-514 bp及-250 bp位点基因多态性对肝脂酶转录活性的影响  

The effect of Hepatic Lipase Gene Promoter Polymorphisms on the Transcriptional Activity

作　　者：蔡泽园[1] 赵莉莉[1] 张蕊[1] 毛用敏[1] 崔让庄[1] 

机构地区：[1]天津市胸科医院心血管病研究所,300051

出　　处：《天津医药》2012年第8期803-805,I0002,共4页Tianjin Medical Journal

摘　　要：目的:探讨同时含肝脂酶(HL)启动子-514bp和-250bp多态位点的野生型及变异纯合子型的荧光素酶表达质粒对HL转录活性的影响。方法:PCR法扩增含不同目的基因的DNA片段,将目的基因插入到pUCm-T质粒中,获取含SacI及BglⅡ酶切位点的目的基因,将该基因与含荧光素报告基因的pGL2质粒相连,构建pGL2-HL/-514T+-250A变异纯合型以及pGL2-HL/-514C+-250G野生型荧光素酶表达质粒,转染至HepG2细胞内表达,双荧光素酶检测系统检测不同重组质粒荧光素酶报告基因的活性。结果:(1)实验成功构建了同时含HL启动子部位-514/-250位点的野生型和变异纯合型的荧光素酶表达质粒。(2)pGL2-HL/-514T+-250A变异纯合型的荧光素酶相对表达活性明显低于野生型荧光素酶表达活性(P<0.01)。结论:HL启动子-514及-250位置基因的联合变异可直接影响HL的转录活性。Objective： To investigate the effect of recombinant plasmids containing the double luciferase report gene and the hepatic lipase （HL） promoter-514/-250 sites on HL transcriptional activity. Methods： DNA fragments containing different target genes were amplified by PCR. Target genes were inserted into the plasmid pUCm-T to obtain the target gene containing SacI and Bgl I] restriction sites. The [uciferase reporter vectors containing HL promoter-514T/-250A and -514C/-250G polymorphisms were constructed, and tested by Dual Luciferase Reporter Assay System. Results： The lu- ciferase reporter vector containing HL promoter -514T ＋ -250A presented higher HL transcriptional activity than that of vec- tor with -514C ＋ -250G（P 〈 0.01 ）. Conclusion： HL promoter -514C together with -250G alleles were associated with high- er HL transcriptional activity, which proved that HL promoter polymorphisms can influence the HL transcriptional activity in the molecular mechanism.

关 键 词：肝脂酶 启动区(遗传学) 多态现象 遗传 杂合子 纯合子 转录 遗传 基因表达 

分 类 号：R541.4[医药卫生—心血管疾病]