Construction of Yeast Two-Hybrid cDNA Library in Grains at Different Developmental Stages of Zhengmai 158 with High Quality
Wang Shasha
He Ning
Chao Yue'en
Wang Gang
Li Yan
Zhang Yixin
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.
Keywords：Zhengmai 158High-quality wheatYeast two-hybridcDNA library
Publication Date：2025-04-30
Online Publishing Date：2026-05-22(First online date of this platform, not the publication date of the document)
Pages：6( 40-45 )