人端粒酶逆转录酶启动子调控的携带胞嘧啶脱氨酶基因腺病毒载体的构建

doi:10.3877/cma.j.issn.1673-5250.2007.04.107

中华妇幼临床医学杂志(电子版) ›› 2007, Vol. 03 ›› Issue (04) : 202 -202. doi: 10.3877/cma.j.issn.1673-5250.2007.04.107

论著

人端粒酶逆转录酶启动子调控的携带胞嘧啶脱氨酶基因腺病毒载体的构建

黄淑华, 彭芝兰, 王和

广州市番禺区何贤纪念医院妇科(广州，511400)
四川大学华西第二医院妇产科

出版日期:2007-08-01

Construction of recombinant adenovirus containing cytosine deaminase gene driven by human telomerase reverse transcriptase promoter

Shu-hua HUANG, Zhi-lan PENG, He WANG

Hexian Memorial Hospital of Panyu, Panyu 511400, Guangzhou, China

Published:2007-08-01

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目的

构建由人端粒酶逆转录酶(human telomerase reverse transcriptase，hTERT)启动子驱动的含胞嘧啶脱氨酶自杀基因(cytosine deaminase gene，CD)的复制缺陷型腺病毒载体。

方法

首先构建带有hTERT启动子及CD基因的穿梭质粒pSU-TP-CD，然后与腺病毒骨架质粒pBHGE3共转染到293细胞，通过细胞内同源重组法获得复制缺陷性病毒。

结果

经PCR鉴定证明，成功构建了含有hTERT启动子及CD基因的重组腺病毒。

结论

成功构建了携带hTERT启动子和CD基因的复制缺陷型腺病毒Ad-hTERTp-CD，为下一步进行基因治疗研究奠定了基础。

关键词: 人端粒酶逆转录酶启动子, 胞嘧啶脱氨酶基因, 腺病毒载体

Objective

To construct recombinant adenovirus vectors in which the cytosine deaminase (CD) gene is under the control of the human telomerase reverse transcriptase (hTERT) promoter.

Methods

Recombined adenovirus of Ad-hTERTp-CD was constructed using homologous recombination in cells by transforming shuttle vector pSU-Tp-CD and adenovirus backbone vector pBHGE3 into 293 cells.

Results

Recombinant adenovirus vectors were confirmed by PCR analysis and DNA sequencing.

Conclusion

Replication-defective adenovirus vector in which the CD gene was driven by hTERT promoter was made successfully.

Key words: human telomerase reverse transcriptase (hTERT), cytosine deaminase gene (CD gene), adenovirus vector

图1　12号克隆酶切鉴定

图2　PCR扩增CD基因序列

图3　PCR扩增hTERT启动子序列

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[1]	黄淑华, 彭芝兰, 王和, 陈晓芸, 冯炜强. 人端粒酶逆转录酶启动子调控的胞嘧啶脱氨酶基因对宫颈癌细胞株的选择性杀伤作用研究[J/OL]. 中华妇幼临床医学杂志(电子版), 2010, 06(03): 172-174.
[2]	周华平, 彭和平, 刘启才. 腺病毒介导NK4基因和单核细胞趋化蛋白1基因共表达对胰腺癌细胞的影响[J/OL]. 中华普通外科学文献(电子版), 2013, 07(03): 165-169.

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Construction of recombinant adenovirus containing cytosine deaminase gene driven by human telomerase reverse transcriptase promoter