皮蝇Hypodermin C基因的克隆及重组表达载体的构建  

Cloning and construction of recombinant expression vector of Hypodermin C

作　　者：杨东英[1] 罗建勋[2] 唐立刚 关贵全[2] 马米玲[2] 刘志杰[2] 刘爱红[2] 党志胜[2] 王艳华[2] 殷宏[2] 欧阳五庆[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]中国农业科学院兰州兽医研究所,甘肃兰州730046 [3]山东省烟台市芝罘区畜牧公司,山东烟台264000

出　　处：《中国兽医科技》2003年第5期3-6,共4页Chinese Journal of Veterinary Science and Technology

摘　　要：参照牛皮蝇HypoderminC(HC)基因的核苷酸序列 ,设计了 1对引物 ,以牛皮蝇总RNA为模板 ,用RT PCR扩增了牛皮蝇HC基因 ,将扩增基因进行克隆测序。测序结果表明 ,所获得基因与已知序列同源性达 99.4 5 %。同时 ,将该基因与表达载体 pGEX 4T 1连接 ,构建并获得阳性重组表达载体。Total RNA was extracted from Hypoderma, then mRNA was isolated by using oligo(dT) as a probe. The HC gene specific primers were devised by GOLDkey software. A 713 bp specific fragment was amplified by RT PCR and ligated into pGEM T Easy vector. It was identified by restriction endonuclease analysis and PCR and sequencing that this fragment contained the complete open reading frame (ORF) of the HC gene. In comparison with GenBank data, the homologies of the nucleotide sequence and amino acid sequence are 99.45% and 98.36%, respectively. After the HC T was restrictively digested, the interested gene HC was subcloned into the prokaryotic expression vector pGEX 4T 1. Positive clones were selected by restriction endonuclease analysis and PCR identification.

关 键 词：皮蝇 HypoderminC基因 基因克隆 表达载体 牛皮 蝇蛆病 

分 类 号：S858.23[农业科学—临床兽医学] S852.743[农业科学—兽医学]