Lipsick J S, Ibanez C E, Baluda M A
J Virol. 1986 Aug;59(2):267-75. doi: 10.1128/JVI.59.2.267-275.1986.
We demonstrated that molecular clones of the v-myb oncogene of avian myeloblastosis virus (AMV) can direct the synthesis of p48v-myb both in avian and mammalian cells which are not targets for transformation by AMV. To accomplish this, we constructed dominantly selectable avian leukosis virus derivatives which efficiently coexpress the protein products of the Tn5 neo gene and the v-myb oncogene. The use of chemically transformed QT6 quail cells for proviral DNA transfection or retroviral infection, followed by G418 selection, allowed the generation of cell lines which continuously produce both undeleted infectious neo-myb viral stocks and p48v-myb. The presence of a simian virus 40 origin of replication in the proviral plasmids also permitted high-level transient expression of p48v-myb in simian COS cells without intervening cycles of potentially mutagenic retroviral replication. These experiments establish that the previously reported DNA sequence of v-myb does in fact encode p48v-myb, the transforming protein of AMV.
我们证明,禽成髓细胞瘤病毒(AMV)的v-myb癌基因的分子克隆能够在禽类和哺乳动物细胞中指导p48v-myb的合成,而这些细胞并非AMV转化的靶细胞。为实现这一点,我们构建了具有显性选择功能的禽白血病病毒衍生物,其能高效共表达Tn5新霉素基因和v-myb癌基因的蛋白质产物。利用化学转化的QT6鹌鹑细胞进行原病毒DNA转染或逆转录病毒感染,随后进行G418筛选,从而产生能持续产生未缺失的感染性新霉素-myb病毒储备和p48v-myb的细胞系。原病毒质粒中猿猴病毒40复制起点的存在,也使得p48v-myb在猿猴COS细胞中能高水平瞬时表达,而无需经过可能具有诱变作用的逆转录病毒复制的中间循环。这些实验证实,先前报道的v-myb的DNA序列实际上编码了p48v-myb,即AMV的转化蛋白。