山羊MGAT5基因克隆及其在不同组织和成肌分化过程中的表达研究  

Cloning of goat MGAT5 gene and its expression in different tissues and during myoblastic differentiation process

作　　者：王旭然 李英豪 李红艳 高树新[1] WANG Xuran;LI Yinghao;LI Hongyan;GAO Shuxin(College of Animal Science and Technology,Inner Mongolia University for Nationalities,Tongliao 028000;Tongliao Animal Husbandry Development Center,Tongliao 028001,China;Tongliao Jingyuan Breeding Cattle Co.,Ltd.,Tongliao 028036,China)

机构地区：[1]内蒙古民族大学动物科技学院,内蒙古通辽028000 [2]通辽市畜牧业发展中心,内蒙古通辽028001 [3]通辽京缘种牛繁育有限责任公司,内蒙古通辽028036

出　　处：《黑龙江畜牧兽医》2024年第8期54-59,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：内蒙古自治区“科技兴蒙”行动重点专项“肉牛肉质性状的遗传机理技术研究”(KJXM2020002-05)。

摘　　要：为了探究山羊糖基化酶N-乙酰氨基葡萄糖转移酶(N-acetylglucosaminyl transferase, MGAT) 5基因序列信息及其在不同组织和成肌细胞分化过程中的表达变化,试验以罕山白绒山羊为研究对象,利用PCR技术克隆了山羊MGAT5基因编码序列(coding sequence, CDS),并对其进行了生物信息学分析,同时利用反转录荧光定量PCR(quantitative real-time PCR,qRT-PCR)与Western-blot技术检测MGAT5基因在山羊不同组织(心脏、肺脏、脾脏、肺脏、肾脏、背最长肌)及成肌细胞分化过程中的表达情况。结果表明:山羊MGAT5基因CDS全长为2 220 bp,编码739个氨基酸,与绵羊MGAT5基因亲缘性较高,与小鼠亲缘性较远;MGAT5蛋白的等电点为8.49,为稳定蛋白;MGAT5基因在罕山白绒山羊在心脏、肝脏、脾脏、肺脏、肾脏、背最长肌均有表达,在肾脏中表达量最高,其次为脾脏、肺脏、心脏、肝脏,在背最长肌中表达量最低;MGAT5基因的mRNA和蛋白表达量在山羊成肌细胞分化过程中均呈逐渐降低趋势。说明MGAT5基因的表达可能与成肌细胞分化程度存在负相关关系。The purpose of this study is to explore the sequence information of goat N-acetylglucosaminyl transferase(MGAT)5 gene and its expression differences in different tissues and during myoblastic differentiation process.Hanshan white cashmere goats were selected as study subjects in this experiment,the coding sequence(CDS)region of goat MGAT5 gene was cloned by PCR technique and its bioinformatics was analyzed.Meanwhile,the expression of MGAT5 gene in different tissues(heart,liver,spleen,lungs,kidneys,longissimus dorsi)and during myoblastic differentiation process was examined by quantitative real-time PCR(qRT-PCR)and Western-blot.The results showed that the CDS sequence of MGAT5 gene was 2220 bp,encoding 739 amino acids,which was closely related to MGAT5 gene in sheep,but far related to mouse.The molecular weight of MGAT5 protein was 8.49,and it was a stable protein.The MGAT5 gene was expressed in the heart,liver,spleen,lungs,kidneys,and longissimus dorsi of Hanshan white cashmere goats,with the highest expression level in the kidneys,followed by the spleen,lungs,heart,and liver,and the lowest expression level in longissimus dorsi.In addition,the mRNA and protein expression of MGAT5 decreased during the differentiation of goat myogenic cells,suggesting that MGAT5 gene expression might be negatively correlated with myogenic cell differentiation.

关 键 词：山羊 MGAT5基因 克隆 生物信息学分析 组织表达分析 成肌细胞 分化 

分 类 号：S827[农业科学—畜牧学]