TLR4基因3′未翻译区内11367位点突变对荧光素酶表达的影响  

Influence of 3′ untranslated region single nucleotide polymorphism of TLR4 gene on luciferase gene expression

作　　者：段朝霞[1] 朱佩芳[1] 王正国[1] 蒋建新[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所第四研究室

出　　处：《第三军医大学学报》2006年第21期2115-2118,共4页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划资助项目("973"项目)(G199054203);国家杰出青年科学基金资助项目(30325040)~~

摘　　要：目的分别构建含TLR4基因3′未翻译区及11 367位点突变的含荧光素酶基因的表达质粒,应用双荧光报告系统观察TLR4基因3′未翻译区对荧光素酶基因表达的影响。方法采用基因工程的方法构建含TLR4基因3′未翻译区及11 367位点突变的含荧光素酶基因的表达载体,用LipofectAMINETM2000转染HEK293细胞,化学发光法检测荧光素酶活性。结果经酶切及测序鉴定显示,TLR4基因3′未翻译区及11 367位点突变的片段正确插入荧光素酶表达载体pGL3-promoter中,应用双荧光报告系统检测显示pGL3-11 367G质粒转染后荧光素酶活性显著高于pGL3-11 367C。结论TLR4基因3′未翻译区单核苷酸多态性能抑制荧光素酶基因的表达。Objective To construct two luciferase gene expression vectors which contain TLR4 gene 3＇ untranslated region and 3＇ untranslated region 11 367 site-mutant respectively, and use double-fluorescence reporter system to observe the influence of 3＇ untranslated region single nucleotide polymorphism of TLR4 gene on luciferase gene expression. Methods The luciferase gene expression vectors which contain TLR4 gene 3＇ untranslated region and 3＇ untranslated region 11 367 site-mutant segment respectively were constructed by the gene engineering technology, and then transfected into the HEK293 cells by LipofectAMINETM2000, finally the activities of fluorescence were detected by the chemic-luminescence. Results The sequence of human TLR4 gene 3＇ untranslated region was identical with that of TLR4 gene 3＇ untranslated region recorded in the Gen- Bank. The analysis for recombinant plasmid DNA digested by Xba I and Sma I demonstrated that the TLR4 gene 3＇ untranslated region and 3＇ untranslated region 11 367 site-mutant segment were inserted exactly into pGL3-promotor respectively. The activities of fluorescence of pGL3-11 367G were higher than pGL3-11 367C （P 〈0.05）. Conclusion TLR4 gene 3＇ untranslated region single nucleotide polymorphism can inhibit the expression of luciferase gene.

关 键 词：TLR4 单核苷酸多态性 荧光素酶 定点突变 

分 类 号：Q786[生物学—分子生物学] Q782