猪γ-干扰素全基因克隆及原核表达  

Cloning and Prokaryotic Expression of Interferon Gamma Full Gene in Pig

作　　者：马超英 

机构地区：[1]青海省海西州乌兰县铜普兽医站,青海乌兰817000

出　　处：《中国畜牧兽医》2012年第10期91-94,共4页China Animal Husbandry & Veterinary Medicine

摘　　要：利用RT-PCR技术,从ConA活化的猪外周血淋巴细胞中扩增出γ-干扰素(interferon-γ,IFN-γ)cDNA,克隆到pMD18-T载体中进行测序后,与原核表达载体pET32a(+)重组,表达含His×6的IFN-γ重组蛋白;测序结果与GenBank中已发表的序列进行比对,核苷酸同源性在98.6%～100.0%之间,氨基酸同源性在96.4%～99.4%;经SDS-PAGE和Western blotting分析结果表明,该重组蛋白分子质量约为27ku,主要存在于包涵体中,且具有良好的免疫学活性,为进行γ-干扰素深入的研究奠定了基础。To amplify pig interferon-γ（pIFN -γ） cDNA from ConA activated peripheral-blood lymphocyte by RT PCR, then was cloned into pMD18-T and sequenced, and recombined with pET32a（＋） vector, the target protein His× 6 IFN-γ was in duced by IPTG;the sequence analysis showed that the nucleotide sequences of the IFN-γ in the study shared 98.6% to 100.0 homology with that of pig published in GenBank, and the identity of amino acid was 96.4% to 99.4%. This recombinant pro- rein, a molecular weight of about 27 ku and mainly presenting in inclusion bodies, was approved to have immunological activity by Western blotting analysis and SDS-PAGE. It laid a foundation for further research of IFN-γ.

关 键 词：基因克隆 原核表达 IFN-Γ基因 猪 

分 类 号：Q78[生物学—分子生物学]