载脂蛋白M基因启动子克隆及功能初步研究

Primary study on the function and cloning of the apoM gene promoter

目的克隆apoM基因的上游启动子序列,构建启动子不同截短片段的荧光素酶报告基因,并观察其不同截短片段的启动子活性,为进一步研究apoM基因的表达调控机制奠定基础。方法在对apoM基因生物信息分析的基础上,采用5’RACE确定了apoM基因的转录起始点,构建了6种不同长度apoM基因启动子序列的荧光素酶报告基因表达质粒pGL3-Basic(A-F),将它们瞬时转染HepG2细胞,并检测其荧光素酶活性。结果 pGL3-Basic-C的荧光素酶活性最高。结论 -401～-621bp可能是apoM的核心启动子区。

Objective To study the molecular mechanisms that regulate the expression of apoM gene, construct a series of firefly luciferase report gene basic vectors for promoter region of apoM gene. Methods After analysing the 5flanking region of apoM, determined the transcriptional start sites of apoM gene by 5 RACE. Then, constructed six luciferase expression vectors pGL3-Basic（A-F）. All expression vectors were transected into HepG2 cells and tested these promoter fragments transcrip- tional activities by luciferase assay. Results The pGL3-Basic-C showed the highest level of basal activi- ty. Conclusion The --401--621 may be the core promoter region of apoM gene.