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藏山羊瓣胃溶菌酶基因TGOLyz的克隆及组织表达分析

DOI: 10.3969/j.issn.1000-7091.2013.03.007, PP. 35-42

Keywords: 藏山羊,TGOLyz基因,克隆,定量表达,生物信息学

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Abstract:

基于物种同源性,应用PCR和RT-PCR技术,克隆得到藏山羊瓣胃溶菌酶基因cDNA编码序列,并命名为TGOLyz,应用生物信息学软件对TGOLyz基因核苷酸序列及预测的蛋白序列进行了分析,并预测该蛋白的三级结构;应用qRT-PCR技术检测了TGOLyz在5个组织中的定量表达。结果表明,藏山羊瓣胃溶菌酶基因TGOLyzcDNA编码区长444bp(AccessionNo.KC558501),编码147个氨基酸,分子量为16.29kDa,等电点为6.08。同源性对比显示,TGOLyz与牛胃1(Cowstomach1)的同源性最高,达95.24%,与牦牛胃(Yakstomach)的同源性最低,仅为67.61%;系统进化树显示,该基因属于c型溶菌酶(c-typelysozyme)家族;qRT-PCR分析表明,TGOLyz基因在5个组织中均有表达,在瓣胃中表达量最高。这为进一步深入研究TGOLyz的抗菌及消化功能的分子调控机制提供了理论基础。

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