鹅细小病毒YBLJ株VP3基因的真核表达  

Eukaryotic Expression of VP3 Gene of Goose Parvovirus YBLJ Strain

作　　者：张颖[1] 鲁承[1] 赵文婧[1] 陈海迪[1] 高旭[1] 

机构地区：[1]延边大学农学院动物医学系,吉林延吉133000

出　　处：《动物医学进展》2012年第12期23-25,共3页Progress In Veterinary Medicine

基　　金：吉林省自然科学基金项目(201215230);延边大学科技发展计划项目(601010002)

摘　　要：为了研究鹅细小病毒(GPV)YBLJ株VP3基因的生物学特性,根据已克隆的GPV YBLJ株VP3基因序列(GenBank:JN836326),设计并合成一对特异性引物,经PCR扩增、克隆、转化、酶切与亚克隆,构建真核表达载体pcDNA-VP3,将鉴定正确的质粒转染至Vero细胞,经RT-PCR检测VP3基因转录水平,通过间接免疫荧光试验(IFA)观察VP3基因表达水平。结果显示,克隆的VP3基因全长约1 605bp,构建了真核表达载体pcDNA-VP3,经RT-PCR对转染细胞总RNA扩增得到特异性目的片段,IFA检测显示VP3基因在Vero细胞中获得了稳定表达。本研究为GPV核酸疫苗的研制奠定了基础。To investigate the biological characteristics of YBLJ VP3 gene of goose parvovirus （GPV）, according to the cloned gene sequence of GPV YBLJ strain in GenBank （JN836326）, a pair of specific primers with Hind and Xho I sites were designed. VP3 gene was amplified by PCR, and subcloned into pcDNA 3.1. The recombinant plasmid was transferred into Vero cells with Lipofectamine 2000, and its expression was detected by RT-PCR and indirect immunofluorescent assay （IFA）. The results showed that the eukaryotic expression vector of pcDNA-VP3 was successfully constructed, a 1 605 bp specific band appeared in the experimental group by RT-PCR, the expression of VP3 was detected by IFA, which laid a foundation for development of GPV nucleic acid vaccine.

关 键 词：鹅细小病毒 VP3基因 真核表达 

分 类 号：S852.659.2[农业科学—基础兽医学]