[1]刘欣,张双喜,刘丹,等. 拟南芥蛋白激酶基因BIN2的原核表达纯化及其酶活性分析[J].西北林学院学报,2017,32(3):108-112.[doi:10.3969/j.issn.1001-7461.2017.03.20]
　LIU Xin,ZHANG Shuang-xi,LIU Dan,et al. Expression,Purification and Characterization of Arabidopsis BIN2[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2017,32(3):108-112.[doi:10.3969/j.issn.1001-7461.2017.03.20]
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 拟南芥蛋白激酶基因BIN2的原核表达纯化及其酶活性分析()
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《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]

卷:
第32卷
期数:
2017年第3期
页码:
108-112
栏目:
出版日期:
2017-05-31

文章信息/Info

Title:
 Expression,Purification and Characterization of Arabidopsis BIN2
文章编号:
1001-7461(2017)03-0108-05
作者:
 刘欣1; 2; 张双喜1; 2; 刘丹1; 2; 王晓峰1; 2; 陈少林1; 2*
 （1.西北农林科技大学 生命科学学院，陕西 杨陵 712100；2.西北农林科技大学 旱区生物质能研究中心，陕西 杨陵 712100）
Author(s):
 LIU Xin1; 2; ZHANG Shuang-xi1; 2; LIU Dan1; 2; WANG Xiao-feng1; 2; CHEN Shao-lin1; 2*
 （1.College of Life ScienceS,Northwest A&F University,Yangling,Shaanxi 712100,China; 2.Biomass Energy Center for Arid and Semi-arid Lands,Northwest A&F University,Yangling,Shaanxi 712100,China）
关键词:
 ; BIN2; SUMO; 原核表达; 激酶活性
Keywords:
 ; BIN2;  SUMO;  prokaryotic expression;  activity assay
分类号:
S718.46
DOI:
10.3969/j.issn.1001-7461.2017.03.20
文献标志码:
A
摘要:
 为了研究蛋白激酶BIN2（BR-insensitive-2）的底物及其在BR信号通路中的作用机制，通过One Step Cloning方法将BIN2基因克隆至SUMO表达载体，利用SUMO标签促进目的蛋白表达和折叠的特性，对BIN2蛋白进行原核表达和纯化，进而检测BIN2的激酶活性。结果表明，纯化得到的BIN2具有自身磷酸化活性，并测得其酶活大小和动力学曲线。为进一步研究BIN2蛋白的作用机制提供了基础材料。
Abstract:
 In order to study the target substrates of protein kinase BIN2 and its roles in the BR signaling pathway,we cloned BIN2 gene into the SUMO expression vector by one step cloning method.BIN2 was expressed and purified as a SUMO fusion protein,which facilitated folding and stabilization of heterologously expressed BIN2.Further activity analysis showed that purified BIN2 was functional and got autophosphorylated in vitro.In this study,the enzyme activity and kinetics curve of BIN2 were acquired as well.This laid a foundation for further analysis of the function and targets of BIN2.

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备注/Memo

备注/Memo:
 收稿日期：2017-02-05修回日期：2017-04-06
基金项目：西北农林科技大学引进人才启动项目（Z111021314）。
作者简介：刘欣，女，在读硕士，研究方向：植物细胞壁。E-mail：631433209@qq.com
*通信作者：陈少林，男，教授，研究方向：植物细胞壁。E-mail：slc1916@icloud.com
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