浙东白鹅IGF1基因克隆、组织表达及与体重性状关联分析

Cloning,Tissue Expression and Association Analysis with Body Weight of IGF1 Gene in Zhedong White Goose

为研究胰岛素样生长因子1(Insulin-like growth factor 1,IGF1)基因序列特征,分析其在浙东白鹅各组织中的表达分布及与体重性状的相关性,研究以浙东白鹅肝脏组织为材料,采用RT-PCR技术克隆获得鹅IGF1基因cDNA序列并进行生物信息学预测分析;通过qPCR技术检测IGF1基因在浙东白鹅不同发育阶段各组织中的表达分布;利用PCR产物直接测序和PCR-RFLP技术筛选浙东白鹅IGF1基因的单核苷酸多态性,并分析其与体重性状的相关性。结果显示:浙东白鹅IGF1基因cDNA长633 bp,其开放阅读框长462 bp,可编码153个氨基酸残基,与绿头野鸭IGF1完全同源;IGF1基因在雌性鹅胚(28胚龄)和青年母鹅(70日龄)的肝脏和腿肌以及青年母鹅(70日龄)卵巢组织中高表达;在其内含子2上检测到两个SNP位点(g.5317A>C和g.40793G>C),两突变位点与浙东白鹅不同周龄(2、4、6、8和10周龄)体重性状均无显著相关(P>0.05)。结果表明,IGF1基因在浙东白鹅母鹅腿肌和卵巢组织中高表达,可能在鹅肌肉发育和卵巢发育中起关键作用。

The aim of this study was to analyze the insulin-like growth factor 1(IGF1)gene sequence feature,the expression distribution in various tissues and its correlation with body weight traits of Zhedong White goose.The cDNA sequence of IGF1 gene was cloned by RT-PCR and analyzed by bioinformatics.The tissue expression profile of IGF1 gene at different developmen⁃tal stages of Zhedong White goose was examined by qPCR.Single nucleotide polymorphisms in Zhedong White goose IGF1 gene were screened by direct sequencing of PCR products and PCR-RFLP technique,and its association with body weight traits was analyzed.The results showed that Zhedong White goose IGF1 gene cDNA was 633 bp,which contained one open reading frame of 462 bp encoding a protein of 153 amino acid residues.The deduced IGF1 amino acid shared completely homologous to that of duck.The IGF1 gene was highly expressed in liver and leg muscle tissues of the female goose embryos(28 days of embryonic age)and young female geese(70 days of age),as well as in the ovary of the young female Zhedong White goose(70 days of age).Two single nucleotide polymorphism sites(g.5317A>C and g.40793G>C)were detected in the intron 2 region,the two mutations were not significantly associated with the body weight traits at different weeks of age(2,4,6,8 and 10 weeks of age)in Zhedong White geese(P>0.05).The results indicated that IGF1 gene was highly expressed in the leg muscle and ovary tissues of Zhedong White goose,wich proved that IGF1 gene might play a key role in goose muscle development and ovary development.