Oren M, Bienz B, Givol D, Rechavi G, Zakut R
EMBO J. 1983;2(10):1633-9. doi: 10.1002/j.1460-2075.1983.tb01637.x.
Three cDNA clones, corresponding to two non-overlapping regions of the mRNA coding for the mouse p53 cellular tumor antigen, were isolated and characterized. In hybridization-selection assays, these clones were capable of selectively binding p53 mRNA, as demonstrated by in vitro translation and immunoprecipitation with anti-p53 monoclonal antibodies. The p53 mRNA appeared to be the only messenger species specifically selected by these clones. The size of the p53 mRNA was found to be approximately 2 kb, and its levels to vary substantially among different types of transformed cells. Evidence was found for the existence of two distinct p53-specific genes in mouse genomic DNA. Two partially overlapping recombinant phage clones were obtained, both derived from the same p53-specific genomic DNA region. The orientation of the various cDNA clones relative to that of the p53 mRNA was established by S1 analysis and the relationship between the cDNA clones and the genomic ones was determined by comparative restriction enzyme mapping and nucleic acid hybridization.
分离并鉴定了三个与编码小鼠p53细胞肿瘤抗原的mRNA的两个非重叠区域相对应的cDNA克隆。在杂交选择试验中，如体外翻译和用抗p53单克隆抗体进行免疫沉淀所证明的，这些克隆能够选择性地结合p53 mRNA。p53 mRNA似乎是被这些克隆特异性选择的唯一信使种类。发现p53 mRNA的大小约为2 kb，其水平在不同类型的转化细胞中差异很大。在小鼠基因组DNA中发现了两个不同的p53特异性基因存在的证据。获得了两个部分重叠的重组噬菌体克隆，二者均来自同一p53特异性基因组DNA区域。通过S1分析确定了各种cDNA克隆相对于p53 mRNA的方向，并通过比较限制性酶切图谱和核酸杂交确定了cDNA克隆与基因组克隆之间的关系。
