期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1983
卷号：80
期号：1
页码：56-59
DOI：10.1073/pnas.80.1.56
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：A cDNA library was constructed from sucrose-gradient-fractionated mRNA from SVT2, a mouse cell line transformed by simian virus 40. Polysomes containing the p53 messenger were specifically immunoprecipitated with monoclonal antibodies against the protein and used to prepare mRNA. This immunoselected mRNA, enriched 1,000-to 2,000-fold for p53-specific sequences, was used to make a cDNA probe and to screen the cDNA library. When approximately 10,000 colonies were screened by differential hybridization with probes made from immunoselected vs. nonenriched mRNA, a single clone was found that contained p53-specific sequences. The identity of this clone, termed pp53-208, was confirmed by the ability of its DNA to hybridize the mRNA coding for p53 (hybrid selection assay). When hybridized to a blot of EcoRI digested mouse DNA, the pp53-208 insert reacted with a single 3.3-kilobase band, suggesting that it is complementary to a single gene.
