CAJ | 학술논문

设计特异性引物,利用RT-PCR方法从牦牛脑垂体中扩增出生长激素基因(YGH)编码区全序列,并进行分子特性分析.结果表明:牦牛生长激素基因编码区全长654 bp,包含一个完整的阅读框,编码217个氨基酸.采用生物信息学方法对分子特性进行分析表明:动物生长激素基因在进化上非常保守,牦牛生长激素基因氨基酸序列与黄牛、山羊、绵羊、马、猪、猫和犬的同源性分别为100 %、99.1 %、98.6 %、88.9 %、88.5 %、88.9 %和89.4 %.牦牛生长激素蛋白有2个明显的疏水区域,存在2个强跨膜区和信号肽,是一种分泌型蛋白质,二级结构主要以α-螺旋和不规则卷曲为主,三级结构包含第28-215位氨基酸残基;牦牛生长激素蛋白有较高的抗原指数和抗原表位,这与YGH编码蛋白具有良好的抗原性相一致.
설계특이성인물,이용RT-PCR방법종모우뇌수체중확증출생장격소기인(YGH)편마구전서렬,병진행분자특성분석.결과표명:모우생장격소기인편마구전장654 bp,포함일개완정적열독광,편마217개안기산.채용생물신식학방법대분자특성진행분석표명:동물생장격소기인재진화상비상보수,모우생장격소기인안기산서렬여황우、산양、면양、마、저、묘화견적동원성분별위100 %、99.1 %、98.6 %、88.9 %、88.5 %、88.9 %화89.4 %.모우생장격소단백유2개명현적소수구역,존재2개강과막구화신호태,시일충분비형단백질,이급결구주요이α-라선화불규칙권곡위주,삼급결구포함제28-215위안기산잔기;모우생장격소단백유교고적항원지수화항원표위,저여YGH편마단백구유량호적항원성상일치.
Growth hormone is a single-chain polypeptide hormone and secreted by anterior pituitary. GH exist in a wide variety of vertebrates, it can promote animal growth and development. The objective of this study was to clone the yak GH gene and do sequence analysis. A pair of specific PCR primers were designed for cloning the coding sequence of GH gene from pituitary tissue in yak. A fragment with 654 bp was got by RT-PCR amplifying, in which included a complete open reading frame and it encoded 217 amino acids(GeneBank accession No. EF154193).Bioinformatics analysis of the molecular characteristics showed that the growth hormone gene was very conservative. Compared nucleotide sequence with cattle, there was only a base difference, which the 607th base was A in yak, but C in cattle, and this difference did not led to amino acid change, and both encoded Arg. The amino acids sequences of yak shared 100 %, 99.1 %, 98.6 %, 88.9 %, 88.5 %, 88.9 % and 89.4 % sequence similarity with cattle, goat, sheep, horse, pig, cat and dog respectively; the protein had two obvious hydrophobic region, existence of two strong transmembrane region and signal peptide. The protein second structure main was α-helix and irregular curly and three-dimensional solution structure composed of 28-215 amino acids. Yak growth hormone protein had higher antigenic index and higher lymphocyte epitopes, which was consistent with of the higher antigenicity of YGH coding protein.