天祝白牦牛论文天祝白牦牛FSHFSHR基因多态性分析与双胎性状的关联研究.doc

天祝白牦牛论文：天祝白牦牛FSH、FSHR基因多态性分析与双胎性状的关联研究 【中文摘要】本实验以天祝白牦牛为研究对象,采用PCR-HRM和PCR-SSCP方法对FSHβ基因的5′端、FSHR基因5′端和第10外显子的多态性进行分析,旨在探索天祝白牦牛双胎牛的分子标记,为今后天祝白牦牛双胎选育提供分子育种理论。研究结果如下:(1)FSHβ基因5′端多态性及序列分析,天祝白牦牛在FSHβ基因5′端的2对引物P1和P2扩增片段,经HRM扫描发现,P2片段具有突变位点。再经SSCP检测,在天祝白牦牛上检测到AA、AB型两种基因型,群体多态信息含量(PIC)小于0.25。测序结果显示,P2片段中AB基因型个体在FSHβ基因5′端的-2009bp碱基处发生了突变,碱基突变为C变为T,氨基酸由Leu变为Pro。(2) FSHR基因5′端多态性及序列分析,对于天祝白牦牛FSHR 5′端扩增片段P3和P4扩增片段中,经HRM扫描发现,P4片段没有多态性,P3片段在天祝白牦牛上检测到了CC、CD两种基因型。群体多态信息含量(PIC)小于0.25。出现了正常型和突变型个体,突变个体在FSHR基因5′端-1195bp处发生了突变,碱基由G变为A,氨基酸由Met变为Val。(3) FSHR第10外显子P5,P 6扩增片段在被检测的天祝白牦牛中均未检测到多态性。这与其它双胎牛、多胎羊等报道有差异。(4) FSH和FSHR基因多态性与双胎性状的关系,经过对本实验设计的6对引物扩增片段比对分析发现,引物FSHβP2扩增片段(-2009bp处),FSHR基因P3片段(-1195bp处)在天祝白牦牛单胎群体和双胎群体中,双胎母牛的突变率显著高于单胎母牛的突变率,说明FSH和FSHR基因很可能是控制天祝白牦牛双胎性状的主效基因,同时也证明了PCR-HRM和PCR-SSCP分子标记进行辅助标记选择是可行的。这为天祝白牦牛双胎性状的选育提供了分子育种基础,此前一直认为牦牛属于单胎动物,未曾有过双胎的报道。 【英文摘要】The polymorphisms of 5′flanking region of FSHβand FSHR gene together with exon10 of FSHR gene were detected in Tianzhu white yak with PCR-HRM and PCR-SSCP , And the association between the polymorphisms and the Twinning traits were analyzed. The aim was to choose perfect markers which can provide theoretical base for breeding in Tianzhu white yak. The results were as follows:(1)The polymorphisms and analysis of 5′flanking region of FSHβ: Two primers P1 and P2 were designed to amplify the 5′flanking region of FSHβgene, after scanning by HRM, PCR product amplified by primer P2 were found polymorphism. When detected by SSCP, two genotypes AA and AB were found, the polymorphism information content (PIC) was low (PIC0.25), showed that there was Less genetic variation. The sequencing results showed that this polymorphism was caused by mutation C to T in position 2009, this mutation caused amino acid Leu to Pro.(2) The polymorphisms and analysis of 5′flanking region of FSHR:Two primers P3 and P4were designed to amplify the 5′flanking region of FSHR, after scanning by HRM, P