›› 2011, Vol. 54 ›› Issue (11): 1324-1328.doi:

• SHORT COMMUNICATIONS • Previous Articles     Next Articles

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

 LI  Shuo, SUN  Li-Juan, LI  Xing, XIONG  Ru-Yi, XU  Qiu-Fang, ZHOU  Yi-Jun   

  • Received:2011-07-29 Revised:2011-09-25 Online:2011-11-20 Published:2011-11-20
  • Contact: ZHOU Yi-Jun E-mail:yjzhou@jaas.ac.cn
  • About author:lishuolstonnv989@163.com;2009102016@njau.edu.cn

Abstract:  In order to research the interaction between the small brown planthopper (Laodelphax striatellus Fallén, 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.

Key words: Small brown planthopper (Laodelphax striatellus), rice stripe virus, homologous recombination, cDNA library, yeast two-hybrid cDNA library