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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Vu MYB1。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c DNA文库,以Vu MYB1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Vu MYB1互作的蛋白。结果表明:c DNA文库库容为1.25×107CFU,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Vu MYB1互作的蛋白。该研究有助于进一步解析Vu MYB1调控花青苷生物合成的分子机理。
Abstract：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 Vu MYB1 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 c DNA library.The bait vector was constructed with Vu MYB1 as the bait protein, and the yeast two-hybrid library was screened through the yeast two-hybrid system to determine the proteins interacting with Vu MYB1.The results showed that the c DNA library had a capacity of 1.25×107 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 Vu MYB1 were obtained.This study is helpful for further understanding the molecular mechanism of Vu MYB1 in regulating anthocyanin biosynthesis.
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基本信息:
DOI：10.13478/j.cnki.jasyu.2025.01.005
中图分类号:S663.9
引用信息:
[1]张东,陈佳禚,刘佳茹等.笃斯越桔酵母双杂文库的构建及Vu MYB1互作蛋白的筛选[J].延边大学农学学报,2025,47(01):32-39.DOI:10.13478/j.cnki.jasyu.2025.01.005.
基金信息:
国家自然科学基金(32160690)