| 表达鹅细小病毒VP3基因重组腺病毒的构建与鉴定 |
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| 引用本文： | 陈海迪,高旭,张颖,许应天,张立媛,于清洋,鲁承.表达鹅细小病毒VP3基因重组腺病毒的构建与鉴定[J].安徽农业科学,2014(33):11751-11754. |
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| 作者姓名： | 陈海迪 高旭 张颖 许应天 张立媛 于清洋 鲁承 |
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| 作者单位： | 延边大学农学院动物医学系,吉林延吉,133002 |
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| 基金项目： | 吉林省自然科学基金项目 |
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| 摘 要： | 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pcDNA-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pCR259-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60Ku.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.
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| 关 键 词： | 鹅细小病毒 VP3基因 重组腺病毒 真核表达 |
Construction and Identification of Recombinant Adenovirus Expressing VP3 Gene of Goose Parvovirus |
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| Institution: | CHEN Hai-di,GAO Xu,ZHANG Ying(Department of Veterinary Medicine, Agricultural College of Yanbian University, Yanji, Jilin 133002) |
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| Abstract: | 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. |
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| Keywords: | Goose parvovirus VP3 gene Recombinant adenovirus Eukaryotic expression |
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