Yeramian P, Madaule P, Tavitian A
C R Acad Sci III. 1985;300(7):255-60.
We describe here an infectious eucaryotic expression vector derived from Moloney Murine Leukemia (Mo-MuLV) provirus and recombined in plasmid pBR 322, for the expression of eucaryotic genes. Upstream of the cloning sites lie the 5' LTR and 700 bp of the gag sequences containing the splicing and encapsidation signals. Downstream of the cloning sites are situated the env gene and the 3' LTR containing the polyadenylation signal. So as to test the potential use of this vector, Herpes Simplex TK gene and E. Coli NeoR genes were cloned in the same transcriptional polarity as the viral LTRs. When DNA from the recombinant plasmid was transfected into mouse, rat, or human cell cultures, high yields of TK+ or NeoR colonies were obtained. Recombinant plasmids constructed with TK or NeoR genes in the opposite polarity failed to produce drug resistant colonies. Cotransfection with DNA of the Mo-MuLV competent helper provirus led to the rescue of chimeric virus capable of transmitting drug resistance.
我们在此描述一种源自莫洛尼鼠白血病(Mo-MuLV)前病毒并重组于质粒pBR 322中的传染性真核表达载体,用于真核基因的表达。克隆位点上游是5'长末端重复序列(LTR)和包含剪接及包装信号的700 bp gag序列。克隆位点下游是env基因和包含聚腺苷酸化信号的3' LTR。为测试该载体的潜在用途,将单纯疱疹病毒TK基因和大肠杆菌NeoR基因以与病毒LTR相同的转录极性进行克隆。当将重组质粒的DNA转染到小鼠、大鼠或人类细胞培养物中时,获得了高产的TK+或NeoR菌落。用极性相反的TK或NeoR基因构建的重组质粒未能产生耐药菌落。与有活性的Mo-MuLV辅助前病毒DNA共转染导致能够传递耐药性的嵌合病毒的拯救。