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

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.