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抑制肿瘤细胞皮层肌动蛋白装配功能的重组质粒的构建和鉴定

Construction and identification of the recombinant plasmid of inhibition of tumor cell cortical actin assembly function
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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
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