摘要
目的:构建一个具有拮抗肌动蛋白装配功能的重组质粒EGFP-N2/CA,为观察该基因片段对于肿瘤细胞的侵袭转移能力的影响。方法:通过PCR扩增获得目的片段N-WASP-CA,通过双酶切和连接将其插入荧光表达载体EGFP-N2获得重组质粒。结果:PCR扩增后获得和预期一致的特异性DNA片段,通过PCR、双酶切和测序证实该片段已正确插入表达载体。结论:成功获得重组质粒EGFP-N2/CA。
Objective: To construct a recombinant plasmid EGFP-N2/CA with the function of inhibiting cortical aetin assembly,so as to make a preparation for the further study on the invasion and metastasis of tumor cells. Methods: Gene(N-WASP-CA) was amplified through polymerase chain reaction (PCR), and then was inserted into the florescence expression vector-EGFP-N2, after double digestion and ligation with restriction endoneleases and ligase. Results: The correct CA clone was obtained and it was confirmed that CA was inserted into the expression vector correctly by PCR, double digestion identification and seqeneing. Conclusion: The recombinant plasmid EGFP-N2/CA is successflly constructed.
出处
《交通医学》
2009年第5期456-458,共3页
Medical Journal of Communications
基金
国家自然科学基金(基金编号30771126)
江苏省自然科学基金(基金编号BK2006058)资助
关键词
皮层肌动蛋白装配
重组质粒
N-WASP
cortical actin assembly
recombinant plasmid
N-Wiskott-Aldrich synclorme protein