斜纹夜蛾核型多角体病毒Ⅱ型(SpltMNPV Ⅱ)ORF117基因的结构和功能研究  

Structure and Function of ORF117 of Spodoptera litura multicapsid nucleopolyhedrovirus Ⅱ

作　　者：刘兴健[1] 江峰[1] 王石宝[1] 李轶女[1] 张志芳[1] 胡小元[1] 

机构地区：[1]中国农业科学院生物技术研究所,北京100081

出　　处：《中国农业科技导报》2016年第1期66-72,80,共8页Journal of Agricultural Science and Technology

基　　金：国家863计划项目(2011AA100603);国家973计划项目(2012CB114600)资助

摘　　要：为了研究斜纹夜蛾核型多角体病毒Ⅱ型(Splt MNPVⅡ)中ORF117基因(编码GP117蛋白)的结构和功能,根据其核苷酸序列和氨基酸序列进行了生物信息学分析,以此为基础对该基因的启动子活性和转录时相进行分析,表达纯化了GP117蛋白的部分序列以制备豚鼠来源多克隆抗体,并利用该抗体对ORF117基因的表达时相和GP117蛋白在细胞中的定位进行了分析。生物信息学分析表明该基因全长1 203 bp,编码400个氨基酸的蛋白质,预测分子量为45.5 k Da,等电点为5.06;启动子活性和转录时相分析结果表明ORF117是一个晚期表达的基因;原核表达抗原免疫制备的多克隆抗体效价可以达到1∶3 200,利用该抗体对其表达时相的分析进一步验证了ORF117为晚期表达基因,免疫荧光检测表明编码的蛋白多存在于细胞质中。The structure and function of Spodoptera litura muhicapsid nucleopolyhedrovirus Ⅱ （SphMNPV Ⅱ ） ORF117 gene were studied. The gene and protein sequences of ORF117 （coden protein GP117） were analyzed by bioinformatics methods. The promoter activities and the time course of mRNA transcription were analyzed. Part of GP117 protein was expressed and purified to preparing the polyclonal antibody. The expression phase of ORF1l7 and cellular location of GP117 were analyzed using the antibody. Sequence analysis demonstrated that this gene has a 1 203 bp ORF, encoding 400 amino acids with a predicted molecular weight of 45.5 kDa and a predicted isoelectric point of 5.06. Both promoter activities analysis and the time course of mRNA transcription showed that ORF117 was a late gene. Western blotting was used to detected the gene ORF117＇s expression phase and the result identified with the transcription phase. The result of cellular localization of ORF117 showed that the GP117 existed in cytoplasm more than nucleus.

关 键 词：斜纹夜蛾 核型多角体病毒Ⅱ型 ORF117 转录时相分析 表达时相分析 

分 类 号：Q78[生物学—分子生物学]