Cloning of human HA117 gene and preparation of itsrecombinant adenovirus vector

GUO Yu-xia; et al

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Cloning of human HA117 gene and preparation of itsrecombinant adenovirus vector

更新时间：2024-01-17

- Cloning of human HA117 gene and preparation of itsrecombinant adenovirus vector
- Journal of Chongqing Medical University Issue 6, Pages: 641-644(2008)
- 作者机构：
  
  1. 重庆医科大学附属儿童医院血液肿瘤科
  2. 重庆医科大学附属儿童医院血液肿瘤科,重庆,400014
- 作者简介：
- 基金信息：
- DOI：
  CLC： R73-3
- Published：2008，
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[1]郭玉霞,罗庆,徐酉华.耐药相关新基因HA117的克隆及重组腺病毒制备[J].重庆医科大学学报,2008(06):641-644.

GUO Yu-xia, et al. Cloning of human HA117 gene and preparation of itsrecombinant adenovirus vector. [J]. Journal of Chongqing Medical University (6):641-644(2008)
[1]郭玉霞,罗庆,徐酉华.耐药相关新基因HA117的克隆及重组腺病毒制备[J].重庆医科大学学报,2008(06):641-644. DOI：

GUO Yu-xia, et al. Cloning of human HA117 gene and preparation of itsrecombinant adenovirus vector. [J]. Journal of Chongqing Medical University (6):641-644(2008) DOI：

摘要

目的:构建表达耐药相关新基因HA117的重组腺病毒表达系统。方法:用基因工程技术将耐药相关新基因HA117的cDNA亚克隆至穿梭质粒pAdTrack-CMV上

然后在细菌内与pAdEasy同源重组

将重组腺病毒Ad5-HA117经脂质体转染293细胞包装、扩增腺病毒颗粒

通过荧光显微镜及流式细胞仪进行检测。结果:酶切鉴定及PCR结果证明耐药相关新基因HA117重组腺病毒载体构建成功

构建的耐药新基因HA117重组腺病毒载体的效价达到8.3×1011PFU/ml。结论:成功构建了表达ATRA相关耐药新基因HA117的重组腺病毒载体

为后续对HA117基因的相关研究奠定了基础。

Abstract

Objective:To construct the recombinant adenovirus expressing human HA117 gene.Methods:The eDNA of HA117 gene was subcloned into the shuttle plasmid pAdTrack-CMV and then the homologous recombinant adenovims genomie plasmid pAdEasy was cloned in bacteria.We tranfected the DNA of identified recombinant plasmid into 293 cells via liposome

made the package and amplification of adenovims

and determined them by fluorescence microscope and flow eytometry.Results:Restriction endonuelease and PCR analysis confirmed that the recombinant adenovims vector of HA117 gene was successfully constructed.The titer of the recombinant adenovims was 8.3×1011PFU/ml

and the expression of HA117 gene was observed.Conclusion:The recombinant adenovims expressing HA117 gene was successfully constructed

which paves the way for the following studies related to HA117.

关键词

基因HA117腺病毒转染

Keywords

Gene HA117adenovirustransfection

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

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