斜纹夜蛾核型多角体病毒日本株(C3)ie0基因的序列分析和表达

Expression and Sequence Analysis of ie0 Gene from the Genome of Japanese strain(C3) in Spodoptera litura Multicapsid Nucleopolyhedrovirus(SpltMNPV)

从斜纹夜蛾核多角体病毒(Spodoptera litura multicapsid nucleopolyhedrovirus,SpltMNPV)日本株(C3)基因组中克隆了ie0基因。核苷酸序列分析表明,该克隆片断涵盖了864 bp的读码框及5′端启动子区475 bp和3′端终止区90 bp的序列。氨基酸序列分析显示,在SpltMNPV IE0蛋白N-端23～47位以及111～126位富含酸性氨基酸和疏水性氨基酸残基,C′端第225～271位具有典型的CX2CX14CCX4CX2CX15CX2C双锌指基序,该结构在所比较的9个昆虫杆状病毒IE0蛋白中是十分保守的。联配分析表明,SpltMNPV日本株(C3)的核苷酸序列和氨基酸序列与其他9种核多角体病毒的同源性有较大差异。以ie0基因为基础绘制的杆状病毒分子进化树将家蚕核多角体病毒(Bombyx mori Nu-cleopolyhedrovirus,BmNPV)与苜蓿银纹夜蛾核多角体病毒(Autographa californica multicapcid nucle-opolyhedrovirus,AcMNPV)归到同一分枝,这与以p10、gp37和egt基因为基础绘制的杆状病毒分子进化树结果一致。并进行了大肠杆菌表达和Western Blotting分析。

The ie0 gene was cloned from the genome of Spodoptera litura multicapsid nucleopolyhedrovirus （SpltMNPV） JP-C3 which was isolated in Japan,Nucleotide sequence analysis demonstrated that this clone included 864 bp open reading frame（ORF）, 475 bp of 5＇-end promoter region and 90 bp of 3＇-end terminator region. Amino acid sequence analysis showed that there were abundant acidic and hydrophobic amino acid residuals at the 23th to 47th sites and the 11 lth to 126th sites in N-terminus side of SpltMNPV IE0, a typical double zinc-finger structure at 225th to 271th sites in C-terminus side, which was conservative in IE0 of the nine comparative insect baculoviruses. On-line analysis indicated that nucleotide sequence and amino acid sequence of SphMNPV JP-C3 were distinct from those of the other NPVs. The phylogenetic tree of baculoviruses based on the ie0 gene incorporated BmNPV （Bombyx mori Nucleopolyhedrovirus） and AcMNPV（Autographa californica muhicapcid nucleopolyhedrovirus） into the same branch, which was identical to those based on p10, gp37 and egt genes, and expressed in E.coli BL21,identified by Western Blotting.