酵母双杂交技术筛选NECL1胞内区的结合蛋白

基础医学与临床 ›› 2009, Vol. 29 ›› Issue (2): 119-123.

酵母双杂交技术筛选NECL1胞内区的结合蛋白

李晨陈涛阴彬龚燕华

中国医学科学院基础医学研究所北京协和医学院基础学院医学分子生物学国家重点实验室中国医学科学院基础医学研究所北京协和医学院基础学院医学分子生物学国家重点实验室中国医学科学院基础医学研究所北京协和医学院基础学院医学分子生物学国家重点实验室

收稿日期:2008-10-06 修回日期:2008-11-05 出版日期:2009-02-25 发布日期:2009-02-25
通讯作者: 李晨

Screening of proteins interacting with the cytoplasmic tail of NECL1 by yeast two-hybrid system

Chen LI, Tao CHEN, Bin YIN, Yan-hua GONG

National Laboratory of Medicine Molecular Biology, IBMS, CAMS，School of Basic Medicine PUMC National Laboratory of Medicine Molecular Biology, IBMS, CAMS，School of Basic Medicine PUMC National Laboratory of Medicine Molecular Biology, IBMS, CAMS，School of Basic Medicine PUMC

Received:2008-10-06 Revised:2008-11-05 Online:2009-02-25 Published:2009-02-25
Contact: Chen LI,

摘要/Abstract

摘要： 目的筛选人源膜表面黏附分子NECL1蛋白胞内区相互作用蛋白。方法构建含人NECL1蛋白胞内区氨基酸编码序列的诱饵质粒pGBKT7-NECL1C，对人胎脑cDNA文库进行酵母双杂交筛选。用GST pull down实验进行体外蛋白相互作用的验证。结果酵母双杂交阳性克隆测序后显示共存在9段不同序列（存在重复克隆）。比对氨基酸序列得到5个可能相互作用蛋白。通过GST pull down 实验验证了其中两个蛋白与NECL1胞内区的相互作用。结论应用酵母双杂交系统，获得了一些候选的NECL1胞内区相互作用蛋白。

关键词: 酵母双杂交, NECL1, 相互作用蛋白

Abstract: Objective To screen the proteins interact with the cytoplasmic tail of NECL1. Methods The cytoplasmic tail sequence of human NECL1 gene was inserted into the "bait" vector pGBKT7 in frame. The recombinant pGBKT7-NECL1C was then transformed in to yeast cells and followed by screening of human fetal brain cDNA library. Then the GST pull down assay was used to confirm the interactions between the proteins. Results Nine different sequences by sequencing the positive clones(There are repeated clones) and five alternative proteins were obtained by the amino acid sequence blast. Using the GST pull down analysis, we confirmed that two of them can interact with the cytoplasmic tail of NECL1 in vitro. Conclusion Using yeast two-hybrid system, we got some alternative proteins which may interact with the cytoplasmic tail of NECL1.

Key words: yeast two-hybrid, NECL1, interaction protein

中图分类号:

R739.41

李晨陈涛阴彬龚燕华. 酵母双杂交技术筛选NECL1胞内区的结合蛋白[J]. 基础医学与临床, 2009, 29(2): 119-123.

Chen LI; Tao CHEN; Bin YIN; Yan-hua GONG. Screening of proteins interacting with the cytoplasmic tail of NECL1 by yeast two-hybrid system[J]. Basic & Clinical Medicine, 2009, 29(2): 119-123.

[1]	景光旭, 王张鹏, 刘忠钰, 吕双, 孙红玉. 低氧预处理胎盘间充质干细胞减轻重症急性胰腺炎模型小鼠胰腺组织损伤[J]. 基础医学与临床, 2023, 43(9): 1346-1352.
[2]	高欣, 于永波, 王海存, 姜兴明, 王志东. 环状RNA核受体相互作用蛋白1对肿瘤发生发展的调控机制研究进展[J]. 基础医学与临床, 2023, 43(5): 827-832.
[3]	陈诗恺, 陈韭铭, 孙顺昌. C1orf194基因Ile122Asn突变致腓骨肌萎缩症的病理机制[J]. 基础医学与临床, 2021, 41(2): 159-164.
[4]	张文婷, 王瑾, 岳继萍, 靳文文, 张志楠, 焦向英. 硫氧还蛋白相互作用蛋白降低GLP-1促小鼠胰岛β细胞的增殖效应[J]. 基础医学与临床, 2020, 40(8): 1076-1082.
[5]	周鸿铭, 唐小牛, 郭伟, 吕业超, 姜玉新. Opa相互作用蛋白5在胰腺癌中的表达及其对PANC-1细胞增殖的影响[J]. 基础医学与临床, 2020, 40(6): 803-810.
[6]	范成艳郭正光郑直. 通过配体文库研究GIPC2 PDZ配体结合特点及其相互作用蛋白[J]. 基础医学与临床, 2016, 36(2): 186-190.
[7]	叶周恒刘文武孙学军. 程序性坏死机制与缺血再灌注损伤[J]. 基础医学与临床, 2014, 34(3): 414-417.
[8]	李嫄胡斯奇马素参邵晨牟一高友鹤. PDZK1和PDZK2结合特性的比较[J]. 基础医学与临床, 2010, 30(7): 703-707.
[9]	田瑞马素参杨涛高友鹤. 通过筛选随机多肽文库研究Veli3 PDZ结构域配体结合特性[J]. 基础医学与临床, 2009, 29(1): 14-18.