Spandidos D A, Wilkie N M
EMBO J. 1983;2(7):1193-9. doi: 10.1002/j.1460-2075.1983.tb01566.x.
The bovine papillomavirus (BPV-1), Moloney murine sarcoma virus (MoMuSV) and simian virus 40(SV40) genomes have been shown to contain sequences termed 'enhancers' which activate the expression of linked genes. DNA fragments containing these three enhancers have been inserted into recombinant plasmids upstream from the herpes simplex virus thymidine kinase (tk) gene, and their effect on tk expression monitored. Two types of assay have been used. Firstly, the ability of recombinant plasmids to transform TK- recipient cells to a TK+ phenotype was measured. Secondly, the amount of tk-specific RNA and TK enzyme activity transiently expressed after DNA transfection was determined. Both types of assay gave similar results. The enhancers increased tk gene expression by regulating the amount of full length tk mRNA present shortly after transfection independent of gene copy number. Furthermore, marked species specificity in the relative efficiencies of different enhancers was observed, including that of the BPV-1 enhancer for the first time. The MoMuSV enhancer showed preference for murine fibroblasts, while the papillomavirus enhancer showed a marked preference for bovine cells. In contrast, the SV40 enhancer gave the same relative increase in tk gene expression in the murine, rat, bovine and human cells tested.
牛乳头瘤病毒(BPV-1)、莫洛尼鼠肉瘤病毒(MoMuSV)和猴病毒40(SV40)的基因组已被证明含有被称为“增强子”的序列,这些序列可激活相连基因的表达。含有这三种增强子的DNA片段已被插入到单纯疱疹病毒胸苷激酶(tk)基因上游的重组质粒中,并监测它们对tk表达的影响。已使用了两种类型的检测方法。首先,测定重组质粒将TK-受体细胞转化为TK+表型的能力。其次,测定DNA转染后瞬时表达的tk特异性RNA的量和TK酶活性。两种检测方法都得到了相似的结果。增强子通过调节转染后不久存在的全长tk mRNA的量来增加tk基因的表达,而与基因拷贝数无关。此外,观察到不同增强子的相对效率存在明显的物种特异性,首次包括BPV-1增强子的物种特异性。MoMuSV增强子对鼠成纤维细胞表现出偏好,而乳头瘤病毒增强子对牛细胞表现出明显偏好。相比之下,SV40增强子在测试的鼠、大鼠、牛和人细胞中使tk基因表达产生相同的相对增加。