木薯MeHsfB3b转录因子与MeFKBP20蛋白互作关系验证

Verification of the Interaction Between Cassava MeHsfB3b Transcription Factor and MeFKBP20 Protein

热激转录因子MeHsfB3b是调控木薯抗细菌性枯萎病的重要节点,前期通过酵母双杂交技术获得MeHsfB3b的候选互作蛋白MeFKBP20,该蛋白属于FKBP型肽脯氨酰顺反异构酶基因家族。本研究克隆获得MeFKBP20基因全长为561 bp,编码186个氨基酸,蛋白质分子质量为19.99 kDa,具有FKPB_C结构域。表达模式分析发现:MeFKBP20基因在木薯成熟叶片及根部高表达,在幼叶和叶柄的表达量较低;并能够迅速响应病原菌XpmCHN11诱导,持续保持较高的表达丰度,推测其参与了木薯响应病原菌侵染的过程。利用酵母双杂交点对点、双分子荧光互补实验证明MeHsfB3b蛋白通过在201~241 aa区域与MeFKBP20蛋白作用。本研究结果有助于进一步解析MeHsfB3b基因调控木薯对细菌性枯萎病的抗病机理。

The heat shock transcription factor MeHsfB3b is important in regulating the resistance to cassava bacterial blight.In the previous study,the candidate interaction protein MeFKBP20 of MeHsfB3b was obtained by the yeast two-hybrid technology,which belongs to the FKBP-type peptide prolyl cis-trans isomerase gene family.In this study,the full-length MeFKBP20 gene was cloned to be 561 bp,encoding 186 amino acids,with a molecular weight of 19.99 kDa and a FKPB_C domain.The expression pattern analysis showed that MeFKBP20 was highly expressed in mature leaves and roots of cassava,and the expression level of young leaves and petioles was low.It could quickly respond to the induction of pathogen XpmCHN11 and maintain a high expression abundance,suggesting that it is involved in response to pathogen infection.The interaction between MeHsfB3b protein and MeFKBP20 protein through 201-241 aa region was proved by the yeast two-hybrid point-to-point and bimolecular fluorescence complementation experiments.The results of this study are helpful to further analyze the mechanism of MeHsfB3b gene regulating cassava resistance to cassava bacterial blight.