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白桦开花抑制因子BpFLC与BpSVP酵母表达载体的构建及互作验证 被引量：2

Determination of Interactions between BpSVP and BpFLC from Betual platyphylla Suk. by Yeast Two-hybrid System
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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,利用PCR技术从白桦幼叶cDNA中克隆出BpFLC和BpSVP基因序列,利用NdeⅠ和EcoRⅠ限制性内切酶将BpFLC和BpSVP分别整合进酵母表达载体pGBKT7和pGADT7中。pGBKT7-BpFLC及pGBKT7-BpSVP转化酵母Y2Hgold菌株显示无自激活与毒性作用。酵母Y2Hgold[pGBKT7-BpFLC×pGADT7-BpSVP]、Y2Hgold[pGADT7-BpFLC×pGBKT7-BpSVP]均可在QDO/A/X平板上生长,并显示蓝色,说明能激活报告基因HIS3、ADE2、AUR1-C、MEL1,由此表明BpFLC与BpSVP蛋白能够结合,存在互作关系。 We studied the mechanism of interaction between SVP and FLC in Betual platyphylla. The coding sequences of BpFLC and BpSVP were respectively amplified via PCR from Betual platyphylla young leaves cDNA. The target genes were digested with NdeⅠ and EcoRⅠ,then respectively ligased into yeast expression vector pG BKT7 and pGADT7. After pG BKT7-BpFLC or pGBKT7-BpSVP was transformed into Y2 Hgold yeast strain,there was no self-activation and toxicity. The yeast strains Y2 Hgold [pGBKT7-BpFLC×pG ADT7-BpSVP] and Y2 Hgold [pGADT7-BpFLC × pGBKT7-BpS VP] grew on selective agar plates QDO / A / X with blue stains. BpSVP and BpF LC could combine with each other,form a complex,and activate transcription of four independent reporter genes HIS3,ADE2,AUR1-C,MEL1. There was a direct interaction between BpFLC and BpSVP.
作者 郑唐春 臧丽娜 代丽娟 刘彩霞 曲冠证
机构地区 林木遗传育种国家重点实验室(东北林业大学)
出处 《东北林业大学学报》 CAS CSCD 北大核心 2015年第8期64-70,共7页 Journal of Northeast Forestry University
基金 国家高技术研究发展计划项目(2011AA100202-1-6)资助
关键词 白桦 BpFLC基因 BpSVP基因 酵母双杂交 蛋白互作 Betual platyphylla BpFLC BpSVP Yeast two-hybrid Protein interaction
分类号 Q784 [生物学—分子生物学] Q939.5 [生物学—微生物学]
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参考文献29

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东北林业大学学报
2015年 第8期

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