大叶落地生根(Kalanchoe daigremontiana)KdSOC1基因的克隆及自激活检测  

Cloning and Self-activation Detection of KdSOC1 Gene from Kalanchoe daigremontiana

作　　者：王梦迪 许立新[1] 董笛 赵方东 晁跃辉[1] 曾会明[1] Wang Mengdi;Xu Lixin;Dong Di;Zhao Fangdong;Chao Yuehui;Zeng Huiming(Turfgrass Research Institute of Beijing Forestry University, Beijing, 100083)

机构地区：[1]北京林业大学草坪研究所

出　　处：《分子植物育种》2019年第13期4267-4273,共7页Molecular Plant Breeding

基　　金：北京林业大学科技创新计划项目(2018ZY35)资助

摘　　要：大叶落地生根(Kalanchoe daigremontiana)为景天科多年生肉质草本植物,是研究无性繁殖的模式植物。SOC1 (Suppressor of overexpression of constans 1)基因是植物成花过程中MADS-box基因家族中的关键转录因子。为深入研究大叶落地生根KdSOC1基因的功能,于大叶落地生根中克隆KdSOC1基因,进行生物信息学分析,构建酵母双杂交诱饵表达载体pGBKT7-KdSOC1,并采用LiAc法将重组质粒pGBKT7-KdSOC1转化至Y2HGold酵母菌株中,通过营养缺陷型培养基培养进行自激活检测。结果表明,成功克隆了KdSOC1基因,该基因开放阅读框共有684 bp。生物信息学分析结果显示,Kd SOC1蛋白共有227个氨基酸残基,分子量为25.88 kD,大叶落地生根SOC1蛋白与牡丹的亲缘关系最为接近。双杂交诱饵表达载体pGBKT7-KdSOC1构建成功,并在酵母细胞中正常表达,表达产物对报告基因无自激活作用。本研究为筛选大叶落地生根中与其互作的蛋白及后续实验提供了帮助。Kalanchoe daigremontiana is a perennial succulent herb of sedum, which is a model plant for the study of asexual reproduction. The SOC1(Suppressor of overexpression of constans 1) gene is a key transcription factor in the MADS-box gene family during plant flowering. In order to further study the function of KdSOC1 of Kalanchoe daigremontiana, KdSOC1 gene was cloned from Kalanchoe daigremontiana for bioinformatics analysis, and yeast two-hybrid bait expression vector pGBKT7-KdSOC1 was constructed. Recombinant plasmid pGBKT7-KdSOC1 was transformed into Y2 HGold yeast strain by Li Ac method, and self-activation assays were performed by auxotrophic medium culture. The results showed that the KdSOC1 gene was successfully cloned, and the open reading frame of the gene was 684 bp. Bioinformatics analysis showed that KdSOC1 protein had 227 amino acid residues and the molecular weight was 25.88 kD. The SOC1 protein of Kalanchoe daigremontiana had the highest homologous characteristics with peony. The two-hybrid bait expression vector pGBKT7-KdSOC1 was successfully constructed and expressed in yeast cells. The expression product had no self-activation effect on the reporter gene.This study would be helpful for screening proteins interacting in the Kalanchoe daigremontiana and conducting subsequent experiments.

关 键 词：大叶落地生根 KdSOC1基因 酵母双杂交 诱饵表达载体 自激活检测 

分 类 号：Q943.2[生物学—植物学]