人BAFF-R基因启动子活性的初步研究

Activity of human BAFF-R gene promoter

目的以B淋巴细胞刺激因子受体BAFF-R基因5'上游区序列为研究对象,初步鉴定、分析该基因的启动子所在区域。方法克隆BAFF-R基因5'侧翼区1823 bp序列,构建8个含有不同长度启动子序列的荧光素酶报告基因表达质粒pGL3-B1～B8,将这8个序列缺失重组质粒与psv-β-gal质粒共转染细胞,检测荧光素酶的相对活性,确定启动子所在区域。结果8个重组质粒中pGL3-B2、pGL3-B3、pGL3-B5、pGL3-B6启动子的活性较低;pGL3-B7启动子的活性最强,pGL3-B8启动子活性最低。结论BAFF-R基因5'端-288～-430、-712～-868和-1420～-1562三个区段可能存在转录沉默子元件,-617～-712和-1277～-1420区段存在转录增强子元件,BAFF-R基因的核心启动子可能位于-1420～+261区域。

We would identify and analyze the promoter of BAFF-R gene 5＇-flanking region,so as to define the regulation mechanism of BAFF-R expression.We cloned 5＇-flanking region of BAFF-R and constructed luciferase reporter plasmids containing the 5＇-flanking region of BAFF-R gene and serial deletions of the fragment.The all expression vectors were co-transfected with psv-β-gal vector into cells.The relative promoter activity of pGL3-B1,pGL3-B4,and pGL3-B7 were higher,while pGL3-B7 was the highest,as compared with others;the relative promoter activity of pGL3-B8 was the lowest in these plasmas.The results mean that transcription silence elements may exist in the region -288--430,the region -712--868,as well as the region -1420--1562.While transcription enhancer elements may exist in the region -617--712 and the region -1277--1420.Serial deletion analysis of the promoter region of BAFF-R gene suggests the sequence -1420-＋261 may be a core promoter region.