人类mtDNA D-Loop区位点变异对基因表达的影响  

The Effect of Gene Expression of Loci Mutation of Human Mitochondrial DNA D-Loop Region

作　　者：龚丽景[1] 胡扬[1] 李燕春[1] 梅涛[1] 

机构地区：[1]北京体育大学,北京100084

出　　处：《体育科学》2014年第5期3-8,共6页China Sport Science

基　　金：国家自然科学基金项目(31071032)

摘　　要：Case-Control研究表明,人线粒体基因(mtDNA)D-Loop区mt235、mt514-515、mt16183、mt16290和mt16319多态位点与有氧运动能力有关。拟从下游基因表达做进一步的研究,以探讨上述多态位点的分子调控机理。方法:采用全基因组合成法合成包含上述多态位点的DNA序列,分别连接到pGL3-promoter载体构建重组质粒,将重组质粒转染至293T细胞,观察携带5对mtDNA质粒对下游双荧光素酶报告基因的表达及荧光素酶mRNA表达的影响。结果:携带mt235A、mt514-515CA、mt16183A和mt16319G的载体转染细胞的双荧光素酶基因表达量显著高于mt235G、mt514-515Del、mt16183C和mt16319A(P<0.01);携带mt16290两种基因型的质粒转染细胞报告基因表达没有明显差异。与pGL3-promoter载体相比,携带mt16319A的质粒转染细胞荧光素酶mRNA极显著下降(P<0.01),携带mt16290位点的两种质粒对报告基因mRNA的影响没有差异。结论:mt235、mt514-515、mt16183和mt16319多态位点影响荧光素酶基因表达,mt235A、mt514-515CA和mt16183A上调基因转录和翻译,mt16319A下调基因转录。这些基因表达上的差异可能是影响人有氧运动能力的重要调控因素。Objective ： The gene expression differences between variants of mt 235, mt 514- 515, mt16183 ,mt16290 and mt16319 correlated with enduance performance in human mtDNA D- Loop region were studied. Methods：The gene sequences contained above loci were synthesized respectively,and then constructed with pGL3-promoter vector. The vectors were transfected into 293T cells, and the expression activity and mRNA of the reporter gene in the recombinant expression vectors with different alleles were detected by a dual-luciferase reporter assay system and RT-PCR. Results： Higher relative luciferase activities were detected significantly in vectors containing mt235A, mtS14-515CA, mt16183A and mt16319G than mt235G, mt514- 515Del, mt16183C and mt16319A（P^0.01）. There was no effect for reporter gene expres- sion in vectors containing mt 16290 locus. Compared with pGL3-promoter, luciferase gene mRNA level of transfected pGL3-16319 A cell was decreased significantly, but no differences between vectors contained mt16290. Conclusion： The variation of mt235A, mt514-515CA and ml 16183 A can up-regulate the gene transcription and expression in this study, maybe affecting the gene function, mt16319A can down-regulate the gene transcription. The gene expression differences maybe are the important regulated factors.

关 键 词：基因多态性 线粒体基因 基因表达 D-LOOP区 质粒转染 

分 类 号：G804.7[文化科学—运动人体科学]