggc gggggtacgt ggagaccctc ttcggccgcc gccgctacgt gccagaccta 1620  
gaggcccggg tgaagagcgt gcgggaggcg gccgagcgca tggccttcaa catgcccgtc 1680  
cagggcaccg ccgccgacct catgaagctg gctatggtga agctcttccc caggctggag 1740  
gaaatggggg ccaggatgct ccttcaggtc cacgacgagc tggtcctcga ggccccaaaa 1800  
gagagggcgg aggccgtggc ccggctggcc aaggaggtca tggagggggt gtatcccctg 1860  
gccgtgcccc tggaggtgga ggtggggata ggggaggact ggctctccgc caaggagggc 1920  
attgatggcc gcggcggagg cgggcatcat catcatcatc attaa                 1965  
<210> 12
<211> 654
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:polylysine
(PL)-deltaTaq fusion protein
<400> 12
Met Ile Thr Asn Ser Lys Lys Lys Lys Lys Lys Lys Arg Lys Lys Arg  
1               5                  10                  15    
Lys Lys Lys Lys Lys Gly Gly Gly Val Thr Ser Gly Ala Thr Val Lys  
20                  25                  30        
Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp Ile Ser Lys Ile Lys  
35                  40                  45            
Lys Val Trp Arg Val Gly Lys Met Ile Ser Phe Thr Tyr Asp Glu Gly  
50                  55                  60                
Gly Gly Lys Thr Gly Arg Gly Ala Val Ser Glu Lys Asp Ala Pro Lys  
65                  70                  75                  80
Glu Leu Leu Gln Met Leu Glu Lys Gln Lys Lys Gly Gly Gly Val Thr  
85                  90                  95    
Ser Pro Lys Ala Leu Glu Glu Ala Pro Trp Pro Pro Pro Glu Gly Ala  
100                 105                 110        
Phe Val Gly Phe Val Leu Ser Arg Lys Glu Pro Met Trp Ala Asp Leu  
115                 120                 125            
Leu Ala Leu Ala Ala Ala Arg Gly Gly Arg Val His Arg Ala Pro Glu  
130                 135                 140                
Pro Tyr Lys Ala Leu Arg Asp Leu Lys Glu Ala Arg Gly Leu Leu Ala  
145                 150                 155                 160
Lys Asp Leu Ser Val Leu Ala Leu Arg Glu Gly Leu Gly Leu Pro Pro  
165                 170                 175    
Gly Asp Asp Pro Met Leu Leu Ala Tyr Leu Leu Asp Pro Ser Asn Thr  
180                 185                 190        
Thr Pro Glu Gly Val Ala Arg Arg Tyr Gly Gly Glu Trp Thr Glu Glu  
195                 200                 205            
Ala Gly Glu Arg Ala Ala Leu Ser Glu Arg Leu Phe Ala Asn Leu Trp  
210                 215                 220                
Gly Arg Leu Glu Gly Glu Glu Arg Leu Leu Trp Leu Tyr Arg Glu Val  
225                 230                 235                 240
Glu Arg Pro Leu Ser Ala Val Leu Ala His Met Glu Ala Thr Gly Val  
245                 250                 255    
Arg Leu Asp Val Ala Tyr Leu Arg Ala Leu Ser Leu Glu Val Ala Glu  
260                 265                 270        
Glu Ile Ala Arg Leu Glu Ala Glu Val Phe Arg Leu Ala Gly His Pro  
275                 280                 285            
Phe Asn Leu Asn Ser Arg Asp Gln Leu Glu Arg Val Leu Phe Asp Glu  
290                 295                 300                
Leu Gly Leu Pro Ala Ile Gly Lys Thr Glu Lys Thr Gly Lys Arg Ser  
305                 310                 315                 320
Thr Ser Ala Ala Val Leu Glu Ala Leu Arg Glu Ala His Pro Ile Val  
325                 330                 335    
Glu Lys Ile Leu Gln Tyr Arg Glu Leu Thr Lys Leu Lys Ser Thr Tyr  
340                 345                 350        
Ile Asp Pro Leu Pro Asp Leu Ile His Pro Arg Thr Gly Arg Leu His  
355                 360                 365            
Thr Arg Phe Asn Gln Thr Ala Thr Ala Thr Gly Arg Leu Ser Ser Ser  
370                 375                 380                
Asp Pro Asn Leu Gln Asn Ile Pro Val Arg Thr Pro Leu Gly Gln Arg  
385                 390                 395                 400
Ile Arg Arg Ala Phe Ile Ala Glu Glu Gly Trp Leu Leu Val Ala Leu  
405                 410                 415    
Asp Tyr Ser Gln Ile Glu Leu Arg Val Leu Ala His Leu Ser Gly Asp  
420                 425                 430        
Glu Asn Leu Ile Arg Val Phe Gln Glu Gly Arg Asp Ile His Thr Glu  
435                 440                 445            
Thr Ala Ser Trp Met Phe Gly Val Pro Arg Glu Ala Val Asp Pro Leu  
450                 455                 460                
Met Arg Arg Ala Ala Lys Thr Ile Asn Phe Gly Val Leu Tyr Gly Met  
465                 470                 475                 480
Ser Ala His Arg Leu Ser Gln Glu Leu Ala Ile Pro Tyr Glu Glu Ala  
485                 490                 495    
Gln Ala Phe Ile Glu Arg Tyr Phe Gln Ser Phe Pro Lys Val Arg Ala  
500                 505                 510        
Trp Ile Glu Lys Thr Leu Glu Glu Gly Arg Arg Arg Gly Tyr Val Glu  
515                 520                 525            
Thr Leu Phe Gly Arg Arg Arg Tyr Val Pro Asp Leu Glu Ala Arg Val  
530                 535                 540                
Lys Ser Val Arg Glu Ala Ala Glu Arg Met Ala Phe Asn Met Pro Val  
545                 550                 555                 560
Gln Gly Thr Ala Ala Asp Leu Met Lys Leu Ala Met Val Lys Leu Phe  
565                 570                 575    
Pro Arg Leu Glu Glu Met Gly Ala Arg Met Leu Leu Gln Val His Asp  
580                 585                 590        
Glu Leu Val Leu Glu Ala Pro Lys Glu Arg Ala Glu Ala Val Ala Arg  
595                 600                 605            
Leu Ala Lys Glu Val Met Glu Gly Val Tyr Pro Leu Ala Val Pro Leu  
610                 615                 620                
Glu Val Glu Val Gly Ile Gly Glu Asp Trp Leu Ser Ala Lys Glu Gly  
625                 630                 635                 640
Ile Asp Gly Arg Gly Gly Gly Gly His His His His His His          
645                 650                
<210> 13
<211> 20
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L71F
<400> 13
cctgctctgc cgcttcacgc                                             20    
<210> 14
<211> 20
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L71R
<400> 14
gcacagcggc tggctgagga                                             20    
<210> 15
<211> 22
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L18015F
<400> 15
tgacggagga taacgccagc ag                                          22    
<210> 16
<211> 25
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L23474F
<400> 16
gaaagacgat gggtcgctaa tacgc                                       25    
<210> 17
<211> 22
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L18015F
<400> 17
tgacggagga taacgccagc ag                                          22    
<210> 18
<211> 22
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L29930R
<400> 18
ggggttggag gtcaatgggt tc                                          22    
<210> 19
<211> 20
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L30350F
<400> 19
cctgctctgc cgcttcacgc                                             20    
<210> 20
<211> 22
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L35121R
<400> 20
cacatggtac agcaagcctg gc                                          22    
<210> 21
<211> 23
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L2089F
<400> 21
cccgtatctg ctgggatact ggc                                         23    
<210> 22
<211> 23
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L7112R
<400> 22
cagcggtgct gactgaatca tgg                                         23    
<210> 23
<211> 19
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L30350F
<400> 23
cctgcctgcc gcttcacgc                                              19    
<210> 24
<211> 22
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer L40547R
<400> 24
ccaatacccg tttcatcgcg gc                                          22    
<210> 25
<211> 19
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer
H-Amelo-Y
<400> 25
ccacctcatc ctgggcacc                                              19    
<210> 26
<211> 22
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:primer
H-Amelo-YR
<400> 26
gcttgaggcc aaccatcaga gc                                          22    
<210> 27
<211> 22
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:human
beta-globin primer 536F
<400> 27
ggttggccaa tctactccca gg                                          22    
<210> 28
<211> 20
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:human
beta-globin primer 536R
<400> 28
gctcactcag tgtggcaaag                                             20    
<210> 29
<211> 24
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:human
beta-globin primer 1408R
<400> 29
gattagcaaa agggcctagc ttgg                                        24    
<210> 30
<211> 1899
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial
Sequence:Sso7d-deltaTaq(Y) fusion protein
<400> 30
atgattacga attcgagcgc aaccgtaaag ttcaagtaca aaggcgaaga aaaagaggta 60    
gacatctcca agatcaagaa agtatggcgt gtgggcaaga tgatctcctt cacctacgac 120   
gagggcggtg gcaagaccgg ccgtggtgcg gtaagcgaaa aggacgcgcc gaaggagctg 180   
ctgcagatgc tggagaagca gaaaaagggc ggcggtgtca ctagtcccaa ggccctggag 240   
gaggccccct ggcccccgcc ggaaggggcc ttcgtgggct ttgtgctttc ccgcaaggag 300   
cccatgtggg ccgatcttct ggccctggcc gccgccaggg ggggccgggt ccaccgggcc 360   
cccgagcctt ataaagccct cagggacctg aaggaggcgc gggggcttct cgccaaagac 420   
ctgagcgttc tggccctgag ggaaggcctt ggcctcccgc ccggcgacga ccccatgctc 480   
ctcgcctacc tcctggaccc ttccaacacc acccccgagg gggtggcccg gcgctacggc 540   
ggggagtgga cggaggaggc gggggagcgg gccgcccttt ccgagaggct cttcgccaac 600   
ctgtggggga ggcttgaggg ggaggagagg ctcctttggc tttaccggga ggtggagagg 660   
cccctttccg ctgtcctggc ccacatggag gccacggggg tgcgcctgga cgtggcctat 720   
ctcagggcct tgtccctgga ggtggccgag gagatcgccc gcctcgaggc cgaggtcttc 780   
cgcctggccg gccacccctt caacctcaac tcccgggacc agctggaaag ggtcctcttt 840   
gacgagctag ggcttcccgc catcggcaag acggagaaga ccggcaagcg ctccaccagc 900   
gccgccgtcc tggaggccct ccgcgaggcc caccccatcg tggagaagat cctgcagtac 960   
cgggagctca ccaagctgaa gagcacctac attgacccct tgccggacct catccacccc 1020  
aggacgggcc gcctccacac ccgcttcaac cagacggcca cggccacggg caggctaagt 1080  
agctccgatc ccaacctcca gaacatcccc gtccgcaccc cgcttgggca gaggatccgc 1140  
cgggccttca tcgccgagga ggggtggcta ttggtggccc tggactatag ccagatagag 1200  
ctcagggtgc tggcccacct ctccggcgac gagaacctga tccgggtctt ccaggagggg 1260  
cgggacatcc acacggagac cgccagctgg atgttcggcg tcccccggga ggccgtggac 1320  
cccctgatgc gccgggcggc caagaccatc aactacgggg tcctctacgg catgtcggcc 1380  
caccgcctct cccaggagct agccatccct tacgaggagg cccaggcctt cattgagcgc 1440  
tactttcaga gcttccccaa ggtgcgggcc tggattgaga agaccctgga ggagggcagg 1500  
aggcgggggt acgtggagac cctcttcggc cgccgccgct acgtgccaga cctagaggcc 1560  
cgggtgaaga gcgtgcggga ggcggccgag cgcatggcct tcaacatgcc cgtccagggc 1620  
accgccgccg acctcatgaa gctggctatg gtgaagctct tccccaggct ggaggaaatg 1680  
ggggccagga tgctccttca ggtccacgac gagctggtcc tcgaggcccc aaaagagagg 1740  
gcggaggccg tggcccggct ggccaaggag gtcatggagg gggtgtatcc cctggccgtg 1800  
cccctggagg tggaggtggg gataggggag gactggctct ccgccaagga gggcattgat 1860  
ggccgcggcg gaggcgggca tcatcatcat catcattaa                        1899  
<210> 31
<211> 632
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial
Sequence:Sso7d-deltaTaq(Y) fusion protein
<400> 31
Met Ile Thr Asn Ser Ser Ala Thr Val Lys Phe Lys Tyr Lys Gly Glu  
1               5                  10                  15    
Glu Lys Glu Val Asp Ile Ser Lys Ile Lys Lys Val Trp Arg Val Gly  
20                  25                  30        
Lys Met Ile Ser Phe Thr Tyr Asp Glu Gly Gly Gly Lys Thr Gly Arg  
35                  40                  45            
Gly Ala Val Ser Glu Lys Asp Ala Pro Lys Glu Leu Leu Gln Met Leu  
50                  55                  60                
Glu Lys Gln Lys Lys Gly Gly Gly Val Thr Ser Pro Lys Ala Leu Glu  
65                  70                  75                  80
Glu Ala Pro Trp Pro Pro Pro Glu Gly Ala Phe Val Gly Phe Val Leu  
85                  90                  95    
Ser Arg Lys Glu Pro Met Trp Ala Asp Leu Leu Ala Leu Ala Ala Ala  
100                 105                 110        
Arg Gly Gly Arg Val His Arg Ala Pro Glu Pro Tyr Lys Ala Leu Arg  
115                 120                 125            
Asp Leu Lys Glu Ala Arg Gly Leu Leu Ala Lys Asp Leu Ser Val Leu  
130                 135                 140                
Ala Leu Arg Glu Gly Leu Gly Leu Pro Pro Gly Asp Asp Pro Met Leu  
145                 150                 155                 160
Leu Ala Tyr Leu Leu Asp Pro Ser Asn Thr Thr Pro Glu Gly Val Ala  
165                 170                 175    
Arg Arg Tyr Gly Gly Glu Trp Thr Glu Glu Ala Gly Glu Arg Ala Ala  
180                 185                 190        
Leu Ser Glu Arg Leu Phe Ala Asn Leu Trp Gly Arg Leu Glu Gly Glu  
195                 200                 205            
Glu Arg Leu Leu Trp Leu Tyr Arg Glu Val Glu Arg Pro Leu Ser Ala  
210                 215                 220                
Val Leu Ala His Met Glu Ala Thr Gly Val Arg Leu Asp Val Ala Tyr  
225                 230                 235                 240
Leu Arg Ala Leu Ser Leu Glu Val Ala Glu Glu Ile Ala Arg Leu Glu  
245                 250                 255    
Ala Glu Val Phe Arg Leu Ala Gly His Pro Phe Asn Leu Asn Ser Arg  
260                 265                 270        
Asp Gln Leu Glu Arg Val Leu Phe Asp Glu Leu Gly Leu Pro Ala Ile  
275                 280                 285            
Gly Lys Thr Glu Lys Thr Gly Lys Arg Ser Thr Ser Ala Ala Val Leu  
290                 295                 300                
Glu Ala Leu Arg Glu Ala His Pro Ile Val Glu Lys Ile Leu Gln Tyr  
305                 310                 315                 320
Arg Glu Leu Thr Lys Leu Lys Ser Thr Tyr Ile Asp Pro Leu Pro Asp  
325                 330                 335    
Leu Ile His Pro Arg Thr Gly Arg Leu His Thr Arg Phe Asn Gln Thr  
340                 345                 350        
Ala Thr Ala Thr Gly Arg Leu Ser Ser Ser Asp Pro Asn Leu Gln Asn  
355                 360                 365            
Ile Pro Val Arg Thr Pro Leu Gly Gln Arg Ile Arg Arg Ala Phe Ile  
370                 375                 380                
Ala Glu Glu Gly Trp Leu Leu Val Ala Leu Asp Tyr Ser Gln Ile Glu  
385                 390                 395                 400
Leu Arg Val Leu Ala His Leu Ser Gly Asp Glu Asn Leu Ile Arg Val  
405                 410                 415    
Phe Gln Glu Gly Arg Asp Ile His Thr Glu Thr Ala Ser Trp Met Phe  
420                 425                 430        
Gly Val Pro Arg Glu Ala Val Asp Pro Leu Met Arg Arg Ala Ala Lys  
435                 440                 445            
Thr Ile Asn Tyr Gly Val Leu Tyr Gly Met Ser Ala His Arg Leu Ser  
450                 455                 460                
Gln Glu Leu Ala Ile Pro Tyr Glu Glu Ala Gln Ala Phe Ile Glu Arg  
465                 470                 475                 480
Tyr Phe Gln Ser Phe Pro Lys Val Arg Ala Trp Ile Glu Lys Thr Leu  
485                 490                 495    
Glu Glu Gly Arg Arg Arg Gly Tyr Val Glu Thr Leu Phe Gly Arg Arg  
500                 505                 510        
Arg Tyr Val Pro Asp Leu Glu Ala Arg Val Lys Ser Val Arg Glu Ala  
515                 520                 525            
Ala Glu Arg Met Ala Phe Asn Met Pro Val Gln Gly Thr Ala Ala Asp  
530                 535                 540                
Leu Met Lys Leu Ala Met Val Lys Leu Phe Pro Arg Leu Glu Glu Met  
545                 550                 555                 560
Gly Ala Arg Met Leu Leu Gln Val His Asp Glu Leu Val Leu Glu Ala  
565                 570                 575    
Pro Lys Glu Arg Ala Glu Ala Val Ala Arg Leu Ala Lys Glu Val Met  
580                 585                 590        
Glu Gly Val Tyr Pro Leu Ala Val Pro Leu Glu Val Glu Val Gly Ile  
595                 600                 605            
Gly Glu Asp Trp Leu Ser Ala Lys Glu Gly Ile Asp Gly Arg Gly Gly  
610                 615                 620                
Gly Gly His His His His His His                                  
625                 630        
<210> 32
<211> 2763
<212> DNA
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sso7d-deltaTaq
(E5)(Y) fusion protein
<400> 32
atgattacga attcgagcgc aaccgtaaag ttcaagtaca aaggcgaaga aaaagaggta 60    
gacatctcca agatcaagaa agtatggcgt gtgggcaaga tgatctcctt cacctacgac 120   
gagggcggtg gcaagaccgg ccgtggtgcg gtaagcgaaa aggacgcgcc gaaggagctg 180   
ctgcagatgc tggagaagca gaaaaagggc ggcggtgtca ctagtgggat gctgcccctc 240   
tttgagccca agggccgggt cctcctggtg gacggccacc acctggccta ccgcaccttc 300   
cacgccctga agggcctcac caccagccgg ggggagccgg tgcaggcggt ctacggcttc 360   
gccaagagcc tcctcaaggc cctcaaggag gacggggacg cggtgatcgt ggtctttgac 420   
gccaaggccc cctccttccc ccacgaggcc tacggggggc acaaggcggg ccgggccccc 480   
acgccagagg actttccccg gcaactcgcc ctcatcaagg agctggtgga cctcctgggg 540   
ctggcgcgcc tcgaggtccc gggctacgag gcggacgacg tcctggccag cctggccaag 600   
aaggcggaaa aggagggcta cgaggtccgc atcctcaccg ccgacaaaga cctttaccag 660   
ctcctttccg accgcatcca cgtcctccac cccgaggggt acctcatcac cccggcctgg 720   
ctttgggaaa agtacggcct gaggcccgac cagtgggccg actaccgggc cctgaccggg 780   
gacgagtccg acaaccttcc cggggtcaag ggcatcgggg agaagacggc gaggaagctt 840   
ctggaggagt gggggagcct ggaagccctc ctcaagaacc tggaccggct gaagcccgcc 900   
atccgggaga agatcctggc ccacatggac gatctgaagc tctcctggga cctggccaag 960   
gtgcgcaccg acctgcccct ggaggtggac ttcgccaaaa ggcgggagcc cgaccgggag 1020  
aggcttaggg cctttctgga gaggcttgag tttggcagcc tcctccacga gttcggcctt 1080  
ctggaaagcc ccaaggccct ggaggaggcc ccctggcccc cgccggaagg ggccttcgtg 1140  
ggctttgtgc tttcccgcaa ggagcccatg tgggccgatc ttctggccct ggccgccgcc 1200  
agggggggcc gggtccaccg ggcccccgag ccttataaag ccctcaggga cctgaaggag 1260  
gcgcgggggc ttctcgccaa agacctgagc gttctggccc tgagggaagg ccttggcctc 1320  
ccgcccggcg acgaccccat gctcctcgcc tacctcctgg acccttccaa caccaccccc 1380  
gagggggtgg cccggcgcta cggcggggag tggacggagg aggcggggga gcgggccgcc 1440  
ctttccgaga ggctcttcgc caacctgtgg gggaggcttg agggggagga gaggctcctt 1500  
tggctttacc gggaggtgga gaggcccctt tccgctgtcc tggcccacat ggaggccacg 1560  
ggggtgcgcc tggacgtggc ctatctcagg gccttgtccc tggaggtggc cgaggagatc 1620  
gcccgcctcg aggccgaggt cttccgcctg gccggccacc ccttcaacct caactcccgg 1680  
gaccagctgg aaagggtcct ctttgacgag ctagggcttc ccgccatcgg caagacggag 1740  
aagaccggca agcgctccac cagcgccgcc gtcctggagg ccctccgcga ggcccacccc 1800  
atcgtggaga agatcctgca gtaccgggag ctcaccaagc tgaagagcac ctacattgac 1860  
cccttgccgg acctcatcca ccccaggacg ggccgcctcc acacccgctt caaccagacg 1920  
gccacggcca cgggcaggct aagtagctcc gatcccaacc tccagaacat ccccgtccgc 1980  
accccgcttg ggcagaggat ccgccgggcc ttcatcgccg aggaggggtg gctattggtg 2040  
gccctggact atagccagat agagctcagg gtgctggccc acctctccgg cgacgagaac 2100  
ctgatccggg tcttccagga ggggcgggac atccacacgg agaccgccag ctggatgttc 2160  
ggcgtccccc gggaggccgt ggaccccctg atgcgccggg cggccaagac catcaactac 2220  
ggggtcctct acggcatgtc ggcccaccgc ctctcccagg agctagccat cccttacgag 2280  
gaggcccagg ccttcattga gcgctacttt cagagcttcc ccaaggtgcg ggcctggatt 2340  
gagaagaccc tggaggaggg caggaggcgg gggtacgtgg agaccctctt cggccgccgc 2400  
cgctacgtgc cagacctaga ggcccgggtg aagagcgtgc gggaggcggc cgagcgcatg 2460  
gccttcaaca tgcccgtcca gggcaccgcc gccgacctca tgaagctggc tatggtgaag 2520  
ctcttcccca ggctggagga aatgggggcc aggatgctcc ttcaggtcca cgacgagctg 2580  
gtcctcgagg ccccaaaaga gagggcggag gccgtggccc ggctggccaa ggaggtcatg 2640  
gagggggtgt atcccctggc cgtgcccctg gaggtggagg tggggatagg ggaggactgg 2700  
ctctccgcca aggagggcat tgatggccgc ggcggaggcg ggcatcatca tcatcatcat 2760  
taa                                                               2763  
<210> 33
<211> 920
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:Sso7d-deltaTaq
(E5)(Y) fusion protein
<400> 33
Met Ile Thr Asn Ser Ser Ala Thr Val Lys Phe Lys Tyr Lys Gly Glu  
1               5                  10                  15    
Glu Lys Glu Val Asp Ile Ser Lys Ile Lys Lys Val Trp Arg Val Gly  
20                  25                  30        
Lys Met Ile Ser Phe Thr Tyr Asp Glu Gly Gly Gly Lys Thr Gly Arg  
35                  40                  45            
Gly Ala Val Ser Glu Lys Asp Ala Pro Lys Glu Leu Leu Gln Met Leu  
50                  55                  60                
Glu Lys Gln Lys Lys Gly Gly Gly Val Thr Ser Gly Met Leu Pro Leu  
65                  70                  75                  80
Phe Glu Pro Lys Gly Arg Val Leu Leu Val Asp Gly His His Leu Ala  
85                  90                  95    
Tyr Arg Thr Phe His Ala Leu Lys Gly Leu Thr Thr Ser Arg Gly Glu  
100                 105                 110        
Pro Val Gln Ala Val Tyr Gly Phe Ala Lys Ser Leu Leu Lys Ala Leu  
115                 120                 125            
Lys Glu Asp Gly Asp Ala Val Ile Val Val Phe Asp Ala Lys Ala Pro  
130                 135                 140                
Ser Phe Pro His Glu Ala Tyr Gly Gly His Lys Ala Gly Arg Ala Pro  
145                 150                 155                 160
Thr Pro Glu Asp Phe Pro Arg Gln Leu Ala Leu Ile Lys Glu Leu Val  
165                 170                 175    
Asp Leu Leu Gly Leu Ala Arg Leu Glu Val Pro Gly Tyr Glu Ala Asp  
180                 185                 190        
Asp Val Leu Ala Ser Leu Ala Lys Lys Ala Glu Lys Glu Gly Tyr Glu  
195                 200                 205            
Val Arg Ile Leu Thr Ala Asp Lys Asp Leu Tyr Gln Leu Leu Ser Asp  
210                 215                 220                
Arg Ile His Val Leu His Pro Glu Gly Tyr Leu Ile Thr Pro Ala Trp  
225                 230                 235                 240
Leu Trp Glu Lys Tyr Gly Leu Arg Pro Asp Gln Trp Ala Asp Tyr Arg  
245                 250                 255    
Ala Leu Thr Gly Asp Glu Ser Asp Asn Leu Pro Gly Val Lys Gly Ile  
260                 265                 270        
Gly Glu Lys Thr Ala Arg Lys Leu Leu Glu Glu Trp Gly Ser Leu Glu  
275                 280                 285            
Ala Leu Leu Lys Asn Leu Asp Arg Leu Lys Pro Ala Ile Arg Glu Lys  
290                 295                 300                
Ile Leu Ala His Met Asp Asp Leu Lys Leu Ser Trp Asp Leu Ala Lys  
305                 310                 315                 320
Val Arg Thr Asp Leu Pro Leu Glu Val Asp Phe Ala Lys Arg Arg Glu  
325                 330                 335    
Pro Asp Arg Glu Arg Leu Arg Ala Phe Leu Glu Arg Leu Glu Phe Gly  
340                 345                 350        
Ser Leu Leu His Glu Phe Gly Leu Leu Glu Ser Pro Lys Ala Leu Glu  
355                 360                 365            
Glu Ala Pro Trp Pro Pro Pro Glu Gly Ala Phe Val Gly Phe Val Leu  
370                 375                 380                
Ser Arg Lys Glu Pro Met Trp Ala Asp Leu Leu Ala Leu Ala Ala Ala  
385                 390                 395                 400
Arg Gly Gly Arg Val His Arg Ala Pro Glu Pro Tyr Lys Ala Leu Arg  
405                 410                 415    
Asp Leu Lys Glu Ala Arg Gly Leu Leu Ala Lys Asp Leu Ser Val Leu  
420                 425                 430        
Ala Leu Arg Glu Gly Leu Gly Leu Pro Pro Gly Asp Asp Pro Met Leu  
435                 440                 445            
Leu Ala Tyr Leu Leu Asp Pro Ser Asn Thr Thr Pro Glu Gly Val Ala  
450                 455                 460                
Arg Arg Tyr Gly Gly Glu Trp Thr Glu Glu Ala Gly Glu Arg Ala Ala  
465                 470                 475                 480
Leu Ser Glu Arg Leu Phe Ala Asn Leu Trp Gly Arg Leu Glu Gly Glu  
485                 490                 495    
Glu Arg Leu Leu Trp Leu Tyr Arg Glu Val Glu Arg Pro Leu Ser Ala  
500                 505                 510        
Val Leu Ala His Met Glu Ala Thr Gly Val Arg Leu Asp Val Ala Tyr  
515                 520                 525            
Leu Arg Ala Leu Ser Leu Glu Val Ala Glu Glu Ile Ala Arg Leu Glu  
530                 535                 540                
Ala Glu Val Phe Arg Leu Ala Gly His Pro Phe Asn Leu Asn Ser Arg  
545                 550                 555                 560
Asp Gln Leu Glu Arg Val Leu Phe Asp Glu Leu Gly Leu Pro Ala Ile  
565                 570                 575    
Gly Lys Thr Glu Lys Thr Gly Lys Arg Ser Thr Ser Ala Ala Val Leu  
580                 585                 590        
Glu Ala Leu Arg Glu Ala His Pro Ile Val Glu Lys Ile Leu Gln Tyr  
595                 600                 605            
Arg Glu Leu Thr Lys Leu Lys Ser Thr Tyr Ile Asp Pro Leu Pro Asp  
610                 615                 620                
Leu Ile His Pro Arg Thr Gly Arg Leu His Thr Arg Phe Asn Gln Thr  
625                 630                 635                 640
Ala Thr Ala Thr Gly Arg Leu Ser Ser Ser Asp Pro Asn Leu Gln Asn  
645                 650                 655    
Ile Pro Val Arg Thr Pro Leu Gly Gln Arg Ile Arg Arg Ala Phe Ile  
660                 665                 670        
Ala Glu Glu Gly Trp Leu Leu Val Ala Leu Asp Tyr Ser Gln Ile Glu  
675                 680                 685            
Leu Arg Val Leu Ala His Leu Ser Gly Asp Glu Asn Leu Ile Arg Val  
690                 695                 700                
Phe Gln Glu Gly Arg Asp Ile His Thr Glu Thr Ala Ser Trp Met Phe  
705                 710                 715                 720
Gly Val Pro Arg Glu Ala Val Asp Pro Leu Met Arg Arg Ala Ala Lys  
725                 730                 735    
Thr Ile Asn Tyr Gly Val Leu Tyr Gly Met Ser Ala His Arg Leu Ser  
740                 745                 750        
Gln Glu Leu Ala Ile Pro Tyr Glu Glu Ala Gln Ala Phe Ile Glu Arg  
755                 760                 765            
Tyr Phe Gln Ser Phe Pro Lys Val Arg Ala Trp Ile Glu Lys Thr Leu  
770                 775                 780                
Glu Glu Gly Arg Arg Arg Gly Tyr Val Glu Thr Leu Phe Gly Arg Arg  
785                 790                 795                 800
Arg Tyr Val Pro Asp Leu Glu Ala Arg Val Lys Ser Val Arg Glu Ala  
805                 810                 815    
Ala Glu Arg Met Ala Phe Asn Met Pro Val Gln Gly Thr Ala Ala Asp  
820                 825                 830        
Leu Met Lys Leu Ala Met Val Lys Leu Phe Pro Arg Leu Glu Glu Met  
835                 840                 845            
Gly Ala Arg Met Leu Leu Gln Val His Asp Glu Leu Val Leu Glu Ala  
850                 855                 860                
Pro Lys Glu Arg Ala Glu Ala Val Ala Arg Leu Ala Lys Glu Val Met  
865                 870                 875                 880
Glu Gly Val Tyr Pro Leu Ala Val Pro Leu Glu Val Glu Val Gly Ile  
885                 890                 895    
Gly Glu Asp Trp Leu Ser Ala Lys Glu Gly Ile Asp Gly Arg Gly Gly  
900                 905                 910        
Gly Gly His His His His His His                                  
915                 920
<210> 34
<211> 4
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:peptide linker
<400> 34
Gly Gly Val Thr                                                  
1            
<210> 35
<211> 6
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:peptide linker
<400> 35
Gly Thr Gly Gly Gly Gly                                          
1               5    
<210> 36
<211> 21
<212> PRT
<213> Artificial Sequence
<220> 
<223> Description of Artificial Sequence:lysine-rich
peptide
<400> 36
Asn Ser Lys Lys Lys Lys Lys Lys Lys Arg Lys Lys Arg Lys Lys Lys  
1               5                  10                  15    
Gly Gly Gly Val Thr                                              
20