杆状病毒Splt MNPV Sl136基因的克隆、表达及其产物功能  

Cloning and Expression of SpltMNPV Sl136 Gene and Functions of the Expressed Product

作　　者：张萍[1] 杨波[1] 庞义[1] 苏德明[2] 

机构地区：[1]中山大学生物防治国家重点实验室,广州510275 [2]复旦大学生命科学学院病毒学研究室,上海200433

出　　处：《生物化学与生物物理学报》2001年第5期531-536,共6页

基　　金：国家自然科学基金 (No .39730 0 30 ;No .3980 0 0 92 )资助项目&&

摘　　要：计算机分析斜纹夜蛾核多角体病毒 (SpltMNPV)基因组序列 ,发现第 136个读码框基因表达产物具有病毒囊膜蛋白的基本特征。计算机预测的SL136蛋白氨基酸序列N端具有信号肽 ,C端具有跨膜区 ,N端区还有一个许多病毒融合蛋白共有的卷曲螺旋结构。通过PCR扩增 ,我们克隆了Sl136基因并分别构建了原核表达载体pBVSl136及重组病毒表达载体。SDS PAGE结果表明Sl136基因在大肠杆菌和昆虫细胞中均获得了较高的表达。另外 ,我们还构建了一个瞬时表达载体pUCSl136 ,单独转染Sl zsu 1细胞后 ,低 pH值环境可诱导细胞发生膜融合并形成合胞体。这些实验结果表明 。By computer assisted analysis, it was revealed that ORF136 gene product in SpltMNPV genome had the basic properties of membrane protein. A putative signal peptide was present at the N terminal and a transmembrane region near the C terminal of SL136 protein. In the N terminal half region, there was a coiled coil domain, which is a typical feature of a number of viral fusion proteins. After PCR amplification, a recombinant plasmid pBVSl136 and a recombinant AcMNPV containing Sl136 were constructed, in order to express Sl136 gene in E.coli and insect Hi5 cells, respectively. The SDS PAGE results showed that both expression levels were high. Cell membrane fusion was induced in the Slzsu1 cells, which had been transfected with Sl136 gene alone, by lowering pH of the medium. These results suggested that SL136 protein may be an envelope fusion protein.

关 键 词：斜纹夜蛾 杆状病毒 融合蛋白 表达产物 SPLTMNPV Sl136基因 

分 类 号：Q785[生物学—分子生物学]