中国比较医学杂志2012，Vol.22Issue(2)：38-42,5.DOI:10.3969/j.issn.1671.7856.2012.02.009
牛γ-干扰素在大肠杆菌中的表达及抗原性鉴定
Expression of Bovine Interferon-γ (BovIFN-γ) in E.coli and Identification of its Antigenicity
刘巧荣 1孙明 1乔明明 1杨璐 1申屠芬琴 1马永缨 1曹振 2田克恭 2陈西钊1
作者信息
- 1. 北京世纪元亨动物防疫技术有限公司,北京100085
- 2. 中国动物疫病预防控制中心,北京100094
- 折叠
摘要
Abstract
Objective To express bovine interferon-^ (BovIFN--/) in E. Coli, and preliminarily identify its biological activity. Methods According to the gene sequence in GenBank to construct the BovIFN--y gene, and use it as the template, to amplify the BovIFN--y gene by polymerase chain reaction (PCR). To insert the BovIFN--y gene into vector PET-28a, transform it into E. Coli BL21 -competent cells, induce it with IPTG to express BovIFN--y, and analyze it by Western blot. Using the NI-NTA affinity chromatography and electroelution to purify the recombination protein, and detect the protein antigenicity by commercial BovIFN-'y kit. Results The recombinant plasmid PET-28a-BIFN--y was successfully constructed, and BovIFN-7 was efficiently expressed in E. Coli. The protein expression accounted for 32% of the total soluble bacterial proteins. The expression products mainly existed in soluble form in the cell lysate supernatant, and the proteion could be discriminated by BovIFN--ymonoclonal antibody. The recombinant protein purified by NI-NTA affinity chromatography had higher antigen activity than that by electroelution. Conclusions The soluble recombinant BovIFN-'yprotein is successfully expressed in E. Coli, and the protein can react with BovIFN--y monoclonal antibody. The purified recombinant BovIFN-"y protein has good antigenicity.关键词
牛γ干扰素/原核表达/抗原性/大肠杆菌Key words
Bovine interferon-'y/ Prokaryotic expression/ Antigenicity/ E. Coli分类
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刘巧荣,孙明,乔明明,杨璐,申屠芬琴,马永缨,曹振,田克恭,陈西钊..牛γ-干扰素在大肠杆菌中的表达及抗原性鉴定[J].中国比较医学杂志,2012,22(2):38-42,5.
