摘要
为构建表达禽流感病毒(AIV)血凝素(HA)基因的重组疱疹病毒(HVT),本研究将AIV A/Goose/Guangdong/3/96(H5N1)的HA基因克隆于真核表达质粒中,构建了转移载体pUAB-gpt-HA。通过同源重组及霉酚酸筛选技术,将HA基因表达盒插入到HVT US10基因区,构建了一株表达H5亚型AIV HA基因的重组病毒HVT(rHVT-US10-HA)。经PCR、间接免疫荧光、western blot及红细胞凝集试验鉴定,重组病毒能稳定表达具有生物学活性的HA蛋白。rHVT-US10-HA的构建为禽流感基因工程病毒活载体疫苗的研究奠定了基础。
In present study,the cDNA of HA gene from avian influenza virus（AIV） A/Goose/Guangdong/3/96（H5N1） was cloned into eukaryotic expressing vector to construct transfer vector pUAB-gpt-HA.The transfer vector was co-transfected with herpesvirus of turkey（HVT） total DNA into the chicken embryo fibroblast cells（CEF） to generate the recombinant virus by homologous recombinantion.The recombinant HVT（rHVT-US10-HA） carrying HA gene was obtained by gpt selection and identified by PCR.The expression of HA gene was verified by indirect immunofluorescence assay,western blot and hemagglutination test in infected CEF.This work provided a foundation for developing avian influanza vaccine based on live viral vector.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2010年第5期329-333,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
兽医生物技术国家重点实验室国家基本科研业务费课题(NKLVBP200803)
