家蚕CIb1基因启动子活性分析  

Activity of Clb1 Gene Promoter From Bombyx mori

作　　者：赵巧玲[1,2,3] 唐顺明[2,3] 易咏竹[2,3] 赵秀振[2] 沈兴家[2,3] 朱良均[1] 张国政[2,3] 郭锡杰[2,3] 

机构地区：[1]浙江大学动物科学学院 [2]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 [3]江苏科技大学生物与环境工程学院

出　　处：《蚕业科学》2006年第4期477-481,共5页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划"973"项目(编号2005CB121000);江苏省自然科学基金前期预研重大项目(编号BK2004206);江苏省高技术研究计划(编号BG2005302)。

摘　　要：用PCR方法从家蚕基因组DNA扩增了家蚕胰凝乳蛋白酶抑制剂b1基因(CIb1)4个不同长度的启动子片段,构建由其驱动的萤火虫荧光素酶基因报告质粒,在脂质体介导下转染家蚕BmN细胞,体外分析了该基因启动子的活性。结果表明,家蚕CIb1基因启动子在BmN细胞中有微弱的转录活性；野生型BmNPV感染能增强启动子活性；hr3增强的不同长度CIb1基因启动子片段的活性有显著差异,提示转录起始位点上游-74～-1nt包含了启动子的基本元件,而在-687～-465nt、-465～-317nt和-317～-74nt存在着主要的顺式元件。试验结果有助于阐明CIb1的表达调控机理和对家蚕天然性免疫的理解。Four heterogeneous length promoters of chymotrypsin inhibitor b1 gene （CIb1） were generated from the genomic DNA of silkworm, Bombyx moil, by PCR, Report plasmids containing a luciferase gene, driven by CIb1 gene promoters, were constructed to transfect BmN cells mediated by lipofectin. The activity of CIb1 gene promoter was analysed in vitro. The results demostrated that CIb1 gene promoter showed a week transcriptional activity in ovary-derived BmN cells; that infection of wild type Bombyx moil nucleopolyhedrovirus （BmNPV） enhanced the activity of CIb1 gene promoter, and that the transcriptional activity of hr3 enhanced CIb1 gene promoter differs from the length of nucleotide fragment, which imply that the nucleotide sequence from -74 to -1 nt upstream the transcription initiation site of CIb1 gene contains the basal transcriptional elements, and nucleotide sequence between -687 to -465 nt, -465 to -317 nt and -317 to -74 nt upstream the transcriptional initiation site harbors the main cis-acting elements.

关 键 词：家蚕 胰凝乳蛋白酶抑制剂 CIb1基因启动子 家蚕核型多角体病毒 同源重复区3 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q781[农业科学—畜牧兽医]