摘要
构建甲型流感病毒NP基因重组质粒,制备NP蛋白,建立甲型流感病毒胶体金检测方法。设计引物,扩增人工合成的甲型流感病毒株A(H5 N1)(GenBank登录号AY585439)NP DNA基因,构建pET-30 Xa/LIC-NP重组质粒,转化入原核表达系统E.coliBL21 Gold中进行表达和纯化。结果制备了纯度高达900 mL/L的甲型流感病毒NP融合蛋白,并初步建立了金标检测方法。
To construct the recombinant influenza A virus NP plasmid,prepare the NP protein,and establish the influenza A virus colloidal gold assay methods,the full-length synthesized DNA sequence of influenza virus A NP（H5N1）（GenBank accession number AY585439） was amplified by the designed primers,and the pET-30 Xa/LIC-NP recombinant plasmid was constructed.Further,the recombinant plasmid was transformed into the prokaryotic expression bacteria E.coli BL21 Gold,then the recombinant NP protein was expressed and purified.The recombinant soluble protein of influenza A virus NP was successfully obtained and the purity was up to 900 mL/L.Especially,the initial gold immuno-chromatography assay method was established.
出处
《动物医学进展》
CSCD
北大核心
2010年第10期19-23,共5页
Progress In Veterinary Medicine
基金
河南省杰出人才创新基金(074200510003)资助
关键词
甲型流感病毒
原核表达
NP蛋白
胶体金检测
Influenza A virus
prokaryotic expression
nucleoprotein protein
gold immuno-chromatography assay
