烟台大学学报（自然科学与工程版）2025，Vol.38Issue(3)：325-330,6.DOI:10.13951/j.cnki.37-1213/n.240316
大豆酵母双杂交混合文库构建及GmCTP互作蛋白筛选
Construction of Soybean Yeast Two-Hybrid Mixed Library and Screening of GmCTP Interacting Proteins
摘要
Abstract
The Gateway technology was used to construct a yeast two-hybrid library from the root,stem,leaf and flower tissues of the excellent soybean variety Jidou 17.This library was used to screen for interacting proteins of GmCTP encoded by a new gene regulating soybean leaf shape and flower development,providing important basis for the subsequent analysis of the regulatory mechanism of the GmCTP gene.The results showed that the yeast second-ary library had a recombination rate of 100％,a library capacity of CFU=1.44×107,a library titer of 3.6×107 cells/mL,and an average length of the inserted fragment greater than 1000 bp.Self-activation test indicated that the bait vector pGBKT7-CTP had no self-activation activity.Library screening was carried out via yeast mating method,and 110 positive colonies were detected on the four-deficiency medium.Through the four-deficiency and X-α-Gal medium check,45 strong positive blue colonies were obtained.PCR sequencing of positive colonies re-vealed that the function of the LBD family genes in the positive colonies was closely related to the phenotype caused by GmCTP gene mutation,providing important clues for the subsequent molecular mechanism analysis by which the GmCTP gene regulates leaf development.关键词
大豆/酵母双杂交文库/CTP/叶发育Key words
soybean/yeast two-hybrid library/CTP/leaf development分类
农业科技引用本文复制引用
单童,陈磊,徐文静,林越,朱雁,赵婧..大豆酵母双杂交混合文库构建及GmCTP互作蛋白筛选[J].烟台大学学报（自然科学与工程版）,2025,38(3):325-330,6.基金项目
山东省自然科学基金资助项目(ZR2019BC056) （ZR2019BC056）
国家自然科学基金资助项目(32101723). （32101723）
