KR100847386B1 - 소마토트로핀의 발현을 위한 재조합 ｄｎａ 벡터 - Google Patents
소마토트로핀의 발현을 위한 재조합 ｄｎａ 벡터 Download PDFInfo
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Abstract
미생물, 바람직하게는, 제한되지는 않으나 대장균(E.coli)과 같은 박테리아를 형질전환시켜, 통상적인 배양 및 유도 조건을 사용하여 높은 수준의 소 소마토트로핀 발현이 가능하도록 하는 일련의 소마토트로핀 벡터족이 제공된다.
소마토트로핀, 형질전환, 벡터
Description
본 발명은, 소마토트로핀의 발현을 위한 재조합 DNA 벡터에 관한 것이다.
관련 출원에 대한 상호참증
본 출원은 35 U.S.C.119(e)에 기초하여, 2000년 12월 26일에 출원된 출원번호 60/258,291에 대한 우선권을 주장하며, 상기 우선권 기초출원은 전체가 여기에 참조문헌으로 포함되었다.
포유류에서, 성장호르몬(소마토트로핀)은 처음에 리더 서열을 포함하는 전호르몬(pre-hormone) 형태로 발현되며, 이 리더 서열은 뇌하수체 세포에서 성숙하고, 생물학적으로 활성인 호르몬으로 분비되는 동안 제거된다. 본 명세서에서 사용되는 "성숙한" 소마토트로핀("ST")이란 동물의 혈액으로 분비되는 ST의 아미노산 길이와 반드시 같은 길이를 갖는 것을 의미하며, 예로서, N-Ala-Phe-Pro- 또는 N-Phe-Pro-소 ST, N-Phe-Pro-인간, -돼지(개와 동일) 또는 -말 ST 등이 있다.
인간 및 소의 ST를 cDNA 서열을 가지는 구조유전자를 사용하여 비분비 시스템으로 박테리아에서 발현시키기 위한 시도는 처음에는 실패하였다. 연구자들은 상기 구조유전자의 앞쪽 말단으로 침묵 돌연변이(silent mutation)를 도입시킨 후에 성공하였다. P.H. Seeburg 등은 초기의 어려움은 cDNA에 상응하는 mRNA의 2차 구조에 의해 발현이 저해되었기 때문이라고 생각하였고, 상기 2차 구조를 느슨하게 하여 중요한 발현을 가능케한 방법을 가르쳐주었다. DNA, Vol. 2, No. 1, 1983, pp.37-45; 미국특허 제5,254,463호 및 제5,260,201호.
어떠한 침묵 돌연변이(silent mutation)를 포함하는 DNA를 사용하여 상업적으로 실용가능한 ST 발현 수준을 얻을 수 있게 되었다. 그러나, 실용을 위한 생산성이 매우 높고, 저렴한 새로운 벡터에 대한 요구를 만족시키기 어려웠으며, 이는 성숙한 소마토트로핀 벡터의 구조와 발현 수준사이의 관계가 잘 알려지지 않았고, 예측이 매우 어려웠기 때문이다.
발명의 요약
본 명세서에 설명된 것은 일련의 ST 벡터족으로서, 통상적인 배양 및 유도 조건을 사용하여 높은 수준으로 소 ST("bST")을 발현시키기 위해 상기 벡터로 미생물, 바람직하게는 제한되지는 않으나 대장균(E.coli)과 같은 박테리아가 형질전환되어 질 수 있다. 상기 벡터족은 서열목록에 서열 번호 1~44인 44종의 벡터로 예시되어있다. 각 염기서열은 프로모터 영역의 앞쪽에 위치한 EcoRⅠ부위(GAATTC)부터 프로모터, 리보좀결합부위("RBS") 및 전체 bST 구조 유전자를 거쳐 상기 구조 유전자에 대한 번역 종결코돈(stop codon)으로 끝난다.
발명의 상세한 설명
서열 번호 1~44를 가지는 각 벡터는 합성 프로모터("cpex-20")를 포함하며, 상기 프로모터는 1999년 4월 1일에 출원된 미국 가특허출원 60/127,449호에 대한 우선권을 주장하여, 2000년 3월 31일 PCT출원된 PCT/US00/40014에 공개되어 있다. 상기 프로모터는 EcoRⅠ부위(GAATTC)와 AscⅠ부위(GGCGCGCC) 사이에 위치한다. 다양한 다른 통상적인 프로모터 및 새로운 프로모터가 우수한 ST 발현 수준을 위해 cpex-20 대신에 이들 벡터에 사용될 수 있다.
서열 번호 1~17은 대장균(E.coli) 유래의 동일한 RBS(이하,"dps")를 가진다. 서열 번호 18~29는 새로운 합성 RBS(이하,"L49")를 공유한다. 서열 번호 30~42는 제2의 새로운 합성 RBS(이하,"L437")를 공유한다. 서열 번호 43~44는 제3의 새로운 합성 RBS(이하,"R806")를 공유한다. 각 RBS는 AscⅠ부위와 bST 구조 유전자의 ATG 번역 개시코돈 사이에 위치한다. 상기 AscⅠ부위는 각 서열의 동위의 76번째 염기에서 시작된다. 상기 ATG 번역 개시코돈은 대략 동위의 109번째 염기에서 시작되며; 이는 상기 RBS가 약간씩 다른 길이를 가지므로 다소 차이가 날 수 있다.
상기 dps, L49, L437 및 L806의 구체적인 염기서열은 다음과 같다:
dps: cataacatca agaggatatg aaatt(서열 번호 1~17의 84번째 염기부터 108번째 염기까지)
L49: gtacaaatca tagagggtat ttaat(서열 번호 18~29의 84번째 염기부터 108번째 염기까지)
L437: agacactaaa tagagggtat ttaatt(서열 번호 30~42의 84번째 염기부터 109번째 염기까지)
L806: taaaagagac taggaggaga ttaga(서열 번호 43~44의 84번째 염기부터 108번째 염기까지)
상기 dps, L49, L437 및 L806의 구체적인 염기서열은 다음과 같다:
dps: cataacatca agaggatatg aaatt(서열 번호 1~17의 84번째 염기부터 108번째 염기까지)
L49: gtacaaatca tagagggtat ttaat(서열 번호 18~29의 84번째 염기부터 108번째 염기까지)
L437: agacactaaa tagagggtat ttaatt(서열 번호 30~42의 84번째 염기부터 109번째 염기까지)
L806: taaaagagac taggaggaga ttaga(서열 번호 43~44의 84번째 염기부터 108번째 염기까지)
서열 번호 1~44를 가지는 모든 벡터들은 bST의 처음 16개의 코돈을 인코딩하는 뉴클레오타이드 내에 다양한 일련의 침묵 돌연변이(silent mutation)를 포함하므로써, "성숙한" bST를 인코딩하는 cDNA와 다른 구조 유전자를 가진다. 도 1에 나타냈듯이, 벡터 pXT709(서열 번호 16)는 cDNA의 구조 유전자로부터 변형된 구조 유전자 내에, 원하지 않는 제한효소 인지부위를 제거하기 위해 대장균(E.coli)이 선호하는 11개의 코돈 및 12번째 코돈으로 된 12개의 코돈을 추가로 가지며, 1개의 번역 종결코돈을 2개의 나란한 종결코돈으로 바꾸고, 일렬로 나란한 2개의 lacUV5 프로모터를 가지는 다른 종결자(terminator)로 대체하므로써 pXT674(서열 번호 8)와는 다르다. pXT703(서열 번호 15) 및 pXT747(서열 번호 17)은 각각 상기와 같은 면에서 pXT601(서열 번호 2) 및 pXT686(서열 번호 13)과 다르다.
서열 번호 8과 서열 번호 16에서 bST의 처음 16개의 코돈을 인코딩하는 뉴클레오티드 내의 침묵 돌연변이의 구체적인 위치 및 변형 염기를 야생형(wild type)과 비교하면 다음과 같다:
서열 번호 8과 서열 번호 16에서 bST의 처음 16개의 코돈을 인코딩하는 뉴클레오티드 내의 침묵 돌연변이의 구체적인 위치 및 변형 염기를 야생형(wild type)과 비교하면 다음과 같다:
서열 번호 1~44를 가지는 각 벡터는 유도체로서 인돌아크릴산(indole acrylic acid) 대신 50ppm의 나리딕신산(nalidixic acid)이 사용되는 외에는 Bogosian 등, J.Biol.Chem., vol.264, pp.531-39, January 5,1989에 설명된 배양 조건을 사용하여, 최적화가 아닌 상태에서는 성숙한 bST를 적어도 약 3.5gm/l, 이들 대부분은 4.5gm/l이상, 일부는 7gm/l 또는 그이상 발현한다.
성숙한 ST 벡터들의 발현 수준은 간행된 방법, 예컨데, P.H. Seeburg 등으로부터 예측이 매우 어렵다. 사실, 서열 번호 1~17, 30~35, 및 37~42인 벡터들은 Seeburg 등에서 권유한 계산법을 사용하면, bST cDNA의 mRNA로부터 느슨해 졌다기 보다는, 더 커진 mRNA 2차구조를 가지는 것으로 밝혀졌다.
도 1은 벡터 pXT674(서열 번호 8) 및 pXT709(서열 번호 16)의 DNA 염기서열을 나타낸 도면이다.
<110> MONSANTO TECHNOLOGY LLC
Bogosian, Gregg
O'Neil, Julia P.
Aardema, Noelle D.
<120> RECOMBINANT DNA VECTORS FOR EXPRESSION OF SOMATOTROPINS
<130> 37-21(52296)B
<150> US 60/258,291
<151> 2000-12-26
<160> 44
<170> PatentIn version 3.1
<210> 1
<211> 684
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<213> Artificial Sequence
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<400> 1
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagca 120
atgagcttgt ccggactttt tgcaaatgct gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
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gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
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atgtcattgt ccggactgtt tgcgaacgct gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
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gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttccctgca 120
atgtcattgt ccggactgtt tgcaaatgcg gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
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gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
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gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
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atgagcttgt ccggattgtt tgcaaacgct gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
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gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc 120
atgtcattgt ccggactgtt tgcaaatgct gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 7
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 673
<400> 7
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc 120
atgagcttgt ccggactgtt tgccaacgct gtcctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 8
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 674
<400> 8
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc 120
atgagcttgt ccggactctt tgccaatgct gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 9
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 675
<400> 9
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc 120
atgagcttgt ccggactgtt tgccaatgct gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 10
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 676
<400> 10
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc 120
atgtccttgt ccggactctt tgccaacgct gtcctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 11
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 682
<400> 11
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc 120
atgtcattgt ccggactgtt tgcgaacgct gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 12
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 683
<400> 12
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc 120
atgtcattgt ccggactctt tgcaaatgct gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 13
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 686
<400> 13
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc 120
atgtcattgt ccggactctt tgcgaacgct gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 14
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 689
<400> 14
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc 120
atgtcattgt ccggactgtt tgcaaacgct gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 15
<211> 687
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 703
<400> 15
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc 120
atgtcattgt ccggactgtt tgcgaacgct gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gcgttactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc gacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
agctggcttg ggccgctgca gttcctcagc cgtgtcttca ccaacagctt ggtgtttggc 420
accagcgacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgtgagctgg aagatggcac cccgcgtgct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccgtaagg acctgcataa gacggagacg tacctgcgtg tcatgaagtg ccgccgcttc 660
ggggaggcga gctgcgcctt ctaatag 687
<210> 16
<211> 687
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 709
<400> 16
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gtttccagcc 120
atgagcttgt ccggactctt tgccaatgct gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gcgttactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc gacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
agctggcttg ggccgctgca gttcctcagc cgtgtcttca ccaacagctt ggtgtttggc 420
accagcgacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgtgagctgg aagatggcac cccgcgtgct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccgtaagg acctgcataa gacggagacg tacctgcgtg tcatgaagtg ccgccgcttc 660
ggggaggcga gctgcgcctt ctaatag 687
<210> 17
<211> 687
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 747
<400> 17
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcccataaca tcaagaggat atgaaattat gttcccagcc 120
atgtcattgt ccggactctt tgcgaacgct gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gcgttactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc gacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
agctggcttg ggccgctgca gttcctcagc cgtgtcttca ccaacagctt ggtgtttggc 420
accagcgacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgtgagctgg aagatggcac cccgcgtgct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccgtaagg acctgcataa gacggagacg tacctgcgtg tcatgaagtg ccgccgcttc 660
ggggaggcga gctgcgcctt ctaatag 687
<210> 18
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 487
<400> 18
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttcctgcg 120
atgtcattgt ccggattgtt tgcgaatgcg gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 19
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 536
<400> 19
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca 120
atgtcattgt ccggattgtt tgcaaatgcc gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 20
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 541
<400> 20
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca 120
atgtcattgt ccggattgtt tgcgaacgcc gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 21
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 542
<400> 21
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttcctgca 120
atgtcattgt ccggactttt tgccaacgca gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 22
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 548
<400> 22
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca 120
atgtcattgt ccggactctt tgcaaacgcc gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 23
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 549
<400> 23
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttcctgca 120
atgtccttgt ccggactttt tgccaatgca gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 24
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 562
<400> 24
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagcc 120
atgtcattgt ccggactttt tgccaacgca gtcctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 25
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 576
<400> 25
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca 120
atgtcattgt ccggactgtt tgcgaacgcg gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 26
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 579
<400> 26
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca 120
atgtcattgt ccggactttt tgccaacgcg gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 27
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 580
<400> 27
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca 120
atgtcattgt ccggactgtt tgccaacgcc gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 28
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 581
<400> 28
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagcc 120
atgtccttgt ccggactttt tgccaacgca gtcctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 29
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 584
<400> 29
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccgtacaaa tcatagaggg tatttaatat gtttccagca 120
atgtcattgt ccggactttt tgcgaatgct gtactccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 30
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 491
<400> 30
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc 120
catgtccttg tccggattgt ttgcgaatgc ggtcctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 31
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 529
<400> 31
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc 120
aatgagcttg tccggattgt ttgcgaatgc ggtgctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 32
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 530
<400> 32
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttccagc 120
catgtcattg tccggactct ttgcgaatgc ggtcctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 33
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 619
<400> 33
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc 120
gatgagcttg tccggattgt ttgcgaatgc agtgctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 34
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 605
<400> 34
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttccagc 120
aatgtccttg tccggactgt ttgcgaacgc agtcctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 35
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 606
<400> 35
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc 120
aatgtccttg tccggattgt ttgcgaacgc ggtcctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 36
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 607
<400> 36
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc 120
catgtccttg tccggactct ttgccaacgc tgtgctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 37
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 620
<400> 37
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc 120
aatgagcttg tccggacttt ttgcgaatgc tgtactccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 38
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 631
<400> 38
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc 120
catgagcttg tccggattgt ttgcaaacgc cgtcctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 39
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 632
<400> 39
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttccagc 120
catgtcattg tccggattgt ttgcaaacgc tgtcctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 40
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 633
<400> 40
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttccagc 120
catgtcattg tccggacttt ttgcgaacgc tgtgctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 41
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 635
<400> 41
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgtttcctgc 120
catgtcattg tccggacttt ttgccaatgc ggtcctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 42
<211> 685
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 681
<400> 42
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gccagacact aaatagaggg tatttaatta tgttcccagc 120
catgtcattg tccggacttt ttgccaacgc ggtgctccgg gctcagcacc tgcatcagct 180
ggctgctgac accttcaaag agtttgagcg cacctacatc ccggagggac agagatactc 240
catccagaac acccaggttg ccttctgctt ctctgaaacc atcccggccc ccacgggcaa 300
gaatgaggcc cagcagaaat cagacttgga gctgcttcgc atctcactgc tcctcatcca 360
gtcgtggctt gggcccctgc agttcctcag cagagtcttc accaacagct tggtgtttgg 420
cacctcggac cgtgtctatg agaagctgaa ggacctggag gaaggcatcc tggccctgat 480
gcgggagctg gaagatggca ccccccgggc tgggcagatc ctcaagcaga cctatgacaa 540
atttgacaca aacatgcgca gtgacgacgc gctgctcaag aactacggtc tgctctcctg 600
cttccggaag gacctgcata agacggagac gtacctgagg gtcatgaagt gccgccgctt 660
cggggaggcc agctgcgcct tctag 685
<210> 43
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 483
<400> 43
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcctaaaaga gactaggagg agattagaat gttcccagca 120
atgagtttgt ccggactgtt tgcgaacgcc gtgctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
<210> 44
<211> 684
<212> DNA
<213> Artificial Sequence
<220>
<223> Somatotropin expression vector pXT 564
<400> 44
gaattctact gtatgagcat acagttaagg gttgacaacc gatatttatt cacttaatat 60
ataaatatca actgaggcgc gcctaaaaga gactaggagg agattagaat gttcccagca 120
atgtcattgt ccggactgtt tgccaatgcg gtcctccggg ctcagcacct gcatcagctg 180
gctgctgaca ccttcaaaga gtttgagcgc acctacatcc cggagggaca gagatactcc 240
atccagaaca cccaggttgc cttctgcttc tctgaaacca tcccggcccc cacgggcaag 300
aatgaggccc agcagaaatc agacttggag ctgcttcgca tctcactgct cctcatccag 360
tcgtggcttg ggcccctgca gttcctcagc agagtcttca ccaacagctt ggtgtttggc 420
acctcggacc gtgtctatga gaagctgaag gacctggagg aaggcatcct ggccctgatg 480
cgggagctgg aagatggcac cccccgggct gggcagatcc tcaagcagac ctatgacaaa 540
tttgacacaa acatgcgcag tgacgacgcg ctgctcaaga actacggtct gctctcctgc 600
ttccggaagg acctgcataa gacggagacg tacctgaggg tcatgaagtg ccgccgcttc 660
ggggaggcca gctgcgcctt ctag 684
Claims (6)
- 대장균(E.coli)에서의 성숙한 소 소마토트로핀의 발현에 유용한, 서열 번호 8 또는 서열 번호 16을 포함하는 벡터.
- 제 1항에 있어서, 서열 번호 8을 포함하는 벡터.
- 제 1항에 있어서, 서열 번호 16을 포함하는 벡터.
- 제 1항 내지 제3항 중 어느 한 항의 벡터로 형질전환된 대장균.
- 제 4항에 따른 대장균의 배양 및 소 소마토트로핀의 발현 유도를 포함하는 성숙한 소 소마토트로핀의 제조방법.
- 삭제
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US25829100P | 2000-12-26 | 2000-12-26 | |
| US60/258,291 | 2000-12-26 | ||
| PCT/US2001/049977 WO2002051238A2 (en) | 2000-12-26 | 2001-12-21 | Recombinant dna vectors for expression of somatotropins |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20030074679A KR20030074679A (ko) | 2003-09-19 |
| KR100847386B1 true KR100847386B1 (ko) | 2008-07-18 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020037008578A KR100847386B1 (ko) | 2000-12-26 | 2001-12-21 | 소마토트로핀의 발현을 위한 재조합 ｄｎａ 벡터 |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US6828124B2 (ko) |
| KR (1) | KR100847386B1 (ko) |
| AT (1) | AT500850B1 (ko) |
| AU (1) | AU2002232758A1 (ko) |
| WO (1) | WO2002051238A2 (ko) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1702074B1 (en) | 2003-09-25 | 2009-11-11 | Monsanto Technology LLC | Prevention of incorporation of non-standard amino acids into protein |
| CA2622710C (en) | 2005-09-16 | 2016-10-18 | Monsanto Technology Llc | Hybrid portable origin of replication plasmids |
| WO2007142954A2 (en) * | 2006-05-30 | 2007-12-13 | Dow Global Technologies Inc. | Codon optimization method |
| EP2363495B1 (en) * | 2006-05-30 | 2019-07-24 | Pfenex Inc. | Anthrax vaccine |
| US20110129873A1 (en) * | 2008-04-30 | 2011-06-02 | Monsanto Technology Llc | Recombinant DNA Vectors for Expression of Human Prolactin Antagonists |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0534705A2 (en) | 1991-09-24 | 1993-03-31 | Eli Lilly And Company | Expression of low molecular weight recombinant polypeptides |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IE52755B1 (en) | 1980-08-26 | 1988-02-17 | Univ California | Bovine pre-growth and growth hormone |
| US5254463A (en) | 1981-09-18 | 1993-10-19 | Genentech, Inc. | Method for expression of bovine growth hormone |
| IL104086A (en) * | 1991-12-18 | 1998-03-10 | Lilly Co Eli | Translational activating oligonucleotide sequence useful for the high level expression of polypeptides and a method for producing such polypeptides |
| WO2000060103A2 (en) * | 1999-04-01 | 2000-10-12 | Monsanto Technology Llc | Dna construct comprising a cyanophage of cyanobacteria promoter and its use |
-
2001
- 2001-12-21 KR KR1020037008578A patent/KR100847386B1/ko active IP Right Grant
- 2001-12-21 AT AT0928701A patent/AT500850B1/de not_active IP Right Cessation
- 2001-12-21 WO PCT/US2001/049977 patent/WO2002051238A2/en not_active Application Discontinuation
- 2001-12-21 AU AU2002232758A patent/AU2002232758A1/en not_active Abandoned
-
2002
- 2002-06-17 US US10/173,082 patent/US6828124B2/en not_active Expired - Lifetime
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0534705A2 (en) | 1991-09-24 | 1993-03-31 | Eli Lilly And Company | Expression of low molecular weight recombinant polypeptides |
Non-Patent Citations (1)
| Title |
|---|
| Biotechnology (NY) vol.9(9):869-872 (1991.09.)* |
Also Published As
| Publication number | Publication date |
|---|---|
| AT500850A1 (de) | 2006-04-15 |
| WO2002051238A3 (en) | 2003-08-21 |
| KR20030074679A (ko) | 2003-09-19 |
| AU2002232758A1 (en) | 2002-07-08 |
| AT500850B1 (de) | 2007-10-15 |
| US6828124B2 (en) | 2004-12-07 |
| WO2002051238A2 (en) | 2002-07-04 |
| US20030166157A1 (en) | 2003-09-04 |
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