Expression of SpltMNPV p49 Gene in Escherichia coli and Determination of The Time Course of Its Expression in Insect Cells
斜纹夜蛾核多角体病毒p49基因在大肠杆菌中的表达及其在昆虫细胞中的表达时相分析

The apoptotic suppressor p49 gene was amplified by PCR from the genome of Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) and cloned into the pMD18-T vector. Sequence analysis verified the cloned fragment. The gene was then inserted into thioredoxin fusion expression vector pThioHis C and expressed stably in E.coli BL21(DE3) by induction with IPTG. The expression products were about 30% of the total bacteria proteins and exsisted as inclusion bodies. Specific antibodies with a high titre of over 1:10000 were obtained by immunizing rabbits with the purified fusion protein and was used to determine the time course of P49 expression in SpltMNPV-infected Sl cells. The results showed that P49 expression startups within 3 hours post infection and remains low level during the whole course of infection.