Construction and identification of recombinant adenovirus vector containing human β-catenin

BI Yang; et al

您当前的位置：

首页 >

文章列表页 >

Construction and identification of recombinant adenovirus vector containing human β-catenin

更新时间：2024-01-17

- Construction and identification of recombinant adenovirus vector containing human β-catenin
- Journal of Chongqing Medical University Issue 6, Pages: 561-564(2007)
- 作者机构：
  
  1. 重庆医科大学基础医学院生物化学与分子生物学教研室
  2. 重庆医科大学基础医学研究所
  3. 重庆医科大学基础医学院生物化学与分子生物学教研室,重庆,400016
- 作者简介：
- 基金信息：
- DOI：
  CLC： Q78
- Published：2007，
扫描看全文
[1]毕杨,黄佳祎,吴明军,张琴,张任飞,赵迎泽,冯涛.β-catenin腺病毒载体的构建及鉴定[J].重庆医科大学学报,2007(06):561-564.

BI Yang, et al. Construction and identification of recombinant adenovirus vector containing human β-catenin. [J]. Journal of Chongqing Medical University (6):561-564(2007)
[1]毕杨,黄佳祎,吴明军,张琴,张任飞,赵迎泽,冯涛.β-catenin腺病毒载体的构建及鉴定[J].重庆医科大学学报,2007(06):561-564. DOI：

BI Yang, et al. Construction and identification of recombinant adenovirus vector containing human β-catenin. [J]. Journal of Chongqing Medical University (6):561-564(2007) DOI：

摘要

目的:构建人β-catenin基因的重组腺病毒载体

为进一步研究Wnt/Wingless信号通路在肿瘤发生发展中的作用机制奠定重要基础。方法:以含有β-catenin的pcmv-βNS-FC质粒为模板PCR获得目的基因

将目的基因定向克隆至腺病毒穿梭质粒构建阳性重组pAd track-β-catenin

并经PmeI线性化后在BJ5183细菌中与腺病毒骨架质粒pAd Easy-1同源重组获得β-catenin重组腺病毒载体

再将PacI线性化的腺病毒质粒转染HEK293细胞包装重组腺病毒。结果:PCR及酶切鉴定证实目的基因正确克隆至腺病毒质粒中

目的基因序列与GENEBANK报道一致

荧光显微镜可观察到GFP的表达。结论:成功构建pAd-β-catenin腺病毒质粒载体

并在HEK293细胞中包装出重组腺病毒。

Abstract

Objective:To construct the recombinant adenovirus vector containing humanβ-catenin gene by homogenous recombination in E.coli for later research of the mechanism of Wnt/Wingless signal pathway in tumorigenesis.Methods:The target gene was amplified from pcmv-βNS-FC plasmid by PCR

then cutted by two endonucleases and directional cloned to the pAd track-cmv vector.Afterward

the correct recombinant was linearized by PmeI

following co-transformation with the backbone vector pAd Easy-1 in E.coli BJ5183.The homologous recombinant was linearized by PacI

and then transfected into HEK293 cell line to pack the adenovirus.Results:Through PCR

endonuclease cutting and gene sequencing

the target gene was verified to be correctly cloned in adenovirus vector.The high expression of green fluorescence protein in 293 cell line was found under fluorescent microscope.Conclusion:The recombinant adenovirus vector containing human β-catenin gene was successfully constructed and the adenovirus was packaged in HEK293 cell line.

关键词

β-catenin基因腺病毒载体Wnt/Wingless信号通路肿瘤

Keywords

β-cateninadenovirus vectorWnt/Wingless signal pathwaytumor

references

Views

166

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Advances in metabolic enzymes in the hexosamine biosynthetic pathway and OGT-mediated O-GlcNAc modification in tumors

Advances in and reflections on bacteria-mediated oncology therapy

Masses in maxillofacial region of 859 children:a retrospective analysis

Research progresses of the relationship between dioxygenases in kynurenine pathway and tumor

DETERMINATION OF ANTICANCER DRUG SENSITIVITY TEST BY THE MTT COLO RIMETRIC ASSAY FOR HUMAN SOLID TUMORS

Related Author

Yang Jiayao

Tang Ni

Wang Kai

Zining Wang

Ying Zhu

Yang Luo

Wei Gu

Li Liqi

Related Institution

Key Laboratory of Molecular Biology of Infectious Diseases，Ministry of Education，Chongqing Medical University

Center for Intelligent Testing and Molecular Medicine， School of Medicine， Chongqing University

Department of Stomatology,the Children’s Hospital,Chongqing Medical University

Department of Gastrointestinal Surgery,the Second Affiliated Hospital of Nanchang University

the Second Affiliated Medical College of Nanchang University

⁰