Screening of Host Proteins Interacting with Pc2 of Rice Stripe Virus by Yeast Two-hybrid System
Rice stripe virus encoded Pc2 protein is involved in the viral infection and symptom formation of rice.Study on the interaction between Pc2 and host is essential to elucidate the molecular mechanism of viral infection.In this study,the encoding region of Pc2-N or Pc2-C was fused to the pGBKT7,the rice cDNA library was constructed and screened by the Pc2-N or Pc2-C as a bait to identify proteins that interacted with Pc2.The results showed that the coding regions and the amino acid sequences of insert Pc2-N and Pc2-C were correct.Pc2-N had neither toxicity nor self activating effect on AH109,while Pc2-C was toxic to AH109.The cDNA library had a titer o f 4×107 CFU/mL,and most of insert fragments were between 500 bp to 2 000 bp in size.Eight candidate positive clones were obtained by screening with Pc2-N as a bait.Their cDNA Sequences were obtained and BLAST results showed that these cDNAs encoded 5 proteins,which mainly were the proteins involved in photosystem and stress.