塔什库尔干羊FecB基因非探针法高分辨率熔解曲线分型方法的研究  

Study on non-probe HRM typing of gene of Tashkurgan sheep

作　　者：李浩[1] 郭涛 陈童[1,2] 张璐 张云生 郑新宝 阿依努尔·亚森[1,2] Li Hao;Guo Tao;Chen Tong;Zhang Lu;Zhang Yunsheng;Zhang Xinbao;Aynur Yasin(Institute of Animal Science,Xinjiang Academy of Animal Science,Wulumuqi,830026,Xinjiang;Key Laboratory of Livestock and Poultry Resources(Sheep&Goat)Evaluation and Utilization,Ministry of Agriculture and Rural Affairs,Wulumuqi,830026,Xinjiang;Animal Health Supervision Institute of Xinjiang Uygur Autonomous Region,Wulumuqi,830026,Xinjiang)

机构地区：[1]新疆畜牧科学院,新疆乌鲁木齐830011 [2]农业农村部畜禽资源(羊)评价利用重点实验室,新疆乌鲁木齐830011 [3]新疆维吾尔自治区动物卫生监督所,新疆乌鲁木齐830026

出　　处：《中国畜禽种业》2023年第9期20-25,共6页The Chinese Livestock and Poultry Breeding

基　　金：塔什库尔干羊优质种公羊遗传评估技术研究(KY2022006);外源激素诱导绵羊生产多羔技术研究与示范(2021D03006);新疆地方绵羊与家畜种质资源库及信息共享平台建设(PT2104)。

摘　　要：建立塔什库尔干羊FecB基因非探针法高分辨率熔解曲线分型方法,为塔什库尔干羊多胎类型选育奠定基础。试验选取塔什库尔干羊产双羔(多羔)母羊67只,双羔(多羔)羔羊136只,采集静脉血后,采用非探针法高分辨率熔解曲线对探究塔什库尔干羊FecB基因进行分型的条件。结果表明,115bp扩增引物聚合性以及温度的一致性较好,适合SNP位点的分型,且引物添加量为0.6μL时扩增效率较较好。镁离子浓度在2.4~3.2mmol/L时,扩增曲线和熔解曲线聚合性较好,反应模板浓度在50ng时,扩增曲线、熔解曲线和熔解温度差异曲线较符合塔什库尔干羊FecB基因分型的要求。通过使用优化路线,进行塔什库尔干羊FecB基因分型发现,母羊和羔羊不携带纯合型(BB),携带杂合型(B+)母羊占31.3%,羔羊占25.7%;携带野生型的母羊占68.7%,羔羊占74.3%。。可见,通过建立FecB基因非探针法高分辨率熔解曲线分型方法,筛选较优反应条件,可提高塔什库尔干羊多胎新品系培育的选种效率和选种准确性。The non-probe high-resolution melting curve typing method of FecB gene in Tashkurgan sheep was established,which laid a foundation for the breeding of multiple birth type of Tashkurgan sheep.In this experiment,67 double lambs(polytocous)ewes and 136 double lambs(polytocous)lambs of Tashkurgan sheep were selected.After collecting venous blood,the conditions for genotyping of FecB gene in Tashkurgan sheep were explored by non-probe high-resolution melting curve.The results showed that the 115 bp amplification primer had good polymerization and temperature consistency,which was suitable for the typing of SNP loci,and the amplification efficiency was better when the primer addition amount was 0.6μLL.When the concentration of magnesium ion was 2.4~3.2mmol/L,the amplification curve and melting curve had good polymerization.When the concentration of reaction template was 50ng,the amplification curve,melting curve and melting temperature difference curve were more in line with the requirements of FecB genotyping in Tashkurgan sheep.By using the optimized route,the FecB genotyping of Tashkurgan sheep was found that Ewes and lambs did not carry homozygous type(BB),carrying heterozygous type(B+)ewes accounted for 31.3%,lambs accounted for 25.7%;ewes carrying wild type accounted for 68.7%,and lambs accounted for 74.3%.It can be seen that by establishing a non-probe high-resolution melting curve typing method of FecB gene and screening for better reaction conditions,the selection efficiency and accuracy of the breeding of new multiparous strains of Tashkurgan sheep can be improved.

关 键 词：塔什库尔干羊 FECB基因 非探针法 基因分型 

分 类 号：S826[农业科学—畜牧学]