论文部分内容阅读
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858 bp,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(Domain Architecture Retrieval Tool)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-κ B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TNF α处理pc12细胞引起的核因子-κ B(NF-κ B)的激活.BmNPV-ZJ的iap1对NF-κ B的作用途径及作用方式,以及其对哺乳动物细胞的细胞凋亡相关作用的研究正在进行之中.“,”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.