Identification of interacting protein of FOXP3 by yeast two-hybrid system

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

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Identification of interacting protein of FOXP3 by yeast two-hybrid system

更新时间：2025-01-15

- Identification of interacting protein of FOXP3 by yeast two-hybrid system
- Journal of Army Medical University Vol. 30, Issue 24, Pages: 2258-2261(2008)
- 作者机构：
  
  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
- 作者简介：
- 基金信息：
- DOI：
  CLC： R341;R392.2;R392-33
- Published：2008，
  
  Published Online：30 December 2008，
- 稿件说明：
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ZHOU LI-NA, WU JUN, HE WEI-FENG, et al. Identification of interacting protein of FOXP3 by yeast two-hybrid system. [J]. Journal of army medical university, 2008, 30(24): 2258-2261.
DOI：

ZHOU LI-NA, WU JUN, HE WEI-FENG, et al. Identification of interacting protein of FOXP3 by yeast two-hybrid system. [J]. Journal of army medical university, 2008, 30(24): 2258-2261. DOI：

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.

关键词

基因克隆Foxp3酵母双杂交

Keywords

genecloneFoxp3yeast two hybrid

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