摘要
通过PCR扩增高致病性禽流感H5N1的HA和HA1基因片段,利用原核表达系统表达重组HA和HA1蛋白,并用SDS-PAGE,Western-blotting和血凝试验进行重组HA蛋白的鉴定和血凝活性鉴定.结果表明,经过HA蛋白胞外段分析、重组载体的酶切鉴定得出的重组HA蛋白,在血凝试验中能与HA特异性抗体结合,并有较高的血凝活性.利用表达载体pET42a表达AIV-H5亚型的HA基因,为进一步利用重组蛋白研制AIV-H5抗体的ELISA检测方法,研制抗AIV-H5亚型的单克隆抗体提供基础.
Amplify the HA and HA1 gene fragment of highly pathogenic avian influenza H5N1 by PCR,express the recombinant HA and HA1 protein by prokaryotic expression system and use the SDS-PAGE,Western-blotting and hemagglutination test for the identification of recombinant HA protein and hemagglutination activity.The result shows that,after analysis of the HA protein ectodomain and identification of the recombinant vector by restriction analysis,the recombinant HA protein can bind to the HA-specific antibody in the hemagglutination test and have a rather high hemagglutination activity.The HA gene of AIV-H5 subtype expressed by expression vector pET42a lays a foundation of the further development of the ELISA method for detecting AIV-H5 antibody by the recombinant protein and the monoclonal antibody against AIV-H5 subtype.
出处
《河南科技学院学报(自然科学版)》
2011年第6期43-48,共6页
Journal of Henan Institute of Science and Technology(Natural Science Edition)
关键词
禽流感
血凝素(HA)蛋白
活性测定
基因表达
avian influenza,hemagglutinin protein,activity determination,gene expression