苹果响应钙信号转录因子 MdbHLH2 原核表达分析

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 MdbHLH2 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- $ \beta $-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 MdbHLH2 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.