Antibody Production of Baculovirus-expressed VP6 from Porcine Group C Rotavirus

ABSTRACT

The emerging pathogen, group C rotavirus (RVC) has been reported to cause acute diarrhea. But there was the limitation on the detection and monitoring for the absence of rapid sensitive diagnosis system. For the molecular biology study and diagnostic system development, we could detect porcine RVC by reverse transcriptase PCR (RT-PCR) analyses from 60 diarrheal disease porcine stool samples. VP6 full length RT-PCR product (CA-2 RVC, 1352 bp) was cloned and compared the nucleotide and deduced amino acid sequences with those of previously reported other porcine, human, and bovine rotavirus group A, B and C strains. Analyses data showed >82% homology on the nucleotide sequences and >90% homology on the deduced amino acid sequences with other RVCs. Recombinant baculovirus was prepared with cloned PCR product corresponding to VP6 coding sequence (CDS) (position 22~1206) into BaculoDirect(TM) C-term linear DNA, and used for the transfection of insect cells. The polyclonal antibody was produced from mice with purified recombinant VP6 and confirmed with western blot. Both of VP6 antigen and antibody, are useful for the development of rapid diagnostic system against RVC.