Young J A, Bates P, Varmus H E
Department of Microbiology and Immunology, University of California School of Medicine, San Francisco 94143.
J Virol. 1993 Apr;67(4):1811-6. doi: 10.1128/JVI.67.4.1811-1816.1993.
We used a genetic strategy to isolate the chicken gene believed to encode the receptor for subgroup A avian leukosis and sarcoma viruses (ALSV-A). Chicken genomic DNA was transfected into monkey COS-7 cells, and two independent primary transfectants susceptible to ALSV-A infection were identified by using ALSV-A vectors containing a hygromycin B resistance gene. A second round of transfection and selection in mouse BALB/3T3 fibroblasts again led to isolation of a transfectant susceptible to infection by ALSV-A. Plasmid DNA sequences linked to chicken DNA during the primary transfection segregated with chicken DNA in the secondary transfectant and served as a molecular tag to clone the gene conferring susceptibility. Expression of the cloned gene in mouse BALB/3T3 cells conferred susceptibility to infection by ALSV-A but not by ALSV-B. Therefore the cloned gene most probably represents the tv-a locus, the genetically defined receptor gene for ALSV-A.
我们采用遗传策略来分离被认为编码A亚群禽白血病和肉瘤病毒(ALSV-A)受体的鸡基因。将鸡基因组DNA转染到猴COS-7细胞中,通过使用含有潮霉素B抗性基因的ALSV-A载体,鉴定出两个对ALSV-A感染敏感的独立原代转染子。在小鼠BALB/3T3成纤维细胞中进行第二轮转染和筛选,再次获得了对ALSV-A感染敏感的转染子。在原代转染过程中与鸡DNA相连的质粒DNA序列在二代转染子中与鸡DNA分离,并作为分子标签来克隆赋予易感性的基因。克隆基因在小鼠BALB/3T3细胞中的表达赋予了对ALSV-A而非ALSV-B感染的易感性。因此,克隆基因很可能代表tv-a基因座,即ALSV-A的遗传学定义的受体基因。