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作者以Hela细胞液体系,研究大鼠细胞色素P450IA-1(P 450 C)基因在4S多环芳香烃(PAH)结合蛋白存在下的体外转录作用。用含大鼠细胞色素P 450 IA1的-880至外显子的第1000碱基对((bp)的质粒pA9[长约1.94千碱基对(kb)],经内切酶切割成不同大小的片段为模板。以插有腺病毒-2大部晚期启动子(Ad-2)的质粒PHML-1作为阳性对照。所得结果显示Hela细胞体系能按模板准确地转录出预期的产物。加入4S PAH-结合蛋白使pA9模板的转录增加2～3倍,而对PHML-1的转录无增强作用,说明4S PAH-结合蛋白是专一加强细胞色素P450IA1基因转录的正调节因素。推测4S PAH-结合蛋白也可能诱导大鼠体内细胞色素P 450 IA1基因的表达。
In order to study the in vitro transcription of rat cytochrome P450IA-1 (P 450 C) gene in the presence of 4S polycyclic aromatic hydrocarbons (PAH) -binding proteins, Hela cell line was used to investigate the in vitro transcription of rat cytochrome P450IA-1 (P 450 C) Plasmid pA9 [about 1.94 kilobases (kb)] containing the base-1000 bp (-880 bp) of exon of rat cytochrome P450 IA1 was digested with endonucleases to different sizes As the template.The plasmid PHML-1 with adenovirus-2 large part of late promoter (Ad-2) was used as a positive control.The results showed that Hela cell system can accurately transcribe the expected product according to the template.4S PAH-binding protein increased the transcription of pA9 template 2 to 3 times, but did not enhance the transcription of PHML-1, indicating that 4S PAH-binding protein is a positive regulator to specifically enhance the transcription of cytochrome P450IA1. Binding proteins may also induce the expression of the cytochrome P 450 IA1 gene in rats.