Abstract: In order to explore the interactions between grass carp reovirus and the host cell proteins, the yeast two-hybrid cDNA library from grass carp CIK（Ctenopharyngodon idellus kidney） was constructed by SMART technology. Total RNA of the CIK cells was extracted and mRNA was purified. Then, mRNA as the template, the first strand of cDNA was synthesized by reverse transcription. The double-stranded cDNA was amplified through the long-distance PCR with the DNA polymerase. The cDNA library from grass carp CIK cells was constructed in the yeast strain 187, using SMART technology and the homologous recombination method. After testing, the conversion rate and the capacity of the original library was 1.6×10⁵ and 2.4×10⁶, respectively. The length of the inserted double-stranded cDNA fragments were 250-2 000 bp. The titer of the library was 7×10⁷ CFU/mL and the recombination rate was 98%. This library had the well polymorphism and the integrity of the cDNA fragments. Using the constructed yeast two-hybrid from the CIK cells, the VP7 and VP5 protein in grass carp reovirus were as the baits for the screening experiment. The positive colonies of the interacting proteins of VP7 had been acquired, without the positive colony of the interacting protein of VP5. The construction of the yeast two-hybrid cDNA library from the grass carp CIK cells provided an important research tool for the study on the interaction mechanism of grass carp reovirus with the host cell.

Key words: Ctenopharyngodon idellus kidney（CIK）, Yeast two-hybrid cDNA library, Protein interaction