白桦开花抑制因子BpFLC与BpSVP蛋白互作的结构域筛选与互作验证

doi:10.7525/j.issn.1673-5102.2015.06.013

摘要/Abstract

摘要： 为深入研究白桦开花抑制因子FLC与SVP的相互作用的分子机理。从重组质粒pGBKT7-BpFLC、pGBKT7-BpSVP分别克隆出6个BpFLC截短体（BpFLC1~6）和6个BpSVP截短体（BpSVP1~6），分别编码MI、MIK、K、IKC、KC和C域。在酵母Y2HGold菌中，分别共转诱饵质粒pGBKT7-BpFLC1~6×pGADT7-BpSVP及pGBKT7-BpFLC×pGADT7-BpSVP1~6。酵母转化子Y2HGold［pGBKT7-BpFLC2~5×pGADT7-BpSVP］，可在选择性固体培养基TDO、QTO/A上生长，并在QDO/A/X上长出蓝色菌落，表明BpSVP能与截短体蛋白BpFLC2~5异源结合。此外酵母Y2HGold［pGBKT7-BpFLC×pGADT7-BpSVP2~5］也能同时激活报告基因AUR1-C、HIS3、ADE2、MEL1。进一步研究发现：Y2HGold［pGBKT7-BpFLC3×pGADT7-BpSVP3］存在相互作用，表明BpFLC的K域与BpSVP的K域能够异源结合，是介导BpFLC与BpVP蛋白互作的关键结构域。

关键词: 白桦, BpFLC, BpSVP, 酵母双杂交, 截短体, 蛋白互作

Abstract: The experiment was conducted to study the mechanism of interaction between SVP and FLC in Betula platyphylla. The truncated genes of BpFLC1-6 and BpSVP1-6 were, respectively, cloned from yeast recombination plasmids pGBKT7-BpFLC and pGBKT7-BpSVP. pGBKT7-BpFLC1-6×pGADT7-BpSVP and pGBKT7-BpFLC×pGADT7-BpSVP1-6 were co-transferred into yeast Y2HGold. The yeast strains of Y2HGold［pGBKT7-BpFLC2-5×pGADT7-BpSVP］ could grew on selective agar plates TDO, QTO/A and QDO/X/A with blue stains. BpSVP truncated forms and BpFLC2-5 protein could act with each other to form heterodimers. Then, Y2HGold［pGBKT7-BpFLC×pGADT7-BpSVP2-5］ were brought into proximity to form protein compounds and activate transcription of four independent reporter genes of AUR1-C, HIS3, ADE2 and MEL1. The interactions between BpFLC3 and BpSVP3 were tested to confirm the acting domains. The yeast Y2HGold［pGBKT7-BpFLC3×pGADT7-BpSVP3］ exhibited blue stains on selective agar plates QDO/X/A. The K domain of BpFLC and that of BpSVP were the key structure domains and mediated the protein interactions between BpFLC and BpSVP.

Key words: Betula platyphylla, BpFLC, BpSVP, yeast two-hybrid, truncated form, protein interaction

中图分类号:

臧丽娜；郑唐春；代丽娟；刘彩霞；曲冠证*. 白桦开花抑制因子BpFLC与BpSVP蛋白互作的结构域筛选与互作验证[J]. 植物研究, 2015, 35(6): 866-872.

ZANG Li-Na；ZHENG Tang-Chun；DAI Li-Juan；LIU Cai-Xia；QU Guan-Zheng*. Identification of Protein Interactions between Flowering Repressors BpFLC and BpSVP from Betula platyphylla[J]. Bulletin of Botanical Research, 2015, 35(6): 866-872.

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