文献类型：期刊文献

中文题名：表达鹅细小病毒VP3基因的重组腺病毒的构建及其免疫原性分析

英文题名：Construction and immunogenicity analysis of recombinant adenovirus bearing goose parvovirus VP3 gene

作者：白翠玲[1];刘铀[2];陈绍红[2];赵云涛[2];刘艳芬[1]

机构：[1]广东海洋大学农学院,广东湛江524088;[2]广东海洋大学生化中心,广东湛江524088

年份：2014

卷号：44

期号：11

起止页码：1114

中文期刊名：中国兽医科学

外文期刊名：Chinese Veterinary Science

收录：CSTPCD、、北大核心2011、北大核心、CSCD、CSCD2013_2014

基金：广东省科技计划项目(2007A020200006-8)

语种：中文

中文关键词：鹅细小病毒;VP3基因;重组腺病毒;免疫原性

外文关键词：goose parvovirus;;VP3gene;;recombinant adenovirus;;immunogenicity

中文摘要：为构建表达鹅细小病毒(GPV)VP3基因的重组腺病毒,并检测其对雏鹅的免疫效果,将GPV VP3基因克隆至腺病毒表达系统穿梭质粒,构建了重组穿梭质粒pacAd-VP3;将经PacⅠ酶切线性化的重组穿梭质粒与骨架质粒共转染HEK293AD细胞,得到重组腺病毒rAd-VP3。采用RT-PCR、Western-blot和间接免疫荧光试验鉴定GPV VP3基因在真核细胞中的表达,用rAd-VP3免疫1月龄雏鹅,检测血清抗GPV蛋白抗体水平。结果显示,重组腺病毒rAd-VP3能感染HEK293AD和GEF细胞并表达GPV VP3蛋白,增殖至第6代重组病毒的效价可达108.23 TCID50/0.1mL;rAd-VP3能诱导雏鹅产生抗GPV特异抗体,免疫后第28天血清中抗GPV VP3蛋白抗体仍维持较高水平。结果表明,rAd-VP3具有良好的免疫原性,可作为一种安全有效的病毒活载体疫苗用于鹅细小病毒病的免疫预防。

外文摘要：To construct a recombinant adenovirus expressing VP3 gene of goose parvovirus(GPV)and to test immunological efficiency on the immunized goslings,the recombinant adenovirus,named as rAdVP3,was constructed through homologous recombination internally in the HEK293 AD cells after co-transfection of the PacⅠ-linearized backbone plasmid and the shuttle plasmid containing the GPV VP3 gene.The recombinant adenovirus was identified by RT-PCR,Western-blot and indirect immunofluorescence assay(IFA).Then 1-month-old goslings were immunized with rAd-VP3 and the serum antibody against GPV VP3 protein was detected.In result,the GPV VP3 protein could be efficiently expressed by the recombinant adenovirus rAd-VP3 in HEK293AD cells and GEF cells,and the virus titer of the 6th generation was up to 108.23 TCID50/0.1mL.The serum antibody against GPV could be detected on day 14 after inoculation of goslings with rAd-VP3,and retained at a high level for following 14 days.The result showed that rAd-VP3 had good immunogenicity,and could be used as a safe,effective vaccine for prevention of goose parvovirus infection.