小鼠BCMA基因启动子活性研究

Promoter activity of B-cell maturation antigen gene in mice

目的:探索B淋巴细胞成熟抗原(BCMA)基因5′上游序列的启动子活性,为进一步研究BCMA基因的表达调控机制提供实验依据。方法:构建由BCMA基因5′上游-820^+145、-607^+145、-359^+145、-157^+145、-93^+145 5个片段驱动的荧光素酶报告载体pGL3-B820、pGL3-B607、pGL3-B359、pGL3-B157、pGL3-B93,通过转染J558L、293T、HeLa细胞检测荧光素酶的表达,观察荧光素酶相对活性。结果:5种5′删除体的转录激活性由大到小为pGL3-B157>pGL3-B607>pGL3-B359>pGL3-B93>pGL3-B820。pGL3-B157在3种细胞中的转录激活能力为J558L>HeLa>293T,且在J558L中的转录激活能力明显强于Hela和293T细胞。结论:BCMA基因5′上游序列"-820bp^+145bp"具有启动子活性,核心启动子可能位于-93^+145区域中。

AIM： To investigate the promoter activity of BCMA gene 5′-flanking regionfor the study the regulation mechanism of BCMA expression.METHODS： Luciferase reporter plasmids containing 5′-flanking region of BCMA gene and serial deletions of the fragment were constructed.The luciferase expression was observed after these reporters were transfected into J558L,293T and Hela cells.RESULTS： The highest transcriptional activation was expressed in pGL3-B157 reporter,followed by pGL3-B607,pGL3-B359,pGL3-B93,and pGL3-B820 in sequence.The highest transcriptional activation was in J558L cell.CONCLUSION： The 5′-flanking sequence from-820 to ＋145 bp of BCMA is of promoter activity.Serial deletion analysis of the promoter region of BCMA gene suggests the sequence from-93 to ＋145 could be a core promoter region.