摘要
目的 构建乌拉尔甘草基本螺旋-环-螺旋(GubHLH)转录因子的原核表达载体并进行原核表达及鉴定。方法 从乌拉尔甘草根中提取总mRNA,反转录得到cDNA后PCR扩增获得GubHLH基因全长,构建原核表达载体pET28a-GubHLH,分析GubHLH表达的最佳IPTG浓度、温度和时间条件。结果 获得的重组质粒pET28a-GubHLH在大肠杆菌E.coli BL21(DE3)中的最佳诱导条件为37℃下用0.2 mmol/L的IPTG诱导目的蛋白3 h。结论 成功构建GubHLH蛋白原核表达体系,并获得重组质粒pET28a-GubHLH在大肠杆菌E.coli BL21(DE3)中的最佳诱导条件。
Objective To construct a prokaryotic expression vector of the basic helix loop helix(GubHLH)transcription factor in licorice(Glycyrrhiza uralensis Fisch.)and prokaryotic expression was performed and identified.Methods The total mRNA was extracted from licorice root,and reverse transcription polymerase chain reaction was performed to amplify the full length of GubHLH gene.The prokaryotic expression vector pET28a-GubHLH was constructed and the optimal IPTG concentration,temperature,and time conditions for GubHLH expression were analyzed.Results The optimal induction condition combination of obtained recombinant plasmid pET28a-GubHLH in E.coli BL21(DE3)is to induce the target protein with 0.2 mmol/L IPTG at 37 ℃ for 3 hours.Conclusion The prokaryotic expression system of GubHLH protein is constructed successfully.The optimal induction condition combination of obtained recombinant plasmid pET28a-GubHLH in E.coli BL21(DE3)is confirmed.
作者
张艺朦
付业繁
赫苯玎
陈晓芳
孙欣
李欣
方圆
ZHANG Yimeng;FU Yefan;HE Bending;CHEN Xiaofang;SUN Xin;LI Xin;FANG Yuan(Jilin Medical University,Jilin City,Jilin Province 132013,China)
出处
《吉林医药学院学报》
2025年第1期15-18,共4页
Journal of Jilin Medical University
基金
吉林医药学院大学生创新创业训练计划项目(2023CXXL028)。
关键词
甘草
原核表达
bHLH基因
licorice(Glycyrrhiza uralensis Fisch.)
prokaryotic expression
bHLH gene
