Molecular cloning of a cDNA specific for the murine p53 cellular tumor antigen.
Molecular cloning of a cDNA specific for the murine p53 cellular tumor antigen.
复制标题
鼠 p53 细胞肿瘤抗原特异性 cDNA 的分子克隆。
DOI：
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发表时间：
1983
影响因子：
11.1
通讯作者：
A. Levine
中科院分区：
文献类型：
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作者：
M. Oren;A. Levine
以猴病毒40转化的小鼠细胞系SVT2的蔗糖梯度分离的mRNA为模板，构建了一个cDNA文库。含有P53信使的多聚体与抗P53蛋白的单抗进行特异性免疫沉淀，并用于制备mRNA。这种免疫选择的mRNA1,000-2,000倍的P53特异性序列，被用来制作cDNAs探针和筛选cDNA库。当用免疫选择的和非浓缩的mRNA制成的探针通过差异杂交筛选大约10,000个克隆时，发现了一个包含P53特异性序列的克隆。该克隆被命名为pp53-208，其DNA与编码p53的mRNA杂交(杂交选择试验)的能力证实了该克隆的身份。当与EcoRI消化的小鼠DNA的印迹杂交时，pp53-208插入片段与单个3.3kb的条带反应，表明它对单个基因是互补的。
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.
DOI：
10.1073/pnas.79.2.436
发表时间：
1982-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
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作者：
CAMPISI, J;MEDRANO, EE;PARDEE, AB
通讯作者：
PARDEE, AB