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虹鳟生长激素:cDNA的分子克隆及在大肠杆菌中的表达

Rainbow trout growth hormone: molecular cloning of cDNA and expression in Escherichia coli.

作者信息

Agellon L B, Chen T T

出版信息

DNA. 1986 Dec;5(6):463-71. doi: 10.1089/dna.1.1986.5.463.

Abstract

We have isolated several recombinant clones carrying the complementary DNA (cDNA) sequence of the rainbow trout (rt) Salmo gairdneri growth hormone (GH) mRNA by immunoblot screening using an antiserum to chum salmon (Oncorhyncus keta) GH. The nucleotide sequence of one of the rtGH cDNA clones (pAF51) was determined. The rt cDNA sequence in pAF51 encodes a hybrid polypeptide of 199 amino acid residues containing 9 amino acid residues of the bacterial beta-galactosidase, one residue from the codon at the junction of the beta-galactosidase gene, and the rtGH cDNA sequence, an additional residue from the presumptive signal peptide of the pre-rtGH and the entire sequence of the mature rtGH (188 amino acid residues). Pairwise matrix comparisons of the hydropathy profiles of bovine, human, rat, and rainbow trout GH polypeptides indicate that regions of similarity exist between the rtGH and mammalian GH. In particular, there are two major regions of similarity found near the amino-terminal region and at the carboxy-terminal region. These regions correspond to hydrophilic domains of the GH molecules. The possible significance of these domains is discussed.

摘要

我们通过使用大麻哈鱼(Oncorhyncus keta)生长激素抗血清进行免疫印迹筛选,分离出了几个携带虹鳟(rt)鲑鱼生长激素(GH)mRNA互补DNA(cDNA)序列的重组克隆。测定了其中一个rtGH cDNA克隆(pAF51)的核苷酸序列。pAF51中的rt cDNA序列编码一个由199个氨基酸残基组成的杂合多肽,其中包含9个细菌β-半乳糖苷酶的氨基酸残基、一个来自β-半乳糖苷酶基因连接处密码子的残基、rtGH cDNA序列、一个来自前rtGH推定信号肽的额外残基以及成熟rtGH的完整序列(188个氨基酸残基)。对牛、人、大鼠和虹鳟GH多肽的亲水性图谱进行成对矩阵比较表明,rtGH与哺乳动物GH之间存在相似区域。特别是,在氨基末端区域附近和羧基末端区域发现了两个主要的相似区域。这些区域对应于GH分子的亲水区。讨论了这些结构域的可能意义。

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