BmNPV-ZJ iap1基因的克隆、序列分析及其对哺乳动物细胞NF-κB的调节作用研究(英文)  

Cloning and Sequence Analysis of iap1 Gene of Bombyx mori Nuclear Polyhedrovirus ZJ Strain and its Function on Regulating NF-κB Activity in Mammalian Cells

作　　者：吴金美[1] Robert WLim 吴祥甫[3] Grace Y Sun 吕鸿声[4] 

机构地区：[1]华东船舶工业学院生物技术学院,镇江212018 [2]University of Missouri-Columbia [3]中国科学院生物化学与细胞生物学研究所 [4]中国农业科学院蚕业研究所

出　　处：《蚕业科学》2003年第4期359-364,共6页ACTA SERICOLOGICA SINICA

基　　金：ThisworkwassupportedbythegrantsfromNationalNaturalScienceFounda tionofChina (No .3 0 0 70 5 79)andAG183 5 7fromNationalInstituteofHealth(NIH) ,USA

摘　　要：利用PCR法克隆得到家蚕核型多角体病毒镇江株 (BmNPV ZJ)的iap 1基因。序列同源分析表明 :BmNPV ZJ的iap 1全长为 85 8bp ,与BmNPV T3株的碱基同源性为 96 % ,与BmNPV T3株相比少了一段编码 7个氨基酸的区域 ,该缺失区域的两侧有着独特的结构 :缺失区的 5′侧为连续编码 7个天冬氨酸的序列 ;缺失区的 3′侧为连续编码3个天冬氨酸的序列。NCBI的DART(DomainArchitectureRetrievalTool)功能域搜索表明 ;BmNPV ZJ的iap1含有 2个杆状病毒BIR功能域 ,但不含RING区域。BmNPV的iap是否具有抗凋亡作用迄今尚未见报道。利用以NF κB为探针的凝胶阻滞分析表明 ,BmNPV ZJ的iap1转染鼠的pc12细胞 ,可逆转肿瘤坏死因子TNFα处理pc12细胞引起的核因子 κB(NF κB)的激活。BmNPV ZJ的iap1对NF κB的作用途径及作用方式 。The iap1 gene of Bombyx mori nucleopolyhedrovirus Zhenjiang strain was amplified by Polymerase Chain Reaction and cloned into pGEM-Teasy vector and sequenced. Sequence analysis results showed that iap1 of BmNPV-ZJ contains 858 base pairs of nucleotides, with 96% identity in nucleotides, but missing a region which encodes seven continuous aspartic acids, compared to that of BmNPV-T3 strain. DART search result (Domain Architecture Retrieval Tool) indicated that the iap1 of BmNPV-ZJ contained two BIR (baculovirus IAP-like repeat) domains, but lacked RING domain. It is not known yet if the iap of BmNPV has any function on apoptosis. Electrophoretic Mobility Shift Assay using NF-κ B as a probe has shown that transfection of BmNPV-ZJ iap1 gene into mouse pc12 cells could reverse the effect of TNF-α on NF-κ B binding activity. The action and its pathway of BmNPV-ZJ iap 1 on NF-κ B in pc12 cells and other mammalian cells are under way.

关 键 词：BmNPV—ZJiap基因 克隆 序列分析 哺乳动物细胞 NF-ΚB 调节作用 PCR 家蚕 核型多角体病毒镇江株 碱基同源性 凝胶阻滞分析 pcl2细胞 凋亡 

分 类 号：Q963[生物学—昆虫学]