Iguchi-Ariga S M, Itani T, Kiji Y, Ariga H
Institute of Medical Science, University of Tokyo, Japan.
EMBO J. 1987 Aug;6(8):2365-71. doi: 10.1002/j.1460-2075.1987.tb02513.x.
We have recently cloned a plasmid, pARS65, containing the sequences derived from mouse liver DNA which can autonomously replicate in mouse and human cells (Ariga et al., 1987). In this report, we show that replication of pARS65 in HL-60 cells can be inhibited by co-transfection with anti-c-myc antibody. In an in-vitro replication system using HL-60 nuclear extract, pARS65 functioned as a template. This in-vitro replication was also blocked by addition of anti-c-myc antibody. Specific binding activity of the c-myc product to pARS65 was detected by an immunobinding assay, suggesting that the c-myc protein promotes DNA replication through binding to the initiation site of replication. This has been substantiated using the antibody to help isolate a human DNA segment that can autonomously replicate in the cells.
我们最近克隆了一种质粒pARS65,它含有源自小鼠肝脏DNA的序列,能够在小鼠和人类细胞中自主复制(有贺等人,1987年)。在本报告中,我们表明,用抗c-myc抗体共转染可抑制pARS65在HL-60细胞中的复制。在使用HL-60核提取物的体外复制系统中,pARS65充当模板。添加抗c-myc抗体也会阻断这种体外复制。通过免疫结合试验检测到c-myc产物与pARS65的特异性结合活性,这表明c-myc蛋白通过与复制起始位点结合来促进DNA复制。使用该抗体帮助分离出一个能够在细胞中自主复制的人类DNA片段,这一观点得到了证实。