Dennis E, Berg P
Nucleic Acids Res. 1985 Nov 25;13(22):7945-57. doi: 10.1093/nar/13.22.7945.
The promoter segment of a plant gene (maize alcohol dehydrogenase 1 (Adh 1)) has been fused to two bacterial reporter genes, Ecogpt (1) and neo (2), in pSV2-derived vectors and introduced into cultured mammalian cells by DNA transfection. The pAdh1-gpt plasmids transformed the recipient cells for resistance to mycophenolic acid plus xanthine (3) and the analogous pAdh1-neo plasmid transformed cells to G418 resistance (2). S1 analysis of transient transfections of CV1 cells with various derivatives of pAdh1-gpt confirmed that production of gpt mRNA is initiated at the Adh1 promoter at and near the same site used in transcription of the intact Adh1 gene in maize. Moreover, expression of the Adh1 promoter was increased 10-20 fold if the SV40 early region enhancer sequence was included in the same molecule.
植物基因(玉米乙醇脱氢酶1(Adh 1))的启动子片段已与两个细菌报告基因Ecogpt(1)和neo(2)在源自pSV2的载体中融合,并通过DNA转染导入培养的哺乳动物细胞。pAdh1 - gpt质粒使受体细胞转化为对霉酚酸加黄嘌呤具有抗性(3),类似的pAdh1 - neo质粒使细胞转化为对G418具有抗性(2)。用pAdh1 - gpt的各种衍生物对CV1细胞进行瞬时转染的S1分析证实,gpt mRNA的产生在玉米完整Adh1基因转录所用的相同位点及附近的Adh1启动子处起始。此外,如果同一分子中包含SV40早期区域增强子序列,Adh1启动子的表达会增加10 - 20倍。