广灵驴SCD基因克隆、生物信息学及组织差异表达分析  

Cloning, bioinformatics and tissue differential expression analysis of SCD gene in Guangling donkey

作　　者：关家伟 孙瑜彤 邱丽霞 李武峰[1] 杜敏 Guan Jiawei;Sun Yutong;Qiu Lixia;Li Wufeng;Du Min(College of Life Sciences,Shanxi Agricultural University,Taigu 030801,China;Department of Animal Sciences,Wash-ington State University,Pullman 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,华盛顿州普尔曼99164-6310

出　　处：《山西农业大学学报（自然科学版）》2021年第2期65-73,共9页Journal of Shanxi Agricultural University(Natural Science Edition)

基　　金：山西省重点研发计划(指南)项目:国际科技合作方面(201803D421022);山西农业大学横向科技项目(2019HX78)。

摘　　要：[目的]本试验旨在对广灵驴SCD基因CDS区进行生物信息学分析,并检测其在广灵驴组织中的表达,为探究SCD基因在广灵驴脂肪沉积中的作用提供理论参考。[方法]设计SCD基因同源引物,RT-PCR法克隆CDS序列,对SCD基因编码产物进行生物信息学预测,同时使用qRT-PCR法检测广灵驴7个组织中SCD基因的表达量。[结果]广灵驴SCD基因CDS长1080 bp,可编码359个氨基酸,提交到GenBank,获得登录号:MW132164。广灵驴编码序列与马、牛、双峰驼、猪、绵羊、人、小鼠、大鼠的同源性分别为99.4%、89.2%、90.7%、90.1%、89.5%、88.2%、81.2%、82.4%。SCD蛋白分子质量为41.55 ku,等电点为9.29,不稳定指数为45.54,平均疏水指数为-0.146,属于不稳定的碱性亲水蛋白。SCD蛋白既没有信号肽也没有卷曲螺旋,有4个跨膜区,主要定位在内质网。二级、三级结构显示,SCD蛋白主要由α-螺旋(40.67%)和无规则卷曲(40.11%)构成。SCD基因在广灵驴7个组织内都表达,在肝脏、肺脏和皮下脂肪中的表达量最高,心脏和背最长肌最低。[结论]本研究结果可为今后探究SCD基因在广灵驴脂肪沉积中的功能,以及提高广灵驴乳肉品质奠定基础。[Objective]The purpose of this study was to clone the CDS region of SCD gene,to detect its expression in 7 tissues of Guangling donkey using a variety of tools for bioinformatics analysis,and to provide theoretical reference for exploring the role of SCD gene in fat deposition of Guangling donkey.[Methods]Homologous primers of SCD gene was designed,and CDS sequence was amplified with RT-PCR.Functional prediction of SCD gene coding products was made by various bioinformatics methods,and the expression of SCD gene in 7 tissues of Guangling donkey was detected by qRT-PCR.[Results]The length of SCD gene CDS of Guangling donkey was 1080 bp,which encoded 359 amino acids.It was submitted to GenBank,and the accession number was obtained as MW132164.The coding sequence of SCD gene of Guangling donkey showed 99.4%,89.2%,90.7%,90.1%,89.5%,88.2%,81.2%and 82.4%identity with that of Equus caballus,Bos Taurus,Camelus bactrianus,Sus scrofa,Ovis aries,Homo sapiens,Mus musculus and Rattus norvegicus,respectively.SCD protein was an unstable alkaline hydrophilic protein with molecular weight of 41.55 ku,isoelectric point of 9.29,instability index of 45.54 and grand average of hydrophobicity of-0.146.There were four transmembrane regions without signal peptide and coiled helix,which are mainly located in the endoplasmic reticulum.Secondary and tertiary structure prediction showed that SCD protein was mainly composed of alpha helix(40.67%)and random coil(40.11%).SCD gene was expressed in all 7 tissues of Guangling donkey,among which the expression in liver,lungs and subcutaneous fat tissues were the highest,and the expression in heart and longissimus dorsi muscle was the lowest.[Conclusion]The results of this study provided a solid basis for exploring the function of SCD gene in fat deposition of Guangling donkey and improving the quality of Guangling donkey milk and meat in the future.

关 键 词：SCD基因 广灵驴 基因克隆 生物信息学分析 组织表达 

分 类 号：S822[农业科学—畜牧学]