棉铃虫核型多角体病毒ORF33基因的原核表达和在宿主细胞中亚细胞定位(英文)

Bacterial expression and cellular localization of Helicoverpa armigera nucleopolyhedrovirus Orf33 in infected host cells

对棉铃虫Helicoverpa ar migera核型多角体病毒HearSNPV的ORF33基因(ha33)进行克隆和原核表达,hass在E.coli中表达不完全,表达产物的大小为17kDa,小于预测的分子量28.4kDa。用纯化的原核表达产物免疫家兔,制备了多克隆抗体,应用多克隆抗体检测了HearSNPV感染的宿主细胞(HzAMI)中ORF33基因的表达,表达产物的分子量为31kDa。并通过共聚焦荧光显微镜方法,用多克隆抗体检测编码的蛋白在宿主细胞(HzAM1)中的亚细胞定位,发现ha33编码的蛋白存在于宿主细胞的细胞质中,并持续到感染后期。

Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus （HearSNPV） is a selective, highly infectious pathogen to H. armigera and has been extensively used for the control of this pest. This study presents the bacterial expression and localization of ha33 in infected host cells. The ha33 protein expressed in Escherichia coli had protein size of 17kDa. Western blot analysis using polyclonal antibody showed that the product of ha33 in infected Helicoverpa zea cells （HzAM1） was a 31kDa protein, larger than the theoretically predicted size of 28.4kDa. The confocal laser scanning microscope observation demonstrated that ha33 gene product was localized in the cytoplasm of infected HzAM1 cells beginning at 6 h p.i. and remained throughout the infection.