摘要
家蚕ser-1基因是中部丝腺中特异表达组织特异性基因。为研究家蚕ser-1基因在时空上的调控机制,用PCR的方法克隆了家蚕ser-1基因启动子并进行序列分析,进一步构建了由ser-1基因启动子驱动报告基因DsRed的新型表达质粒pSK-ser-DsRed-PolyA,并通过蚕体和家蚕BmN细胞进行了瞬时表达。结果显示,家蚕ser-1基因启动子的TATA框的保守序列为TATAAAA,位于-24～-30处,CAAT框位于-112～-115处;ser-1基因启动子可以驱动红色荧光基因DsRed在家蚕幼虫的中部丝腺组织和培养的BmN细胞中瞬时表达。
The gene encoding sericin 1 （ser 1） of silkworm （Bombyx mori） was specifically expressed in the middle silk gland. In order to understand the attractive model of the molecular mechanism governing spatially and temporally programmed transcription, the promoter of sericin-1 gene from Bombyx mori was cloned and sequenced. We constructed a new expression vector named pSK-ser-DsRed-PolyA in which the reporter gene DsRed was driven by sericin-promoter and characterized the promoter＇s activity by transient transfection expression assays in BmN cells and middle silk gland of Bombyx mori. As a result, it was showed that the sequence of TATA box was TATAAAA at position -24--30, the CAAT box at position -112--115, and the promoter of ser-1 could drive the red fluorescent gene, the transiently expression products could be observed both in the cultural Bran cells and the middle silk gland of Bombyx rnori.
出处
《科技通报》
2007年第6期828-834,共7页
Bulletin of Science and Technology
基金
国家重点基础研究发展计划"973"项目(编号2005CB121000)
国家自然科学基金项目(编号30571404)
江苏省研究生培养创新工程项目(编号ZY320506)
关键词
家蚕
丝胶蛋白基因
启动子
克隆
表达载体
Bombyx mori
sericin 1 gene
promoter
cloning
expression vector
