LAPTM4B启动子荧光素酶报告基因重组体的构建  

Construction of LAPTM4B promoter-luciferase reporter recombinants

作　　者：彭方毅[1] 张青云[2] 周柔丽[3] 邵根泽[3] 涂植光[1] 

机构地区：[1]重庆医科大学医学检验系,重庆400016 [2]北京大学临床肿瘤学院 [3]北京大学基础医学院细胞生物学与遗传学系,北京100083

出　　处：《重庆医科大学学报》2005年第3期337-339,共3页Journal of Chongqing Medical University

基　　金：北京市自然科学基金重点项目(编号:7041003)

摘　　要：目的:LAPTM4B基因是肝癌中高度表达的新基因。为下一步LAPTM4B启动子的活性分性,本文构建了LAPTM4B基因启动子调控的报告基因重组表达质粒。方法:以LAPTM4B基因组cDNA为模板扩增翻译起始位点上游DNA序列。PCR产物定向克隆到含荧光素酶报告基因的载体pGL3-Basic中,并经限制性内切酶消化鉴定。结果:通过酶切鉴定和基因测序,证明成功地构建了三种不同长度的pGL3-LAPTM4B-Luc荧光素酶表达质粒,形成了系列的LAPTM4B启动子重组体。结论:充分利用生物信息学资源,利用已知基因的启动子序列,可快速,准确地克隆已知基因启动子序列并构建启动子载体。LAPTM4B启动子荧光素酶报告基因重组体的构建为LAPTM4B启动子的活性分析奠定了基础。Objective:LAPTM4B is a novel gene which is over expressed in HCC.To evaluate LAPTM4B gene promoter on gene transcription,we construct luciferase reporter plasmid pGL3-LAPTM4B-Luc.Methods:The LAPTM4B cDNA was used as a template in the polymerase chain reaction to amplify its upstream regions from the translation initiation codon.The PCR product was directly cloned into the luciferase reporter vector pGL3-Basic.Results:Restriction enzymes digestion and nucleotide sequencing confirmed that the recombinant plasmids were correct without base mutation and deletion.Conclusion:A known gene promoter sequence could be freely obtained from Genbank database.This is very useful for the gene promoter cloning and promoter vector constructing.The construction of LAPTM4B recombinant plasmid lay a foundation of the analysis of promoter activities.

关 键 词：LAPTM4B 基因表达 启动子 

分 类 号：R735.7[医药卫生—肿瘤]