南京农业大学学报2017，Vol.40Issue(5)：812-819,8.DOI:10.7685/jnau.201612049
不结球白菜BrCDF1的克隆和蛋白亚细胞定位及其与BrFKF1蛋白的互作验证
Cloning of BrCDF1 gene and protein subcellular localization and identification of its interaction with BrFKF1 in non-heading Chinese cabbage
摘要
Abstract
[Objectives] The paper aims to elucidate the flowering regulatory mechanism of transcription factor CDF1 in the flowering process of Brassica rapa ssp.chinensis and verify if CDFI interacts with the FKF1 protein.[Methods] A full-length sequence of BrCDF1 was isolated by homologous cloning.The subcellular localization vector pEarlyGate101-BrCDF1-YFP,which was constructed by fusion of BrCDF1 and YFP,was transiently expressed in Nicotiana benthamiana leaves by Agrobacterium tumefaciens-mediated method and detected by confocal microscopy.The interaction between BrCDF1 and BrFKF1 in Brassica rapa was studied by yeast two-hybrid and bimolecular fluorescence complementation methods.[Results] The BrCDFI gene contains an open reading frame of 876 bp in length encoding a protein of 292 amino acids.The alignment of Brassica napus,Brassica oleracea var.oleracea and Raphanus sativus showed that the conservatism of BrCDF1 was 91.11％,88.25％ and 84.76％ respectively in the evolutionary process,which was highly conservative comparing with other species.The detection of yellow fluorescence signal by coufocal showed that BrCDF1 was located in nucleus.And there was an interaction between BrCDF1 and BrFKF1.[Conclusions] In this study,BrCDF1 is located in nucleus and interacts with BrFKF1.BrCDF1 is highly conserved during evolution.BrCDF1 possibly plays a role in photoperiod pathway of flowering in non-heading Chinese cabbage.关键词
不结球白菜/BrCDF1/亚细胞定位/酵母双杂交/双分子荧光互补Key words
Brassica rapa ssp.chinensis/BrCDF1/subcellular localization/yeast two hybrid/bimolecular fluorescence complementation分类
农业科技引用本文复制引用
吴小婷,邵帅旭,李英,张昌伟,侯喜林,沈振国,刘同坤..不结球白菜BrCDF1的克隆和蛋白亚细胞定位及其与BrFKF1蛋白的互作验证[J].南京农业大学学报,2017,40(5):812-819,8.基金项目
国家自然科学基金项目(31301782) （31301782）
中央高校基本业务费专项资金(Y0201700179) （Y0201700179）
江苏省自然科学基金项目(BK20171374) （BK20171374）
中国博士后科学基金项目(2014M550294,2015T80561) （2014M550294,2015T80561）
