苹果响应钙信号转录因子MdbHLH2原核表达分析  

Prokaryotic expression analysis of calcium signal transcription factor MdbHLH2 in apple

作　　者：苟莎莎 安欣 张玮玮 胡康棣 张华 姚改芳 GOU Shasha;AN Xin;ZHANG Weiwei;HU Kangdi;ZHANG Hua;YAO Gaifang(School of Food and Biological Engineering,Hefei University of Technology,Hefei 230601,China;College of Horticulture Science and Engineering,Shandong Agricultural University,Tai’an 271001,China)

机构地区：[1]合肥工业大学食品与生物工程学院,安徽合肥230601 [2]山东农业大学园艺科学与工程学院,山东泰安271001

出　　处：《合肥工业大学学报(自然科学版)》2025年第2期232-235,共4页Journal of Hefei University of Technology:Natural Science

基　　金：国家重点研发计划资助项目(2019YFD1000103);国家自然科学基金资助项目(31901993);安徽省自然科学基金资助项目(1908085MC72)。

摘　　要：转录因子MdbHLH2能响应钙信号且影响苹果采后贮藏的性能。为进一步探究其结构和功能,文章利用聚合酶链式反应(polymerase chain reaction,PCR)扩增MdbHLH 2基因的编码序列(coding sequence,CDS)全长,连接至pMAL-c2X原核表达载体,并将重组质粒转入表达菌株Rosetta(DE3);利用异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达融合MBP标签的MdbHLH2重组蛋白,并用聚丙烯酰胺凝胶电泳(PAGE)技术检测出MdbHLH2-MBP的重组表达蛋白。该结果为进一步研究MdbHLH2的结构和功能提供了重要依据。Transcription factor MdbHLH2 can respond to calcium signal and affect the postharvest storage performance of apple.In order to further explore its structure and function,the full length of MdbHLH 2 gene coding sequence(CDS)was amplified by polymerase chain reaction(PCR)and cloned into prokaryotic expression vector pMAL-c2X,and the recombinant plasmid was transformed into expression strain Rosetta(DE3).The recombinant MdbHLH2 protein fused with MBP tag was induced by isopropyl-β-D-thiogalactopyranoside(IPTG),and the recombinant protein of MdbHLH2-MBP was successfully detected by polyacrylamide gel electrophoresis(PAGE).The results provide an important basis for further study of the structure and function of MdbHLH2.

关 键 词：苹果 转录因子MdbHLH2 原核表达 异丙基-β-D-硫代半乳糖苷(IPTG) 聚丙烯酰胺凝胶电泳(PAGE) 

分 类 号：Q943.2[生物学—植物学]