牛皮蝇Hypoderm in B基因的克隆及原核表达载体构建

Cloning and Prokaryotic Expression Vector Construction of Hypodermin B. Gene of H. Bovis

参照牛皮蝇HypoderminB(HB)基因的核苷酸序列,设计一对引物,以牛皮蝇总RNA为模板利用RT-PCR扩增牛皮蝇HB基因,并对扩增基因进行克隆测序。序列分析表明,克隆的牛皮蝇HA基因与GenBank中已经登录的核苷酸和氨基酸序列的同源性分别为97.2%和99.3%。同时,将该基因与原核表达载体PGEX-4T-1连接,构建并获得阳性重组表达载体。

Hypodermin B. gene of H. Bovis was firstly amplified by RT-PCR with the primers designed on the nucleotide sequence of Hypodermin B. gene and the total RNA of H. Bovis, then the Hypodermin B. gene was cloned and sequenced. Results showed that the homologies of the Cloned Hypodermin B. gene nucleotide and amino acid sequences and the relative sequences in GenBank were 97.2 percent and 99.3 percent, respectively. The positive recombined expression vector of Hypodermin B. gene was finally constructed with the prokaryotic expression vector of PGEX-4T-1 and identified.