| 吴伟刚,刘妙娜,刘利平.CENPK基因过表达载体的构建及鉴定[J].中国肿瘤,2015,24(6):505-509. |
| CENPK基因过表达载体的构建及鉴定 |
| Construction and Identification of CENPK Gene Overexpressed Vector |
| 投稿时间：2014-09-10 |
| DOI：10.11735/j.issn.1004-0242.2015.06.A015 |
|
 |
| 中文关键词: CENPK基因 过表达 真核表达载体 肝癌FOCUS细胞 |
| 英文关键词:CENPK gene overexpression eukaryotic expression vector hepatoma FOCUS cells |
| 基金项目:国家自然科学基金（81372227）；深圳市科技计划项目（CXZZ20130515163643) |
|
| 摘要点击次数: 1608 |
| 全文下载次数: 1003 |
| 中文摘要: |
| 摘 要：［目的］ 构建着丝粒蛋白K (cetromere protein K，CENPK）基因过表达载体，并对其在肝癌FOCUS细胞中的表达进行鉴定。［方法］ ①从人肝癌组织中扩增出CENPK目的基因并将其克隆到pCMV-3Tag-3载体上，构建真核表达载体pCMV-CENPK；②利用脂质体法将构建好的pCMV-CENPK真核表达载体转染肝癌FOCUS细胞，并用G418筛选出CENPK基因稳定过表达的肝癌FOCUS细胞株；③通过Real-time PCR及Western Blot的方法对筛选出来的细胞进行鉴定。［结果］ ①经双酶切分析及测序验证，成功构建了真核表达载体pCMV-CENPK。②通过Real-time PCR及Western blot的方法鉴定，CENPK基因在筛选出来的肝癌FOCUS细胞株中成功过表达。［结论］ 成功构建真核表达载体pCMV-CENPK并使其在肝癌FOCUS细胞中稳定过表达，为进一步研究CENPK基因在肝癌发生发展中的作用奠定了基础。 |
| 英文摘要: |
| Abstract：［Purpose］ To construct CENPK gene overexpressed vector and to identify its expression in liver cancer FOCUS cells. ［Methods］ Amplify the CENPK gene from liver cancer tissues of human，and cloned it into pCMV-3Tag-3 vector，to construct eukaryotic expression vector pCMV-CENPK，then to transfect it into FOCUS cells by lipofection method，and to select it by G418. Cells selected by G418 were confirmed by Real-time PCR and Western Blot. ［Results］ Being verified with double endonuclease digestion and DNA sequencing，the eukaryotic expression vector pCMV-CENPK was successfully constructed. The FOCUS cells selected by G418 were successfully overexpressing CENPK gene，which was confirmed by Real-time PCR and Western blot. ［Conclusion］ The eukaryotic expression vector pCMV-CENPK is successfully constructed and was stably overexpressed on FOCUS cells，which lay the foundation for further study of CENPK gene on the role of carcinogenesis and progress of liver cancer. |
|
在线阅读
查看全文 查看/发表评论 下载PDF阅读器 |