中国畜牧兽医2013，Vol.40Issue(4)：40-45,6.
犬瘟热病毒H蛋白的原核表达及免疫原性的初步鉴定
Prokaryotic Expression and Idenfication of the Preliminary Immunogenicity of the H Protein of Canine Distemper Virus
摘要
Abstract
The assay was aimed to highly express the H gene of canine distemper virus in E. coli and analyze the immunogenicity of the products. The fragment with main epitope sequences, amplified from Ondetstepoort vaccine strain via reverse transcription-polymerase chain reaction, was cloned into vector pET28b and expressed through IPTG introduction after transformed into Ro-setta(TM) strain. The product, after purification, was identified by SDS-PAGE, western blotting and indirect enzyme linked im-munosorbent assay. After that, the antibody titers of mice immunized with the purified H proteins were determined with neutralization test. The 1113 bp fragment of interest was specifically amplified by RT-PCR and expressed with the introduction of 1.0 mmol/L IPTG at 37 ℃. The protein expressed, identified by SDS-PAGE and Western blotting with a size of 42. 38 ku, could be recognized by anti-CDV antibody without nonspecific reaction. In the animal experiment, the serum antibody titer was determined to be 2-3.3 by neutralization test. The target protein was expressed accurately and specifically, showing a good im-munogenicity and antigenicity according to the animal experiment, and therefore could be employed as an antigen candidate for the Real-time examination of animal immune condition, which laid the foundations for the development of new-type canine distemper vaccines and ELISA kit for detecting the anti-CDV antibody.关键词
犬瘟热病毒/H基因/原核表达/Western blotting/活性Key words
canine distemper virus/haemagglutinin gene/prokaryotic expression/Western blotting/ activity分类
生物科学引用本文复制引用
袁颖烁,宋菲菲,张乐萃,扈荣良..犬瘟热病毒H蛋白的原核表达及免疫原性的初步鉴定[J].中国畜牧兽医,2013,40(4):40-45,6.基金项目
国家公益性行业(农业)科研专项经费资助(201203056). （农业）
