黄牛MEF2A基因克隆及真核表达载体构建

摘要

The study aimed to clone cattle MEF2A gene and construct its eukaryotic expression vector, and provide the genetic basis for the efficient selection of Chinese cattle and the foundation of molecular marker database as well as the germplasm conservation and application. The MEF2A gene of cattle was cloned into eukaryotic expression vector, constructing recombinant plasmid of pEGFP-C1-MEF2A. Bioinformatical tools were abopted to analyz characteristics of cattle MEF2A gene and the protein. The cattle MEF2A gene, a 1 497 bp length coding region (CDS), coded 498 amino acids. The prediction demonstrated that MEF2A was composed of a MADS-box domain(No. 2-56 amino acid) , a MEF2 domain (No. 57-77 amin acid). There was no functional domain was founden in c-terminal. An eukaryotic espression vector pEGFP-C1-MEF2A was constructed successfully.%旨在克隆MEF2A基因和构建其真核表达载体,为黄牛种质资源保存利用以及肉牛转基因育种和产业化提供基因资源和育种素材.本研究通过RT-PCR克隆黄牛MEF2A基因CDS区,并将其插入到质粒载体pEGFP-C1的多克隆位点中,构建真核表达载体pEGFP-C1-MEF2A.同时应用生物学软件分析MEF2A基因及其编码蛋白的生物学特性,了解其复杂的调控机能.结果,MEF2A基因的CDS区全长1494 bp,编码498个氨基酸残基.生物信息学分析表明,N端第2～56位氨基酸肽段为MADSbox结构域,第57～77位氨基酸肽段为MEF2结构域；C端没有明显的功能域.成功的构建了含有牛MEF2A基因的真核表达载体pEGFP-C1-MEF2A.