| 摘要: |
| 目的 构建癌基因p28GANK 缺失不同ankyrin序列的突变体,进一步研究p28GANK 的功能及在肝癌发生发展中的作用。方法 利用QuikChang Site-Directed Mutagenesis Kit 构建p28GANK 5个ankyrin 缺失突变体并检测各个突变体对p53的影响。 结果 构建出分别缺失第1个ankyrin(39~71aa)、第2个ankyrin (72~104aa)、第3个ankyrin(105~137aa)、第4个ankyrin(138~170aa)和第5个ankyrin(171~203aa)的突变体，琼脂糖凝胶电泳及蛋白免疫印迹法验证正确。野生型p28GANK 可促进p53降解,其他突变体对p53的表达无明显影响。 结论 成功构建了p28GANK 的5个突变体,证实p28GANK对p53的调控需要全部的ankyrin序列,为ankyrin的研究和p28GANK对肝癌的调控作用研究奠定了基础。 |
| 关键词: 癌基因蛋白质p28GANK 突变体 p53基因 |
| DOI：10.3724/SP.J.1008.2010.01049 |
| 投稿时间：2010-08-02修订日期：2010-09-21 |
| 基金项目:上海市自然科学基金(08ZR1405500). |
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| Construction of oncogene p28GANK mutants and their regulating effects on p53 |
| REN Yi-bin, FU Jing, CHEN Yao,,WANG Hong-yang* |
| (International Co-operation Laboratory on Signal Transduction, Eastern Hepatobiliary Surgery Institute, Second Military Medical University, Shanghai 200438, China) |
| Abstract: |
| Objective To construct the mutants of oncogene p28GANK with different ankyrin repeats deleted, so as to further study the potential role of p28GANK in hepatocellular carcinoma (HCC).Methods QuikChang Site-Directed Mutagenesis Kit was used to construct five p28GANK mutants with different ankyrin repeats deleted and their influences on p53 were also investigated. Results Five deletion mutants of p28GANK were as follows: 39-71aa, 72-104aa, 105-137aa, 138-170aa, and 171-203aa. Agarose gel electrophoresis and Western blotting analysis confirmed the correct construction of these mutants. Wild type p28GANK promoted degradation of p53, but the 5 mutants had no noticeable effects on expression of p53.Conclusion We have successfully constructed five p28GANK ankyrin repeat-deleted mutants. It is confirmed that the regulation of p53 needs the complete ankyrin repeats of p28GANK, which paves a way for further research on ankyrin and the role of p28GANK in HCC. |
| Key words: oncogene protein p28GANK mutants p53 gene |
