|本期目录/Table of Contents|

[1]杨文栋,苏 超,张春燕,等.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,2016,22(01):5-8.
 YANG Wen-dong,SU Chao,ZHANG Chun-yan,et al.Constrction and expression of recombinant pCMV-N-Tudor-SN.Mutants Flag plasmids[J].Journal of Tianjin Medical University,2016,22(01):5-8.
点击复制

重组pCMV-N-Tudor-SN点突变质粒的构建及表达(PDF)
分享到:

《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
22
期数:
2016年01
页码:
5-8
栏目:
基础医学
出版日期:
2016-01-20

文章信息/Info

Title:
Constrction and expression of recombinant pCMV-N-Tudor-SN.Mutants Flag plasmids
文章编号:
1006-8147(2016)01-0005-04
作者:
?杨文栋1苏 超2张春燕3赵亚丽1任媛媛3高星杰2杨 洁12何津岩1
(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)
Author(s):
YANG Wen-dong1 SU Chao2 ZHANG Chun-yan3 ZHAO Ya-li1 REN Yuan-yuan3 GAO Xing-jie2 YANG Jie12 HE Jin-yan1
(1. Department of Cell Biology;2. Research Center of Basic Medical Science;3. Department of Medical Biochemistry, Tianjin Medical University, Tianjin 300070, China )
关键词:
人类Tudor-SN蛋白pCMV-N-Flag重组质粒融合蛋白
Keywords:

line-height: 150%" align="left"> human Tudor-SNpCMV-N-Flagrecombinant plasmidfusion protein

分类号:
Q7
DOI:
-
文献标志码:
A
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。
Abstract:

Objective:To construct eukaryotic Flag expressing recombinant pCMV-N-Tudor-SN.Mutants-Flag. MethodsThe 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. ConclusionThe recombinant plasmids of pCMV-N-Tudor-SN.Mutants-Flag are constructed successfully, and may be useful for further study.

参考文献/References:

[1]Tong X, Drapkin R, Yalamanchili R, et al. The Epstein-Barr virus nuclear protein 2 acidic domain forms a complex with a novel cellular coactivator that can interact with TFIIE[J].Mol Cell Biol, 1995, 15(9):4735
[2]Callebaut I, Mornon J P. The human EBNA-2 coactivator p100: multidomain organization and relationship to the staphylococcal nuclease fold and to the Tudor protein involved in Drosophila melanogaster development[J].Biochem J,1997,321(Pt 1):125
[3]Su C, Zhang C, Tecle A, et al. Tudor staphylococcal nuclease (Tudor-SN), a novel regulator facilitating G1/S phase transition, acting as a co-activator of E2F-1 in cell cycle regulation[J].J Biol Chem,2015,290(11):7208
[4]Duan Z, Zhao X, Fu X, et al. Tudor-SN, a novel coactivator of peroxisome proliferator-activated receptor γ protein, is essential for adipogenesis[J].J Biol Chem,2014,289(12):8364
[5]Gao X, Ge L, Shao J, et al. Tudor-SN interacts with and co-localizes with G3BP in stress granules under stress conditions[J].FEBS Lett,2010,584(16):3525
[6]V?lineva T, Yang J, Palovuori R, et al. The transcriptional co-activator protein p100 recruits histone acetyltransferase activity to STAT6 and mediates interaction between the CREB-binding protein and STAT6[J].J Biol Chem,2005,280(15):14989
[7]Leverson J D, Koskinen P J, Orrico F C, et al. Pim-1 kinase and p100 cooperate to enhance c-Myb activity[J].Mol Cell,1998,2(4):417
[8]Gao X, Shi X, Fu X, et al. Human Tudor staphylococcal nuclease (Tudor-SN) protein modulates the kinetics of AGTR1-3’UTR granule formation[J].FEBS Lett,2014,588(13):2154
[9]Chiosea S, Jelezcova E, Chandran U, et al. Overexpression of dicer in precursor lesions of lung adenocarcinoma[J].Cancer Res, 2007, 67(5):2345
[10]Brito C C, Fachel A A, Vettore A L, et al. Identification of protein-coding and intronicnoncoding RNAs down-regulated in clear cell renal carcinoma[J].Mol Carcing,2008,47(10):757
[11]Tsuchiya N, Ochiai M, Nakashima K, et al. SND1, a component of RNA-induced silencing complex, is up-regulated in human colon cancers and implicated in early stage colon carcinogenesis[J].Cancer Res,2007,67(19):9568
[12]Kuruma H, Kamata Y, Takahashi H, et al. Staphylococcal nuclease domain-containing protein 1 as a potential tissue marker for prostate Cancer[J].Am J Pathol,2009,174(6):2044
[13]Low S H, Vasanth S, Larson C H, et al. Polycystin-1, STAT6, and P100 function in a pathway that transduces ciliary mechanosensation and is activated in polycystic kidney disease[J].Dev Cell,2006,10(1):57
[14] Sánchez-Martínez C, Gelbert LM, Lallena MJ, et al. Cyclin dependent kinase (CDK) inhibitors as anticancer drugs[J].Bioorg Med Chem Lett, 2015,25(17):3420
[15] Lam Y W, Trinkle-Mulcahy L. New insights into nucleolar structure and function[J].F1000Prime Rep,2015,7:48
[16] Tamura K. Development of cell-cycle checkpoint therapy for solid tumors[J].Jpn J Clin Oncol,2015,45(12):1097
[17] Wang X, Simpson E R, Brown K A. p53: Protection against tumor growth beyond effects on cell cycle and apoptosis[J].Cancer Res,2015,[Epub ahead of print]
[18]Jackson M A, Caputo N, Castle J R, et al. Stable liquid glucagon formulations for rescue treatment and bi-hormonal closed-loop pancreas[J].Curr Diab Rep,2012,12(6):705

相似文献/References:

备注/Memo

备注/Memo:

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



更新日期/Last Update: 2016-01-25