家蚕核多角体病毒P10基因启动子结构与功能分析  

STRUCTURE AND FUNCTION ANALYSIS OF THE PROMOTER OF BOMBYX MORI NUCLEARPOLYHEDROSIS VIRUS P10 CENE

作　　者：张燕[1] 张颖[1] 张志芳[1] 吴祥甫[1] 

机构地区：[1]中国科学院上海生物化学研究所

出　　处：《病毒学报》1995年第3期255-261,共7页Chinese Journal of Virology

摘　　要：前文曾报导过家蚕核多角体病毒（ＢｍＮＰＶ）Ｐ１０结构基因和启动子全顺序￣［１］。经突变法消除起始码ＡＴＧ，并分离上游－２３０位－＋１位含完整启动子片段，在此Ｐ１０启动子片段后拼接报导基因（昆虫荧光素酶基因，Ｌｕｃ，Ｌｕｃｉｆｅｒａｓｅ）构建成表达质粒ｐＢｍｐ１０－Ｌｕｃ。质粒ｐＢｍｐ１０－ＬｕｃＤＮＡ转染有野生型ＢｍＮＰＶ感染的家蚕Ｂｍ－Ｎ细胞可测出Ｌｕｃ基因的瞬时表达。由启动子５’端（－２３０位）向Ｌｕｃ基因节律缺失删节，获得缺失至－１３５、－１１１、－３８位的各质粒（ｐＢｍｐ１０－１３５Ｌｕｃ、ｐＢｍｐ１０－１１１Ｌｕｃ、ｐＢｍｐ１０－３８Ｌｕｃ）。功能检测证实除ｐＢｍｐ１０－３８Ｌｕｃ外，其它质粒都有Ｌｕｃ基因瞬时表达，说明－３８－－１１１位结构顺序是Ｐ１０启动子功能所必须的。在苜蓿尺蠖核多角体病毒（ＡｃＭＮＰＶ）感染的ＢｍＮＰＶ非敏感细胞Ｓｆ－９中也得到相同的结果，说明ＢｍＮＰＶＰ１０启动子和ＡｃＭＮＰＶＰ１０启动子的调节因子可能是相似的。e have reported the whole sequence of BmNPV (Bombyx mori nuclearpolyhedrosis virus)P10 structure gene and its promoter. The start codon ATG was deleted by point mutation and the fragment from-230bp upstream of ATG to +1nt,which contains the complete P10promoter, was separated, Luciferase gene which acts as reporter gene was fused with the promoter fragment to construct the plasmid pBmp10-Luc. The transient expression of Luc gene can bedetected when pBmp10-Luc was transfected to Bm-N cells which was already infected by wild type BmNPV. The plasmid pBmp10-Luc was deleted to -135bp、-111bp、-38bp bydeletion from -230bp of the promoter to Luc gene to construct the plasmid pBmp10-135Luc、pBmp10-111Luc、pBmp10-38Luc resepectively. Function analysis shows that all the plasmids have transient expression of Luc except pBmp10-38Luc,which indicatesthat sequence from -38bp-111bp is necessary for the function of p10 promoter. The sameresult can be obtained in AcMNPV-infected sf-9 cells which is not sensitive to BmNPV. This indicates that the regulatory factors for P10 promoter of BmNPV and AcMNPV are similar.

关 键 词：家蚕 核多角体病毒 P10基因 启动子 功能 

分 类 号：S884.5[农业科学—特种经济动物饲养]