山东农业科学2025，Vol.57Issue(4)：40-45,6.DOI:10.14083/j.issn.1001-4942.2025.04.005
优质小麦品种郑麦158不同发育时期籽粒的酵母双杂交cDNA文库构建
Construction of Yeast Two-Hybrid cDNA Library in Grains at Different Developmental Stages of Zhengmai 158 with High Quality
摘要
Abstract
High-quality yeast two-hybrid library is the basis for screening the genes interacting proteins and analyzing the regulation mechanisms.In this study,the grains at different developmental stages(7,14,21,28,35 and 42 days after anthesis)of Zhengmai 158 with high quality were equally mixed to construct the yeast two-hybrid cDNA library by gateway recombination technology.The results showed that the library capac-ity of primary and secondary cDNA libraries were 8.0×106 CFU and 9.4×106 CFU,respectively.The 24 clones were randomly selected for bacteria liquid PCR detection,and the results showed that all the clones had insert fragments.The length of the insert fragments mainly concentrated between 700 bp and 2 000 bp with a recom-bination rate of 100％,which were in expected size.In summary,the yeast two-hybrid cDNA library of Zheng-mai 158 constructed in this study had high quality,and could meet the requirements for high-efficient screen-ing of the genes interacting proteins,which could lay a foundation for screening the interacting proteins of wheat quality related genes and analyzing their regulatory mechanisms.关键词
郑麦158/优质小麦/酵母双杂交/cDNA文库Key words
Zhengmai 158/High-quality wheat/Yeast two-hybrid/cDNA library分类
农业科学引用本文复制引用
王沙沙,何宁,晁岳恩,王刚,李艳,张伊欣..优质小麦品种郑麦158不同发育时期籽粒的酵母双杂交cDNA文库构建[J].山东农业科学,2025,57(4):40-45,6.基金项目
河南省科技攻关项目(232102110220) （232102110220）
河南省农业科学院自主创新项目(2024ZC001) （2024ZC001）
