摘要
在所有研究过的人体肿瘤组织或细胞中,p53似乎是突变频率最高的一个基因,研究和检测p53基因及其编码产物的变化将具有重要的意义。我们将野生型p53基因编码区3’端703bp的cD-NA片段插入到大肠杆菌表达载体pBV220中,得到了一个重组体表达质粒pRR33,经热诱导表达,用SDS-PAGE和Western印迹法证实p53蛋白多肽在大肠杆菌中得到了表达,它不仅可用于抗p53蛋白抗体的制备,而且也可用于对p53蛋白羧基端多肽功能的研究。
Mutations in the p53 gene are turning out to be the most common genetic alterations in human cancers. So it is very. important to study or detect the alterations of p53 gene in human cancers. We inserted a 703bp DNA fragment of wild-type p53 cDNA into a prokaryotic expression vector PBV220 and obtained a recombinant expression plasmid PRR33. By temperature indecent, SDS-PAGE and Western blotting, we demonstrated that a p53 peptide existed in E.coil containing the pRR33 plasmodia. This peptide can not only be used as an antigen to obtain antip53 antibody, but its function is also being studied.