| 重组pCMV-N-Tudor-SN点突变质粒的构建及表达 |
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| 引用本文： | ?杨文栋
?杨文栋,苏 超,张春燕,赵亚丽,任媛媛,高星杰,杨 洁,,何津岩.重组pCMV-N-Tudor-SN点突变质粒的构建及表达[J].天津医科大学学报,2016,0(1):5-8. |
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| 作者姓名： | ?杨文栋
?杨文栋 苏 超 张春燕 赵亚丽 任媛媛 高星杰 杨 洁 何津岩 |
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| 作者单位： | (天津医科大学1,.细胞生物学系;2.基础医学研究中心;3.医学生物化学与分子生物学系,天津 300070) |
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| 摘 要： | ?目的: 构建Tudor-SN蛋白的Serine426 (S426)、Serine781 (S781)、Threonine240 (T240) 和Threonine429 (T429)的点突变质粒，并使该重组质粒能够在HeLa细胞中融合表达。 方法:利用定点突变技术，对Tudor-SN蛋白进行S426A、S781A、T240A、T429A点突变，通过双酶切的方法获得Tudor- SN.Mutants 片段，最后将其连入到pCMV-N-Flag载体中。在HeLa细胞中转染该质粒，利用Western blot技术检测质粒表达效率。 结果:（1）重组质粒经双酶切鉴定，可以观察到载体与Tudor-SN.Mutants的条带。 （2）转染突变质粒后可看出HeLa细胞中有Flag蛋白表达。结论:质粒构建成功，可以用于下一步科学研究使用。
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| 关 键 词： | 人类Tudor-SN蛋白 pCMV-N-Flag 重组质粒 融合蛋白 |
Constrction and expression of recombinant pCMV-N-Tudor-SN.Mutants Flag plasmids |
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| YANG Wen-dong' target="_blank" rel="external">
YANG Wen-dong,SU Chao,ZHANG Chun-yan,ZHAO Ya-li,REN Yuan-yuan,GAO Xing-jie,YANG Jie,' target="_blank" rel="external">,HE Jin-yan.Constrction and expression of recombinant pCMV-N-Tudor-SN.Mutants Flag plasmids[J].Journal of Tianjin Medical University,2016,0(1):5-8. |
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| Authors: |
YANG Wen-dong" target="_blank">YANG Wen-dong' target="_blank" rel="external">
YANG Wen-dong SU Chao ZHANG Chun-yan ZHAO Ya-li REN Yuan-yuan GAO Xing-jie YANG Jie " target="_blank">' target="_blank" rel="external"> HE Jin-yan |
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| Institution: | （1. Department of Cell Biology；2. Research Center of Basic Medical Science；3. Department of Medical Biochemistry, Tianjin Medical University, Tianjin 300070, China ) |
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| Abstract: | Objective: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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| Keywords: | human Tudor-SN pCMV-N-Flag recombinant plasmid fusion protein |
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