拟南芥耐盐相关基因AtSTK原核表达载体的构建及表达  

Prokaryotic Expression Vector Construction and Expression of Tolerance Related Gene AtSTKin Arabidopsis

作　　者：胡京蕊[1] 沈金宝[2] 李晶岚[1] 李晓旭[1] 赵宝存[1] 葛荣朝[1] 

机构地区：[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

出　　处：《华北农学报》2013年第2期38-41,共4页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金(30900104);河北省自然科学基金(C2009000271)

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。Total RNA was extracted from Arabidopsis leaves.1 212 bp AtSTK gene 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 AtSTK gene 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 AtSTK fusion protein was obtained.This work laid the foundation for the next study on the physical and chemical properties of AtSTK protein.

关 键 词：拟南芥 丝氨酸 苏氨酸蛋白激酶 原核表达 耐盐基因 

分 类 号：Q78[生物学—分子生物学]