Construction of a Normalized cDNA Library of Barley Embryo Treated with Exogenous ABA

It is necessary to construct the cDNA library of barley embryo before screening the proteins interacted with target proteins.In the experiment reported in this paper,barley seeds were treated with5μmol·L-1 ABA and distilled water for 0h,6h,12hand 24h,respectively.Barley embryos were stripped and total RNA was extracted.Ds-cDNA was synthesized by RT-PCR with SMART method.The ds-cDNA which digested by Sfi I was ligated to the vectors pGADT7-Rec.All the recombinant vectors were electroporated into DH10Bto construct the cDNA library.The detection results showed that the cDNA library contained 1.1×106 recombinant clones and that all the inserted cDNA fragments were more than 1kb in size,between 1kb and 3kb and most were around 1.2kb,which meets the requirement of the yeast two-hybrid library.