優(yōu)質(zhì)小麥品種鄭麥158不同發(fā)育時(shí)期籽粒的酵母雙雜交cDNA文庫構(gòu)建

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Construction of Yeast Two-Hybrid cDNA Library in Grains at Different Developmental Stages of Zhengmai 158 with High Quality

Wang Shasha'，He Ning1*，Chao Yue'en'，Wang Gang2，Li Yan3， Zhang Yixin4

（1.WheatResearch Institute，HenanAcademyofAgriculturalSciences/HenanKeyLaboratoryf WheatBiology，Zhengzhou40002，

China；2.HenanAcademyofAgricultural Science，Zhengzhou450o2，China；3.Jiaozuo University，Jiaozuo 4540，China; 4. Beijing Municipal Science & Technology Commission， Beijing 1Ooo81，China）

AbstractHigh-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 diffrent developmental stages （7，14， 21，28，35and 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 capacity of primary and secondary cDNA libraries were 8.0×106CFU and 9.4×106CFU ， respectively. The 24 clones were randomly selected forbacteria liquid PCR detection，and the results showed that allthe clones had insert fragments.The length of the insert fragments mainly concentrated between 7OO bp and 2 OOO bp with a recombination rate of 100% ，which were in expected size. In summary，the yeast two-hybrid cDNA library of Zhengmai158constructed in this study had high quality，and could meet the requirements for high-efficient screening of the genes interacting proteins，which could lay a foundation for screening the interacting proteins of vheatquality related genes and analyzing their regulatory mechanisms.

KeywordsZhengmai 158； High-quality wheat; Yeast two-hybrid; cDNA library

酵母雙雜交技術(shù)（YeastTwo-Hybrid，Y2H）是研究植物蛋白質(zhì)與蛋白質(zhì)相互作用的一種常用技術(shù)[1]，目前已被廣泛應(yīng)用于各物種的相關(guān)研究當(dāng)中。（剩余8809字）