Prokaryotic expression analysis of calcium signal transcription factor MdbHLH2 in apple

Journal Information

J. Hefei Univ. Tech. (Nat. Sci.), Feb.2025, Vol.48 No.2: 232-235

DOI: 10.3969/j.issn.1003-5060.2025.02.013

Authors

GOU Shasha $ ^{1} $, AN Xin $ ^{1} $, ZHANG Weiwei $ ^{2} $, HU Kangdi $ ^{1} $, ZHANG Hua $ ^{1} $, YAO Gaifang $ ^{1} $

(1. School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China; 2. College of Horticulture Science and Engineering, Shandong Agricultural University, Tai'an 271001, China)

Abstract and Keywords

Abstract: 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.

Keywords: apple; transcription factor MdbHLH2; prokaryotic expression; isopropyl- $ \beta $-D-thiogalactopyranoside(IPTG); polyacrylamide gel electrophoresis(PAGE)

作者信息

苟莎莎 $ ^{1} $，安欣 $ ^{1} $，张玮玮 $ ^{2} $，胡康棣 $ ^{1} $，张华 $ ^{1} $，姚改芳 $ ^{1} $

(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)

摘要和关键词

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 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。

关键词: 苹果;转录因子 MdbHLH2;原核表达;异丙基- $ \beta $-D-硫代半乳糖苷(IPTG);聚丙烯酰胺凝胶电泳(PAGE)

Funding

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)和安徽省自然科学基金资助项目(1908085MC72)

合肥工业大学学报(自然科学版)

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