한국양식학회지 (Journal of Aquaculture)

한국양식학회 (The Korean Society of Fisheries and Aquatic Science)
권호 표지

붉바리(Epinephelus akaara)의 성장호르몬 cDNA의 Cloning과 E. coli에서의 발현

Cloning of Growth Hormone Complementary DNA from Red-Spotted Grouper (Epinephelus akaara) and Its Expression in E. coli

  • 강거영 (제주대학교 해양생물공학과) ;
  • 송춘복 (제주대학교 해양생물공학과) ;
  • 이제희 (제주대학교 해양생물공학과)
  • 발행 : 2003.05.01
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초록

붉바리(E. akaara)의 뇌하수체에서 추출한 mRNA로부터 RACE 방법으로 cDNA를 cloning하고 염기서열을 분석하였다. 붉바리의 성장호르몬 cDNA는 5' UTR, open reading frame, 3' UTR이 각각 21 bp, 615 bp, 247 bp인 전체 883 bp로 구성되어있다. Adenylation signal로 작용하는 sequence인 AATAAA는 poly(A)로부터 20 bp upstream에 위치하는 것을 확인하였다. 염기서열을 바탕으로 추정한 성장호르몬은 204개의 아미노산으로 구성된 단백질로서 signal peptide(17 aa)와 mature protein(187 aa)을 포함하고 있으며 mature protein의 분자량은 21.3 kDa으로 나타났다. 이 호르몬은 단백질의 3차 구조에 중요한 이황화 결합을 할 수 있는 4개의 cysteine 잔기와 1개의 N-glycosylation site를 가지고 있었다. 붉바리 성장호르몬의 염기서열은 농어목에 속하는 다른 어류의 성장호르몬 염기서열과의 유사도가 높게 나타났으며 orange-spotted grouper, gilthead seabream과 각각 96.9%, 88.6%의 상동성을 보였다.

We have cloned and sequenced the cDNA encoding growth hormone (GH) from pituitary poly(A)$^{+}$ RNA of red-spotted grouper (Epinephelus akaara). The cDNA of red-spotted grouper GH is 883 base pairs (bp) consisting of 21 bp of 5'untranslated region (UTR), 615 Up of an open reading frame (ORF) and 247 Up of 3'UTR. The polyadenylation signal, AATAAA, was 20 bp upsteam of polyadenylation site. Based on the nucleotide sequences, the deduced putative polypeptide contains 204 amino acids (aa), representing 17 aa of a signal and 187 aa of a mature polypeptide. The putative GH cDNA encodes a polypeptide with four cysteine residues and only one N-gly- cosylation site. Comparative sequence alignment shows that red-spotted grouper GH exhibits high similarity with its corresponding other Perciformes species GH cDNAs.

키워드

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