‘秦冠’苹果MdWRKY基因亚细胞定位及原核表达

Studies on Subcellular Localization and Expression in E.coli of MdWRKY from‘Qinguan’Apple

以‘秦冠’苹果为试材,采用RT-PCR方法获得了一个WRKY基因,全长1 224 bp,推断其编码331个氨基酸,命名为MdWRKY。亚细胞定位分析MdWRKY蛋白分布在细胞核内,属于核蛋白。实时定量分析结果显示,MdWRKY基因受苹果斑点落叶病菌的诱导,表明其可能参与植物与病原菌的互作。随后进行原核表达分析,SDS/PAGE电泳结果表明表达蛋白与其蛋白大小一致。

A WRKY gene,designated as MdWRKY,was isolated from‘Qinguan’apple leaves using reverse transcription PCR approaches.MdWRKY was 1 224 bp and encoded a 331 amino-acid protein.Analysis of subcellular localization indicated that MdWRKY protein was targeted to the nucleus.Quantitative RT-PCR analysis showed that MdWRKY transcription was induced by Alternaria alternata f sp.mali indicating that MdWRKY gene may be involved in the interaction between plant and pathogen.Then expression in E.coli was conducted,SDS/PAGE result demonstrated that expression proteins consistent with the size of expected protein,which provided a foundation for further purifying and function study of MdWRKY.