• 제목/요약/키워드: gilthead seabream

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Expression of GiIthead Seabream (Sparus aurata) Growth Hormone in Escherichia coli Using Alginate Lyase Gene Promoter of Pseudomonas sp.

  • Lee Jong-Hee;Choi Sun-Young;Lee Sang-Bong;Jin Cheal-Ho;Huh Sung-Hoi;Kong In-Soo
    • Fisheries and Aquatic Sciences
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    • 제2권1호
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    • pp.93-97
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    • 1999
  • The promoter region of alginate lyase gene (aly) from Pseudomonas sp. W7 was used for the high expression of gilthead seabream (Sparus aurata) growth hormone (GH) gene in Esherichia coli. PCR product encoding the premature segment of the growth hormone. was cloned to the downstream of aly promoter. GH was overexpressed With 46 ammo acid of alginate lyase as fusion protein. GH was immunoreactive and production of GH was repressed with supplementation of $0.4\%$ glucose into culture media.

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Molecular Cloning of Red Seabream, Pagrus major Somatolactin cDNA and Its Expression in Escherichia coli

  • Munasinghe, Helani;Koh, Soon-Mi;Lee, Jehee
    • 한국양식학회지
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    • 제16권3호
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    • pp.165-170
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    • 2003
  • Isolation, cloning and sequencing of red seabream (Pagrus major) somatolactin (rsbSL) cDNA from pituitary gland revealed an open reading frame of 693 bp coding for a pre-growth hormone of 231 amino acids with a 22 amino acid putative signal peptide. Deduced amino acid sequence showed that there was one possible N-glycosylation site at Asn$^{145}$ and seven Cys residues (Cys$_{29}$ , Cys$^{39}$ , Cys$^{66}$ , Cys$^{89}$ , Cys$^{205}$ , Cys$^{222}$ , Cys$^{230}$ ). Except Cys$^{66}$ , others may be involved in disulfide bond formation. The rsbSL presented a 93% amino acid sequence identity with the SL of gilthead seabream (Sparus aurata) and contained the conserved hormone domain region. Expression of rsbSL in E. coli (BL2l) cells and gel analysis revealed a higher molecular weight for rsbSL than expected theoretically, implying posttranslational modifications.

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;송춘복;이제희
    • 한국양식학회지
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    • 제16권2호
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    • pp.110-117
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    • 2003
  • 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.