人博卡病毒相关基因的克隆与表达

Human bocavirus associated gene cloning and expression

目的克隆临床标本中人博卡病毒相关功能基因,进而在原核表达体系中表达博卡病毒相关的蛋白。方法采用分子生物学技术构建重组质粒PGEX-20T-VP、PGEX-20T-NP和PGEX-20T-NS,分别对其进行PCR、双酶切和测序鉴定,然后将已测序鉴定的包含3种重组质粒的工程菌,转化至大肠杆菌BL21菌中表达博卡病毒VP、NP和NS部分基因的相关蛋白。结果成功的构建了人博卡病毒3种相关功能基因重组质粒,并在大肠杆菌中表达了VP、NP和NS部分基因的相关蛋白。结论原核体系中表达出来的博卡病毒VP、NP和NS部分基因的相关蛋白将为下一步抗原纯化、单抗制备及博卡病毒检测奠定基础。

Objective To clone the genes associated with function researchment on human bocavirus,and to express the proteins about human bocavirus were in prokaryotic expressing system.Methods Recombinant plasmid PGEX-20T-VP,PGEX-20T-NP and PGEX-20T-NS were constructed by molecular biological methods,which were identified by PCR,restriction enzyme digestion and sequence analysis.Those three recombinant plasmids,identified by sequencing,were transformed into E.coli BL21 and expressed human bocavirus associated protein on VP、NP and NS partial gene.Results Three recombinant plasmids were constructed successfully,whereafter human bocavirus associated protein on VP、NP and NS partial gene were expressed.Conclusion These proteins being expressed in prokaryotic expressing system will set up the foundations for antigen purification,monoclonal antibody preparation and human bocavirus detection in next step.