[1]周丽娜,吴军,贺伟峰,等.酵母双杂交系统筛选FOXP3的相互作用蛋白[J].陆军军医大学学报(原第三军医大学学报),2008,30(24):2258-2261.
 ZHOU Li-na,WU Jun,HE Wei-feng,et al.Identification of interacting protein of FOXP3 by yeast two-hybrid system[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(24):2258-2261.
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第24期
页码:
2258-2261
栏目:
论著
出版日期:
2008-12-30

文章信息/Info

Title:
Identification of interacting protein of FOXP3 by yeast two-hybrid system
作者:
周丽娜吴军贺伟峰罗高兴陈希炜柏干苹袁顺宗张小容胡晓红
第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室,疾病蛋白质组重庆市市级重点实验室
Author(s):
ZHOU Li-na WU Jun HE Wei-feng LUO Gao-xing CHEN Xi-wei BAI Gan-ping YUAN Shun-zong ZHANG Xiao-rong HU Xiao-hong
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burns, Southwest Hospital, Third Military Medical University, Chongqing Key Laboratory for Proteomics of Diseases, Chongqing 400038, China
关键词:
基因克隆FOXP3酵母双杂交
Keywords:
gene clone FOXP3 yeast two hybrid
分类号:
R341;R392-33;R392.2
文献标志码:
A
摘要:
目的    利用酵母双杂交系统筛选与人FOXP3相互作用的蛋白。    方法    应用酵母双杂交系统,将成功构建的酵母诱饵表达载体pGBKT7-FOXP3,与转化了成人肝cDNA文库的酵母菌Y187接合,筛选与FOXP3相互作用的蛋白,通过一对一的回复性杂交实验排除假阳性,并对阳性克隆进行序列测定和生物信息学分析。    结果    经过酵母双杂交共获得最终确认3个阳性克隆,测序后经生物信息分析为肿瘤蛋白D52、分裂因子3b及未知蛋白。    结论    成功应用酵母双杂交系统筛选出与FOXP3相互作用的3个蛋白,它们可能与Treg细胞的发育分化及其功能有关。
Abstract:
Objective    To screen the proteins interacting with FOXP3 in yeast two-hybrid system.     Methods    The “bait plasmid” pGBKT7 (named as pGBKT7-FOXP3) was constructed successfully. Using FOXP3 as bait, a human liver cDNA library was screened and the proteins interacting with FOXP3 were searched. The false positive clones were discarded by one to one yeast two-hybrid system, and the positive clones were sequenced and analyzed by bioinformatic methods.     Results    The bait plasmid pGBKT7-FOXP3 was constructed successfully and there was no self-activation or toxicity in AH109. Three proteins had been found in our system to be able to interact with FOXP3. They were tumor protein D52, splicing factor 3b subunit 1 and one hypothetical protein.     Conclusion    FOXP3 interacts with tumor protein D52, splicing factor 3b subunit 1 and one hypothetical protein, all of which may interfere in cell metabolism and function of T cell.

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更新日期/Last Update: 2008-11-12