By using the SMART technique, a yeast twohybrid cDNA library of rice roots was constructed and utilized to study proteinprotein interactions induced by drought. An optimized high efficiency membrane mating method, substituting for liquid mating method, was developed for library screenings. As results, there are about 4.9 ×106 clones in the library with the averaged size of the insert fragments to be about 800 base pairs. To verify the efficiency of the membrane mating method, OsWRKY71, a wellstudied gene encoding a transcriptional repressor of gibberellin signaling, was used as a bait to screen this library. The mating efficiency was shown to be as high as 6.9%, which was six times higher than that of the liquid mating method. This high efficient mating method is simple, costeffective and suitable for large scale yeast hybrid analyses.
FU Jian1, Linkun GU 2, GUO Yiqing1, Liyuan ZHANG 2, WANG Linxian1, LI Dingqin1, WANG Bo1, Jeff Qingxi SHEN2,*, CHENG Zaiquan1,*
. Construction and Highefficient Screenings of a Yeast TwoHybrid cDNA Library from the Droughtstressed Roots of Rice[J]. Chinese Journal OF Rice Science, 2013
, 27(2)
: 198
-202
.
DOI: 10.3969/j.issn.10017216.2013.02.013
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