拟南芥耐盐相关基因AtSTK原核表达载体的构建及表达

doi:10.3969/j.issn.1000-7091.2013.02.007

华北农学报 ›› 2013, Vol. 28 ›› Issue (2): 38-41. doi: 10.3969/j.issn.1000-7091.2013.02.007

所属专题： 盐碱胁迫；生物技术；

拟南芥耐盐相关基因AtSTK原核表达载体的构建及表达

胡京蕊¹, 沈金宝², 李晶岚¹, 李晓旭¹, 赵宝存¹, 葛荣朝¹

1. 河北师范大学生命科学学院, 河北石家庄 050024;
2. 河北师范大学汇华学院, 河北石家庄 050024

收稿日期:2013-01-21 出版日期:2013-04-28
通讯作者: 葛荣朝(1974－)，男，河北故城人，教授，博士，主要从事植物抗逆基因的克隆与功能研究。
作者简介:胡京蕊(1983－)，女，河北隆尧人，助理农艺师，硕士，主要从事植物耐盐基因克隆和功能研究。
基金资助:
国家自然科学基金(30900104);河北省自然科学基金(C2009000271)

Prokaryotic Expression Vector Construction and Expression of Tolerance Relatedgene AtSTK in Arabidopsis

HU Jing-rui¹, SHEN Jin-bao², LI Jing-lan¹, LI Xiao-xu¹, ZHAO Bao-cun¹, gE Rong-chao¹

1. College of Life Science, Hebei Normal University, Shijiazhuang 050016, China;
2. Huihua College of Hebei Normal University, Shijiazhuang 050091, China

Received:2013-01-21 Published:2013-04-28

摘要/Abstract

摘要： 提取拟南芥总RNA,反转录获得cDNA后PCR扩增获得1 212 bp目的基因AtSTK。将该基因片段构建到PET-32a表达载体,转化大肠杆菌Rosetta菌,用IPTG进行诱导表达。该基因表达的最佳诱导条件为IPTG终浓度1mmol/L,诱导时间为4 h,此时融合蛋白表达量最高。该融合蛋白存在于包含体中,经包含体溶解、复性,最终纯化获得了AtSTK融合蛋白,为AtSTK蛋白的理化性质研究奠定了基础。

关键词: 拟南芥, 丝氨酸/苏氨酸蛋白激酶, 原核表达, 耐盐基因

Abstract: Total RNA was extracted from Arabidopsis leaves. 1212 bp AtSTKgene was amplified by RT-PCR and constructed into the expression vector PET-32a. Then the recombinant plasmid was transformed into E. coli Rosetta. The optimal induction conditions for AtSTKgene expression were 1 mmol/L IPTG and 4 hours induction. Under these conditions,the expression level of the fusion protein was highest. It was found that the form of the fusion protein was inclusion body. So the inclusion body was treated by dissolving and renaturation. Finally,the purified At-STK fusion protein was obtained. This work laid the foundation for the next study on the physical and chemical prop-erties of AtSTK protein.

Key words: Arabidopsis thaliana, Serine/threonine protein kinase, Prokaryotic expression, Salt tolerancegene

中图分类号:

胡京蕊, 沈金宝, 李晶岚, 李晓旭, 赵宝存, 葛荣朝. 拟南芥耐盐相关基因AtSTK原核表达载体的构建及表达[J]. 华北农学报, 2013, 28(2): 38-41. doi: 10.3969/j.issn.1000-7091.2013.02.007.

HU Jing-rui, SHEN Jin-bao, LI Jing-lan, LI Xiao-xu, ZHAO Bao-cun,gE Rong-chao. Prokaryotic Expression Vector Construction and Expression of Tolerance Relatedgene AtSTK in Arabidopsis[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(2): 38-41. doi: 10.3969/j.issn.1000-7091.2013.02.007.

http://www.hbnxb.net/CN/Y2013/V28/I2/38

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拟南芥耐盐相关基因AtSTK原核表达载体的构建及表达

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