广灵驴CAST基因的克隆与表达  

Cloning and expression analysis of CAST gene in Guangling donkey

作　　者：李武峰[1] 赵婧微 孙瑜彤 杜敏 LI Wufeng;ZHAO Jingwei;SUN Yutong;DU Min(College of Life Sciences,Shanxi Agricultural University,Taigu,Shanxi 030801,China;Department of Animal Sciences,Washington State University,Pullman,WA 99164-6310,USA)

机构地区：[1]山西农业大学生命科学学院,山西太谷030801 [2]美国华盛顿州立大学动物科学系,美国华盛顿州普尔曼99164-6310

出　　处：《福建农林大学学报（自然科学版）》2020年第5期652-659,共8页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：山西省重点研发计划(国际科技合作)项目(201803D421022).

摘　　要：对广灵驴钙蛋白酶抑制(CAST)基因进行克隆及序列分析,并用实时荧光定量PCR技术对其在心、肝、脾、肺、肾和背最长肌等6种组织中的表达水平进行分析,探究CAST基因对驴肉品质的影响.结果表明,广灵驴CAST基因的CDS编码区全长2427 bp,共编码808个氨基酸,将测序序列提交至NCBI,得到的GenBank登录号为MN231002.广灵驴CAST蛋白的氨基酸序列与马、牛、人、猪、山羊、绵羊、小鼠、鸡的同源性分别为98.6%、77.1%、70.8%、77.5%、70.1%、60.9%、65.1%和41.5%;CAST基因在广灵驴6种组织中均存在表达,其中,表达量最高的组织为背最长肌.本试验成功获得了广灵驴的CAST基因,并发现其在背最长肌中高度表达,为进一步研究CAST基因在肌肉嫩度方面的表达调控功能提供参考.In order to explore the effect of calpastatin(CAST)gene on meat quality of Guangling donkey,the CAST gene was cloned and sequenced,and the expressional levels of CAST gene in different tissues of heart,liver,spleen,lung,kidney and longissimus dorsi muscle were analyzed by real-time PCR.The results showed that the CAST gene fragment included 2427 bp,encoded 808 amino acids,whose accession number is MN231002 in GenBank.The amino acid sequence of Guangling donkey CAST protein was 98.6%,77.1%,70.8%,77.5%,70.1%,60.9%,65.1%and 41.5%homologous to horse,cattle,human,swine,goat,sheep,mouse and chicken.The CAST gene was expressed in all 6 tissues,whose expression level was maximized in longissimus dorsi and followed by heart.The study provides theoretical basis for further study of the expression and regulatory function of CAST gene in muscle tenderness.

关 键 词：广灵驴 钙蛋白酶抑制(CAST)基因 基因克隆 组织表达 

分 类 号：S822[农业科学—畜牧学]