Construction of yeast two-hybrid cDNA library of high-viruliferous(RSV-infected) populations of the small brown planthopper,Laodelphax striatellus(Hemiptera:Delphacidae)

In order to research the interaction between the small brown planthopper(Laodelphax striatellus Fallen,SBPH) and rice stripe virus(RSV),yeast two-hybrid cDNA library of high-viruliferous(RSV-infected) SBPH populations was constructed.SBPH infected with RSV was maintained in our laboratory.The mRNA was isolated and purified from high-viruliferous populations,and used as the template to synthesize the double-strand cDNAs.The cDNAs were ligated with three-frame adapter and purified by the cDNA size fractionation columns.By using homologous recombination reaction,the cDNA entry library was prepared and then recombined into Gateway compatible vector pGADT7-DEST to construct yeast two-hybrid cDNA library.Detection of the library indicated that it contained 3.68×107 independent clones,and the titer of the amplified library was 2.62×1010 cfu/mL.The recombination rate was above 95%,and the average size of inserts was above 1 kb in the cDNA library.The results demonstrate that the library database meets the requirements of the standard cDNA library.The yeast two-hybrid cDNA library of high-viruliferous SBPH will be useful for the future research on the interaction between insect vector and RSV.