Harlow E, Williamson N M, Ralston R, Helfman D M, Adams T E
Mol Cell Biol. 1985 Jul;5(7):1601-10. doi: 10.1128/mcb.5.7.1601-1610.1985.
Three clones for the human tumor antigen p53 were isolated from a cDNA library prepared from A431 cells. One of these clones, pR4-2, contains the entire coding region for human p53. This clone directs the synthesis of a polypeptide with the correct molecular weight and immunological epitopes of an authentic p53 molecule in an in vitro transcription-translation reaction. Although the pR4-2 clone contains the coding region for p53, it is not a full-length copy of the human p53 mRNA. Northern analysis showed that the p53 mRNA is approximately 2,500 nucleotides long, whereas the pR4-2 insert is only 1,760 base pairs in length. Analysis of the DNA sequence of this clone suggests that the human p53 polypeptide has 393 amino acids. We compared the predicted amino acid sequence of the pR4-2 clone with similar clones for the mouse p53 and found long regions of amino acid homology between these two molecules.
从A431细胞制备的cDNA文库中分离出三个人类肿瘤抗原p53的克隆。其中一个克隆pR4 - 2包含人类p53的完整编码区。该克隆在体外转录 - 翻译反应中指导合成具有正确分子量和真实p53分子免疫表位的多肽。尽管pR4 - 2克隆包含p53的编码区，但它不是人类p53 mRNA的全长拷贝。Northern分析表明，p53 mRNA约2500个核苷酸长，而pR4 - 2插入片段仅1760个碱基对长。对该克隆的DNA序列分析表明，人类p53多肽有393个氨基酸。我们将pR4 - 2克隆预测的氨基酸序列与小鼠p53的类似克隆进行比较，发现这两个分子之间有长区域的氨基酸同源性。
