表达鹅细小病毒VP3基因重组腺病毒的构建与鉴定

Construction and Identification of Recombinant Adenovirus Expressing VP3 Gene of Goose Parvovirus

[目的]构建表达鹅细小病毒(GPV)VP3基因的重组腺病毒,为机体免疫试验和免疫效果评价奠定基础。[方法]以重组质粒pc DNA-VP3为模板,以GPV-VP3为目的基因,构建GPV-VP3重组腺病毒载体,通过转染获得能稳定表达GPV-VP3基因的重组腺病毒,通过IFA和Western-blot检测GPV-VP3基因的表达情况。[结果]扩增到的GPV-VP3基因全长为1 605 bp,线性化的重组腺病毒穿梭质粒p CR259-VP3能在QBI-HEK293细胞中瞬时表达GPV-VP3基因,表达蛋白的分子量约为60 Ku。[结论]该重组腺病毒的构建将为GPV新型疫苗研发和后期体内试验奠定基础。

[Objective] The research aimed to construct recombinant adenovirus expressing VP3 gene of goose parvovirus and lay the foundation for making the immunity test and evaluating the immune effects.[Method] Taking recombinant plasmid pcDNA-VP3 as the template,using GPV-VP3 as target gene,the recombinant adenovirus vector of GPV-VP3 was constructed.The recombinant adenovirus that could stably express VP3 gene of goose parvovirus was obtained by transfection.The expression situations of GPV-VP3 were detected by IFA and Western-blot detection.[Result] The complete sequence of amplified GPV-VP3 gene was 1 605 bp.pCR259-VP3 plasmid could transiently express in QBIHEK293 cells.And the molecular weight of recombinant protein was about 60 Ku.[Conclusion] The construction of recombinant adenovirus laid a foundation for the research and development of a new vaccine of GPV and its experiment in vivo.