CAJ | 학술논문

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cD-NA中克隆出BpFLC和BpSVP基因序列，利用NdeⅠ和Eoc RⅠ限制性内切酶将BpFLC和BpSV P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－B pSVP]、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BpS-VP蛋白能够结合，存在互作关系。
위심입연구백화개화억제인자FLC여SVP적상호작용적분자궤리，이용PCR기술종백화유협cD-NA중극륭출BpFLC화BpSVP기인서렬，이용NdeⅠ화Eoc RⅠ한제성내절매장BpFLC화BpSV P분별정합진효모표체재체pGBKT7화pGADT7중。 pGBKT7－BpFLC급pGBKT7－BpSVP전화효모Y2Hgold균주현시무자격활여독성작용。효모Y2Hgold [ pGBKT7－BpFLC ×pGADT7－B pSVP]、Y2Hgold [ pGADT7－BpFLC ×pGBKT7－BpSVP]균가재QDO／A／X평판상생장，병현시람색，설명능격활보고기인HIS3、ADE2、AUR1－C 、MEL1，유차표명BpFLC여BpS-VP단백능구결합，존재호작관계。
We studied the mechanism of interaction between SVP and FLC in Betual platyphylla.The coding sequences ofB pFLC and BpSVP were respectively amplified via PCR from Betual platyphylla young leaves cDNA.The target genes were digested with NdeⅠand EcoRⅠ, then respectively ligased into yeast expression vector pGBKT7 and pGADT7.After pGBKT7-Bp-FLC or pGBKT7-BpSVP was transformed into Y2Hgold yeast strain, there was no self-activation and toxicity.The yeast strains Y2Hgold [pGBKT7-BpFLC×pGADT7-BpSVP] and Y2Hgold [pGADT7-BpFLC×pGBKT7-BpSVP] grew on selec-tive agar plates QDO/A/X with blue stains.BpSVP and BpFLC could combine with each other, form a complex, and acti-vate transcription of four independent reporter genes HIS3, ADE2, AUR1-C, MEL1.There was a direct interaction between BpFLC and BpSVP .