As the most powerful and widely used binary interaction assay,the yeast two-hybrid(Y2H)system has been developed for more than 30 years to improve the efficiency and quality of protein-protein interaction(PPI)exploration and genome-wide interactome screening(Fields and Song,1989;Venkatesan et al.,2009;Vidal and Fields,2014).One of the main constraints of high-throughput Y2H screening is the lack of high-quality libraries.Until recently,the protein-encoding open reading frames(ORFeomes)were only available for a few model organisms(Reboul et al.,2003;Gong et al.,2004;Rual et al.,2004;Matsuyama et al.,2006;Yang et al.,2011;Wierbowski et al.,2020).Compared to ORFeomes,the cDNA libraries(including some commercially available cDNA libraries)are a cheap and readily available alternative forY2H screening.However,screening with the cDNA library could result in some false-positives and/or false-negatives in contrast to ORFeomes.False-positive might be caused by the misuse of short ORFs fused to the binding domain,while the false-negative might occur because putative inter-acting proteins are expressed at relatively low levels.Although several efforts have been made to improve the quality of cDNA li-braries(Maier et al.,2011;Yu et al.,2020),more experimental and computational studies are warranted to enhance the efficiency and accuracy of Y2H screening with the cDNA library.

Yinghui Gu;Guannan Li;Ping Wang;Yan Guo;Jingrui Li

State Key Laboratory of Plant Physiology and Biochemistry,College of Biological Sciences,China Agricultural University,Beijing 100193,China

遗传学报

[1]Yinghui Gu;Guannan Li;Ping Wang;Yan Guo;Jingrui Li-.A simple and precise method(Y2H-in-frame-seq)improves yeast two-hybrid screening with cDNA libraries)[J].遗传学报,2022(06):595-598

Venkatesan,ORFeomes,Reboul,Rual,Matsuyama,Wierbowski,forY2H,braries

simple,precise,method,seq,improves,yeast,two,hybrid,screening,cDNA,libraries,most,powerful,widely,binary,interaction,assay,system,has,been,developed,more,than,years,efficiency,quality,PPI,exploration,genome,interactome,Fields,Song,Vidal,One,constraints,high,throughput,lack,Until,recently,encoding,open,reading,frames,were,only,available,few,model,organisms,Gong,Yang,Compared,including,some,commercially,cheap,readily,alternative,However,library,could,result,false,positives,negatives,contrast,False,might,caused,by,misuse,short,ORFs,fused,binding,domain,while,occur,because,putative,acting,proteins,expressed,relatively,low,levels,Although,several,efforts,have,made,Maier,Yu,experimental,computational,studies,warranted,enhance,accuracy

0.481079