Construction of LAPTM4B promoter-luciferase reporter recombinants

PENG Fangyi; et al

doi:10.13406/j.cnki.cyxb.2005.03.008

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Construction of LAPTM4B promoter-luciferase reporter recombinants

更新时间：2024-01-17

- Construction of LAPTM4B promoter-luciferase reporter recombinants
- Journal of Chongqing Medical University Issue 3, Pages: 337-339(2005)
- 作者机构：
  
  1. 重庆医科大学医学检验系
  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
  4. 重庆医科大学医学检验系,重庆,400016
  5. 北京肿瘤医院检验科
  6. 北京市肿瘤防治研究所免疫研究室
  7. ,北京,100083
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2005.03.008
  CLC： R735.7
- Published：2005，
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[1]彭方毅,张青云,周柔丽,邵根泽,涂植光.LAPTM4B启动子荧光素酶报告基因重组体的构建[J].重庆医科大学学报,2005(03):337-339.

PENG Fangyi, et al. Construction of LAPTM4B promoter-luciferase reporter recombinants. [J]. Journal of Chongqing Medical University (3):337-339(2005)
[1]彭方毅,张青云,周柔丽,邵根泽,涂植光.LAPTM4B启动子荧光素酶报告基因重组体的构建[J].重庆医科大学学报,2005(03):337-339. DOI： 10.13406/j.cnki.cyxb.2005.03.008.

PENG Fangyi, et al. Construction of LAPTM4B promoter-luciferase reporter recombinants. [J]. Journal of Chongqing Medical University (3):337-339(2005) DOI： 10.13406/j.cnki.cyxb.2005.03.008.

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。

Abstract

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

Keywords

LAPTM4B (lysosome associate protein transmembrane 4 beta)gene expressionpromoter

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