皖南黄兔IGF1基因扩增及其多态性分析  

Amplification and Polymorphism Analysis of IGF1 Gene of Wannan Yellow Rabbit

作　　者：黄冬维[1] 汪勇 杨普[1] 程广龙[1] 汪善义 吴东[1] 赵小伟[1] 杨永新[1] 王小飞[1] 赵辉玲[1] 

机构地区：[1]安徽省农业科学院,合肥230031 [2]安徽省义华农牧科技有限公司,池州245106

出　　处：《中国草食动物科学》2016年第6期6-10,共5页China Herbivore Science

基　　金：安微省科技攻关项目(1501031098);国家兔产业技术体系(CARS-44-A-3)

摘　　要：胰岛素样生长因子1（IGF1）是动物生长发育等生命过程中的重要调节因子,目前关于家兔IGF1基因序列扩增和多态性的研究还很少。该研究以安徽皖南黄兔为试验动物,通过PCR方法扩增其IGF1基因编码区全序列,利用混合DNA池测序和生物信息学方法分析了IGF1基因扩增区域的遗传变异特性。结果获得皖南黄兔IGF1基因包含第1-5外显子的基因组序列5条,长度分别为316、657、691、757和403 bp,与种子序列一致性分别为100%、99.5%、99.9%、99.7%和99.8%;5个外显子组成432 bp的CDS序列,编码143个氨基酸残基组成的蛋白质。测序和生物信息学分析结果则表明,在皖南黄兔IGF1基因内含子1中存在3个多态位点（3056G〉A、3107T〉C和3164 G〉A）,3个位点并不位于剪接位点,但分别位于MZF1、ATF4、CEBPA、HLF和NKX2-8顺式作用元件位置,因而可能影响基因表达。该研究成功扩增获得皖南黄兔IGF1基因全部5个外显子序列,拼接获得基因编码区全序列,并利用混合DNA池测序法发现了内含子3个新的SNP位点,研究结果为揭示肉兔IGF1基因功能奠定了基础。Insulin-like growth factor 1（IGF1）is one of important regulators in animal growth and development,while at present,it remains unclear for the sequence structure and its polymorphism of IGF1 gene in rabbit. In this study,whole genomic sequences of coding region of rabbit IGF1 gene were amplified successfully using PCR method and their polymorphismes were analyzed by ways of pool DNA sequencing and bioinformatics. The result showed that five sequences compromising exon 1 to 5 were 316 bp,657 bp,691 bp,757 bp and 403 bp in lengh,respectively,with highly similarity to the temple sequences（100%,99.5%,99.9%,99.7%and 99.8% respectively）;the five exons constituted the CDS（coding sequence）of 432 bp in lengh,which could be translated to143 amino acid residues of IGF1. Additionally,there were three polymorphic sites（3056G〉A,3107T〉C and 3164 G〉A）in intron1,which did not locate in splicing sites but in several cis-acting elements such as MZF1,ATF4,CEBPA,HLF and NKX2-8,with possibility of regulating gene expression. To sum up,the five exons sequences of IGF1 gene of Wannan yellow rabbit and its coding region were acquired,and so did the three new polymorphic sites of partial intron 1. The results of this study can lay foundations for revealing function of IGF1 gene in meat rabbit.

关 键 词：皖南黄兔 IGF1基因 扩增 多态性 

分 类 号：S829.2[农业科学—畜牧学]