Hauer C A, Getty R R, Tykocinski M L
Institute of Pathology, Case Western Reserve University, Cleveland, OH 44106.
Nucleic Acids Res. 1989 Mar 11;17(5):1989-2003. doi: 10.1093/nar/17.5.1989.
Epstein-Barr virus (EBV) episomal replicons offer an expeditious means for amplifying transfected genes in human cells. A panel of EBV episomes was constructed to assess the relative utility of five distinct eukaryotic promoter elements for high level and inducible gene expression in stably transfected human myeloid leukemia cells. The Rous sarcoma virus 3' long terminal repeat (LTR) was most highly suited for EBV episome-based gene expression, whereas the lymphopapilloma virus and the SV40 early regulatory elements exhibited substantially lower activities. Chemically responsive promoter elements, such as the SV40 early, human metallothionein IIA and rat GRP78 gene promoters, retained their inducibility when EBV episome-based. Differences in gene expression obtained with the episomes reflected differential promoter activity rather than significant variations in episome copy numbers per cell. These observations provide guidelines for the optimal design of EBV episomal expression vectors for human expression work.
爱泼斯坦-巴尔病毒(EBV)附加型复制子为在人类细胞中扩增转染基因提供了一种快速方法。构建了一组EBV附加体,以评估五种不同的真核启动子元件在稳定转染的人髓系白血病细胞中进行高水平和诱导型基因表达的相对效用。劳氏肉瘤病毒3'长末端重复序列(LTR)最适合基于EBV附加体的基因表达,而淋巴乳头瘤病毒和SV40早期调控元件的活性则显著较低。化学应答启动子元件,如SV40早期、人金属硫蛋白IIA和大鼠GRP78基因启动子,在基于EBV附加体时仍保持其诱导性。附加体获得的基因表达差异反映了启动子活性的差异,而非每个细胞附加体拷贝数的显著变化。这些观察结果为用于人类表达工作的EBV附加型表达载体的优化设计提供了指导。