笃斯越桔酵母双杂文库的构建及VuMYB1互作蛋白的筛选  

Construction of Vaccinium uliginosum yeast hybrid library and screening of VuMYB1 interacting proteins

作　　者：张东 陈佳禚 刘佳茹 魏一 王微 宗成文[1] ZHANG Dong;CHEN Jiazhuo;LIU Jiaru;WEI Yi;WANG Wei;ZONG Chengwen(College of Agriculture,Yanbian University,Yanji Jilin 133002,China;College of Medical,Yanbian University,Yanji Jilin 133002,China)

机构地区：[1]延边大学农学院,吉林延吉133002 [2]延边大学医学院,吉林延吉133002

出　　处：《延边大学农学学报》2025年第1期32-39,共8页Journal of Agricultural Science Yanbian University

基　　金：国家自然科学基金(32160690)。

摘　　要：MYB转录因子在植物中广泛参与次生代谢产物的调控、细胞形态的建成、逆境响应及生长发育等多个方面,同时也是植物花青素生物合成的重要调控因子。该课题组前期筛选到一个可以正向调控花青苷合成的基因VuMYB1。为了深入探究该基因的作用机制,该研究以笃斯越桔蓝果果皮为材料,构建了酵母双杂交cDNA文库,以VuMYB1为诱饵蛋白构建诱饵载体,通过酵母双杂交系统来筛选酵母双杂交文库,确定与VuMYB1互作的蛋白。结果表明:cDNA文库库容为1.25×10^(7)CFU,文库插入片段平均大小约为1.2Kbp,阳性率为100%。通过酵母双杂交筛库及回转验证获得了6种与VuMYB1互作的蛋白。该研究有助于进一步解析VuMYB1调控花青苷生物合成的分子机理。MYB transcription factors are widely involved in the regulation of secondary metabolites,cell morphology formation,stress response and growth and development in plants,and are also important regulatory factors for anthocyanin biosynthesis in plants.In the previous study,a gene VuMYB1 that can positively regulate anthocyanin synthesis was screened.To further explore the mechanism of action of this gene,in this study,the blue fruit peel of Vaccinium uliginosum was used to construct a yeast two-hybrid cDNA library.The bait vector was constructed with VuMYB1 as the bait protein,and the yeast two-hybrid library was screened through the yeast two-hybrid system to determine the proteins interacting with VuMYB1.The results showed that the cDNA library had a capacity of 1.25×10^(7)CFU,the average size of the inserted fragments was approximately 1.2 Kbp,and the positive rate was 100%.Through yeast two-hybrid screening and reverse verification,six proteins interacting with VuMYB1 were obtained.This study is helpful for further understanding the molecular mechanism of VuMYB1 in regulating anthocyanin biosynthesis.

关 键 词：笃斯越桔 VuMYB1 酵母双杂交 蛋白互作 

分 类 号：S663.9[农业科学—果树学]