Berland R, Chasin L A
Department of Biological Sciences, Columbia University, New York 10027.
Nucleic Acids Res. 1988 Dec 23;16(24):11573-90. doi: 10.1093/nar/16.24.11573.
The tissue-specific expression of the liver-specific rat albumin gene promoter was examined after transfer to various hepatic and non-hepatic cell lines. A 402 base pair sequence from the albumin gene 5' flank enabled a fused reporter chloramphenicol acetyltransferase gene to be expressed in rat hepatoma cell lines but not in fibroblast lines or dedifferentiated hepatoma cells. However, when this same construct was analyzed in permanently transfected cell populations, it was expressed equally well in differentiated and dedifferentiated hepatoma cells and in two of three fibroblast lines tested. The inappropriate expression of the albumin promoter was also seen using the HSV tk gene and the E. coli gpt gene as reporters, and when assayed by colony formation in HAT medium (tk gene) or by S1 protection of transcripts in cotransfected populations (tk and gpt genes). These results show that gene regulatory elements can behave differently in transient vs. stable transfections, and suggest that chromosomal integration can provide long range positive influences on gene expression.
将肝脏特异性大鼠白蛋白基因启动子转入各种肝细胞系和非肝细胞系后,检测了其组织特异性表达情况。白蛋白基因5'侧翼的一段402个碱基对的序列,能使融合的报告氯霉素乙酰转移酶基因在大鼠肝癌细胞系中表达,但在成纤维细胞系或去分化肝癌细胞中不表达。然而,当在永久转染的细胞群体中分析相同构建体时,它在分化和去分化的肝癌细胞以及所测试的三个成纤维细胞系中的两个中表达效果相同。使用单纯疱疹病毒胸苷激酶(HSV tk)基因和大肠杆菌黄嘌呤 - 鸟嘌呤磷酸核糖转移酶(gpt)基因作为报告基因时,也观察到白蛋白启动子的异常表达,并且通过在HAT培养基中进行集落形成分析(tk基因)或通过对共转染群体中的转录本进行S1核酸酶保护分析(tk和gpt基因)时也观察到。这些结果表明,基因调控元件在瞬时转染和稳定转染中的行为可能不同,并表明染色体整合可以对基因表达产生远距离的正向影响。