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Nucleotide sequence of a cloned cDNA for proopiomelanocortin precursor of chum salmon Onchorynchus keta.

机译:鲑鱼Onchorynchus keta的proopiomelanocortin前体的克隆cDNA的核苷酸序列。

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摘要

We have isolated a cDNA clone encoding salmon proopiomelanocortin precursor. Polyadenylated RNA was isolated from pituitary neurointermediate lobes and used to construct a cDNA library. The library was screened with 17 mer of oligodeoxyribonucleotides specific for the hexapeptide sequence in salmon beta-endorphin I, Phe-Met-Lys-Pro-Tyr-Thr at positions 4-9 excluding the third nucleotide. One positive clone, pSSM17 containing an insert of 1303 base pairs (bp) was characterized. Sequence determination revealed that it possessed sequences covering the entire regions encoding ACTH and beta-lipotropin and that the mRNA had the same overall organization as those of other mammalian species, i.e., the following peptide hormones were arranged in order from 5' upstream, ACTH including alpha-melanotropin and corticotropin-like intermediate lobe peptide, beta-lipotropin including gamma-lipotropin, beta-melanotropin and beta-endorphin. Amino acid sequences for putative salmon ACTH, beta-, and gamma-lipotropin were predicted. Comparison of the salmon mRNA sequence with those of mammals showed that the regions of alpha- and beta-MSH are relatively homologous, but other regions are much less so, especially in the 3' nontranslated region where it is much longer and completely heterologous.
机译:我们已经分离出编码鲑鱼proopiomelanocortin前体的cDNA克隆。从垂体神经中间叶中分离出聚腺苷酸化的RNA,并用于构建cDNA文库。用对鲑鱼β-内啡肽I,Phe-Met-Lys-Pro-Tyr-Thr的六肽序列具有特异性的六聚体的17 mer寡脱氧核糖核苷酸筛选该文库,但不包括第三个核苷酸。鉴定了一个阳性克隆pSSM17,其中包含1303个碱基对(bp)的插入片段。序列测定表明,它具有覆盖编码ACTH和β-脂蛋白的整个区域的序列,并且该mRNA具有与其他哺乳动物物种相同的整体组织,即,以下肽激素从5'上游开始按顺序排列,ACTH包括α-促黑素和促肾上腺素样的中间叶肽,β-促脂蛋白,包括γ-促脂素,β-促黑素和β-内啡肽。预测鲑鱼ACTH,β-和γ-脂蛋白的氨基酸序列。鲑鱼mRNA序列与哺乳动物序列的比较表明,α-和β-MSH的区域相对同源,但其他区域的同源性要低得多,尤其是在更长且完全异源的3'非翻译区域。

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