杆状病毒Splt MNPV SL136蛋白的功能  

Functional Analysis of SpltMNPV Envelope Protein SL136

作　　者：张萍[1] 庞义[1] 杨波[1] 李朝飞[1] 尹崇[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》2002年第3期333-337,共5页

基　　金：国家自然科学基金 (No .39730 0 30和No .3980 0 0 92 );国家重大基础研究项目 ( 973) (No .G2 0 0 0 0 16 2 0 9)资助~~

摘　　要：先前的研究发现斜纹夜蛾核多角体病毒 (Spodopteralituramulticapsidnucleopolyhedrovirus,SpltMNPV)基因组中Sl136表达产物具有膜融合功能。通过RT PCR检测了该基因的转录时相 ;制备该蛋白质的多克隆抗血清 ,SDS PAGE、Western印迹实验证明SL136蛋白质是芽生型病毒粒子特有的蛋白质。该基因在斜纹夜蛾离体细胞系中表达产物分子量为 86、6 5kD的两条带 ,后者与芽生型病毒粒子中检测到的一条蛋白质带分子量基本一致。另外 ,细胞酶联免疫吸附测定 (cellenzyme linkedimmunosorbantassay ,CELISA)实验证明SL136蛋白可分布于重组病毒Bac Sl136和野生型SpltMNPV分别感染的Hi5、Sl zsu 1细胞表面 ,并进行了定量分析。生物测定结果表明 ,弗林蛋白酶 (furin)抑制剂对于病毒感染力没有明显的影响 。Our previous research showed that the Spodoptera litura multinucleocapsid nucleopolyhedrovirus (SpltMNPV) Sl136 product had a function to be fused with membrane when expressed alone. In this study, the Sl136 and its product were characterized. RT-PCR results showed that Sl136 could be transcribed at 6 h post infection, indicating it was an early gene. Then the antiserum against SL136 protein was generated and utilized to verify that SL136 was a BV envelope protein, by using SDS-PAGE and Western blot. Two main protein bands in SpltMNPV-infected Sl-zsu-1 cells were detected; their molecular weights were about 86 kD and 65 kD, respectively. It was found that the size of smaller band coincided with a major band of BV envelope proteins. SL136 protein was transferred to cell surfaces both in SpltMNPV-infected Sl-zsu-1 cells and recombinant Bac-Sl136-infected Hi5 cells, as detected by cell ELISA(cell enzyme-linked immunosorbant assay, CELISA). From the bioassay results, it was found that furin cleavage of SL136 was not necessary for viral propagation, and inhibition of its glycosylation decreased the BV virulence.

关 键 词：杆状病毒 SPLTMNPV SL136蛋白 功能 斜纹夜蛾核多角体病毒 芽生型病毒 囊膜蛋白 

分 类 号：Q51[生物学—生物化学]