鲫两种不同生长激素cDNA的分子克隆和分析

MOLECULAR CLONING AND ANALYSIS OF TWO DISTINCT GROWTH HORMONE cDNAs FROMCAR ASSIUS AURATUS AURATUS

  • 摘要: 从鲫脑垂体组织提取总RNA,采用3'RACE-PCR的方法,从单一垂体总RNA中扩增出编码两种不同类型鲫生长激素的cDNA:生长激素Ⅰ(Growth hormone Ⅰ,GH Ⅰ)和生长激素Ⅱ(Growth hormone Ⅱ, GHⅡ).将两种类型的鲫GH cDNA分别克隆到pGEM-T Easy Vector上进行序列测定和分析.克隆的鲫GHⅠ和GHⅡ cDNA均包括编码188个氨基酸残基的GH成熟肽序列和3'端的非翻译区,但不含信号肽序列和5'端非编码区.序列分析结果表明,鲫GHⅠ的碱基序列和推测的氨基酸序列与国外已经发表的金鱼GHⅠ的同源性分别为98.7%和97.9%;GHⅡ的碱基序列和推测的氨基酸序列与金鱼GHⅡ的同源性分别为99.1%和99.5%,虽然同源性较高,但仍具有一定的差异.

     

    Abstract: Growth hormone, a 22-kDa single-chain polypeptid e hormone synthesized and secreted by the anterior pituitary gland, plays an impo rtant role in the regulation of growth, development,metabolism and appetite.It is one of the best-studied pituitary hormones that have been used as models for studying of physiology, nutritional composition, regulating of gene expression, structure-function relationships and gene evolution. In mammals,growth hormone variants arising from differential processing of a single primary RNA transcript have been reported, whereas the growth hormone variants found in teleost appear to be the products of different genes.In previous studies, a few teleost fishes h ave two distinct growth hormones, such as chum salmon(Oncorhynchus keta), ch inook salmon (Oncorhynchus tshawytscha), coho salmon(Oncorhynchus kisutch),Atlantic salmon(Salmo salar), rainbow trout (Oncorhynchus mykiss),At lantic cod(Gadus morhua),Japanese eel(Anguilla japonica),goldfish(Car assius auratus),etc.Growth hormone gene duplication was believed to be the re sult of polyploidization among ancestral aquatic vertebrates. There is a close r elationship betweenCarassius auratus auratusand goldfish, belonging to the same family of Cyprinidae.Carassius auratus auratuswere reported to be pol yploid, so it was possible thatCarassius auratus auratusalso had two disti nct growth hormone genes in a single body.Gene duplication was a rare event. Whe ther there were two formsCarassius auratus auratusgrowth hormone cDNAs enc oding distinct growth hormone was still unknown,it was significative to study th e growth hormone polypeptides ofCarassius auratus auratus.

     

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