家蚕核多角体病毒解旋酶基因启动子功能区域缺失分析  

Identification of Functional Region of Helicase Gene Promoter in Bombyx mori Nuclear Polyhedrosis Virus

作　　者：肖庆利[1] 张志芳[1] 易咏竹[1] 何家禄[1] 吴祥甫[2] 

机构地区：[1]中国农业科学院蚕业研究所,农业部家蚕生物技术重点开放实验室,镇江212018 [2]中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031

出　　处：《生物化学与生物物理学报》2002年第5期560-564,共5页

基　　金：国家自然科学基金资助项目 (No .3 9970 5 71)

摘　　要：杆状病毒DNA解旋酶是病毒复制所必需的。瞬时表达分析显示 ,家蚕核多角体病毒解旋酶基因启动子属于延迟早期基因启动子。通过PCR技术在该启动子区产生的一系列缺失分析表明 ,解旋酶基因启动子的基础转录调控区主要位于ATG上游 - 5 1 0～ - 4 1 0bp之间。当只保留ATG上游 98bp区段时 ,仍可测到该启动子的基础活性。在病毒因子存在下 ,将启动子区域删除到ATG上游 - 4 1 0bp时 ,对启动子活性影响不大 ;若继续删除 ,则其活性显著下降。据此推测对病毒因子响应的启动子区段应主要位于ATG上游 - 4 1 0～ - 30DNA helicases are essential for replication of baculoviruses. It was found that the helicase gene promoter of Bombyx mori nuclear polyhedrosis virus, including 510 bp upstream of ATG, had both early and late RNA initiation sites and could be recognized by cellular RNA polymerase. Transient expression assays in uninfected Sf 21 cells indicated that the helicase gene promoter could be classified as a delayed early gene promoter. Deletion analysis by PCR showed that the regulation region of its basic transcription was mainly within -510 to -410 bp upstream of ATG. However, the basic activity was still detected with a deletion to -98 bp relative to ATG. In the presence of viral factors, deletion between -510 to -410 bp relative to ATG did not significantly reduce the promoter activity compared to the full length promoter (510 bp). The remarkable reduction in the promoter activity was observed with continuous deletions. It suggests, therefore, that cis acting elements responsive to viral factors are mainly located within the range of -410 to -309 bp upstream of ATG.

关 键 词：瞬时表达 家蚕核多角体病毒 解旋酶基因 启动子 功能区域 缺失分析 

分 类 号：S884.5[农业科学—特种经济动物饲养]