中国畜牧兽医2017，Vol.44Issue(8)：2369-2377,9.DOI:10.16431/j.cnki.1671-7236.2017.08.022
水牛Smad4基因的克隆及其真核表达载体的构建
Cloning and Construction of Eukaryotic Expression Vector of Buffalo Smad4 Gene
摘要
Abstract
This study was aimed to elucidate the molecular mecbanisms of Smad4 gene on granulose cells proliferation and embryonic development in buffalo.The Smad4 gene was amplified by RT-PCR,analyzed by bioinformatics,studied with eukaryotic vector construction,and used liposome transfection skill to transfect into the buffalo granulose cells.The results showed that Smad4 gene was cloned,the coding region was 1 662 bp,and encoded 553 amino acids.BLAST analysis showed that the buffalo Smad4 gene shared 99％of similar nucleotide sequence with Bos taurus,and shared 98％,96％,96％and 95％of similar nucleotide sequence with Ovis aries,Sus scrofa,Equues caballus and Homo sapiens,respectively.Phylogenetic tree analysis showed that Smad4 gene was highly conserved in different species.The buffalo pEGFP-N1-Smad4 eukaryotic expression vector was successfully constructed,and transferred into the buffalo granulose cells,and the Smad4-EGFP fusion protein was detected in the cells.The results suggested that the success cloning and construction vector of buffalo Smad4 gene laid the foundation for the research of Smad4 gene on embryo development.关键词
水牛/Smad4基因/克隆/真核表达载体Key words
buffalo/Smad4 gene/cloning/eukaryotic expression vector分类
生物科学引用本文复制引用
鄢胜飞,张秀芳,黄玥萌,郑海英,杨春艳,覃广胜,黄加祥,尚江华..水牛Smad4基因的克隆及其真核表达载体的构建[J].中国畜牧兽医,2017,44(8):2369-2377,9.基金项目
广西科技计划项目(桂科AB16380040) （桂科AB16380040）
广西自然科学基金(2014GXNSFAA118116) （2014GXNSFAA118116）
广西水产畜牧兽医局科技项目(桂渔牧科201633052) （桂渔牧科201633052）
广西水牛研究所基本科研业务费(水牛基1404004、160204、1705001) （水牛基1404004、160204、1705001）
