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; " align="left"> human Tudor-SN; pCMV-N-Flag; recombinant plasmid; fusion proteinObjective:To construct eukaryotic Flag expressing recombinant pCMV-N-Tudor-SN.Mutants-Flag. Methods:The Serine426(S426)、Serine781(S781)、Threonine240(T240)and Threonine429(T429) of Tudor-SN were transformed into Alanine by site- directed mutagenesis technique. Then the Tudor-SN.Mutants were obtained by restricting double enzyme digestion, and then inserted into pCMV-N-Flag vector. The recombinant plasmids were transfected into HeLa and observed by Western Blot. Results:(1) The vector and Tudor-SN.Mutants could be observed by restricting double enzyme digestion. (2) Flag was expressed by HeLa which was transfected with recombinant plasmid. Conclusion:The recombinant plasmids of pCMV-N-Tudor-SN.Mutants-Flag are constructed successfully, and may be useful for further study.
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基金项目 国家杰出青年基金资助项目(No.31125012);教育部“创新团队发展计划”( No.IRT13085);国家自然科学基金资助项目(No.31170830、31370749、31571380);天津市应用基础与前沿技术研究计划(青年基金项目)(No.15JCQNJC09900)
作者简介 杨文栋(1989-),男,硕士在读,研究方向:肿瘤免疫;通信作者:何津岩,Email:hejinyan@tijmu.edu.cn。