A binary adenoviral vector system for expressing high levels of the proapoptotic gene bax
A binary adenoviral vector system for expressing high levels of the proapoptotic gene bax
复制标题
用于表达高水平促凋亡基因bax的二元腺病毒载体系统
作者：
S. Kagawa;S. Pearson;L. Ji;Kai Xu;T. McDonnell;S. Swisher;J. Roth;B. Fang
bax基因在细胞凋亡的信号转导中起着关键作用，通过基因转移表达可能在治疗多种与细胞凋亡相关的疾病如癌症中有价值。然而，构建由组成型启动子驱动的表达bax基因的腺病毒载体一直是困难的，可能是因为该基因的高促凋亡活性。在这里，我们报告了一个系统，诱导的bax基因的表达安全的腺病毒介导的基因共转移。简而言之，该系统涉及含有人bax cDNA的腺病毒载体，该人bax cDNA由合成启动子驱动，该合成启动子由五个GAL 4结合位点和TATA盒（GT）组成。该载体在培养的哺乳动物细胞中表达最低背景水平的bax蛋白，从而防止包装细胞的凋亡，然而，通过将bax基因与表达反式激活因子融合蛋白GAL 4/VP 16的腺病毒载体一起沿着转移到靶细胞中，可以在体外和体内基本上诱导bax基因的表达。bax基因诱导人肺癌细胞和Balb/c小鼠肝细胞凋亡。我们的研究结果表明，这种GAL 4基因调控系统提供了一种替代方法来构建表达潜在毒性基因的病毒载体。
The bax gene plays a critical role in signaling apoptosis and expression through gene transfer may be valuable in the treatment of a variety of apoptosis-related diseases such as cancer. However, constructing an adenoviral vector expressing a bax gene driven by a constitutive promoter has been difficult, presumably because of the gene's high proapoptotic activity. Here we report a system that induces the expression of the bax gene safely by adenovirus-mediated gene cotransfer. Briefly, the system involves an adenoviral vector containing a human bax cDNA driven by a synthetic promoter consisting of five GAL4-binding sites and a TATA box (GT). This vector expresses a minimal background level of bax protein in cultured mammalian cells thus preventing apoptosis of packaging cells, however, expression of the bax gene can be induced substantially in vitro and in vivo by transferring it into target cells along with an adenoviral vector expressing the transactivator, fusion protein GAL4/VP16. Extensive apoptosis was observed after induction of the bax gene both in cultured human lung carcinoma cells and in the livers of Balb/c mice. Our results suggest that this GAL4 gene regulatory system provides an alternative approach to constructing viral vectors that express potentially toxic genes.
DOI：
10.1073/pnas.91.10.4407
发表时间：
1994-05-10
影响因子：
11.1
作者：
YANG, YP;NUNES, FA;WILSON, JM
通讯作者：
WILSON, JM
通过脂质体介导的转染和 PCR 分析产生和鉴定重组腺病毒。
影响因子：
2.7
作者：
Zhang,WW;Fang,X;Branch,CD;Mazur,W;French,BA;Roth,JA
通讯作者：
Roth,JA
DOI：
10.1073/pnas.94.25.13862
发表时间：
1997-12-09
影响因子：
11.1
作者：
Arai, H;Gordon, D;Nabel, GJ
通讯作者：
Nabel, GJ