DEVELOPMENT OF THE RECOMBINANT POSITIVE CONTROLS OF BOVINE VIRAL DIARRHEA VIRUS 1 AND 2 FOR PCR ASSAY

Aim. The method of amplifying and cloning the Erns gene of Bovine Viral Diarrhea Virus was developed to obtain the positive controls for polymerase chain reaction. Methods. The strains used in this study were BVDV-1b (Ossloss) and BVDV-2 (Kosice). Viral RNA was extracted by the silica-based extraction method. Using the specific primers, a part of Erns gene was amplificated. The PCR product was inserted into the cloning vector pTZ57R/T. Furthermore, E. coli DH10B bacteria were transformed to amplify the recombinant plasmid. Recombinant clones were identified by antibiotic selection on agar plate and confirmed by PCR. Moreover, insert of Erns gene was verified by restriction enzyme digestion assay using EcoRI and HindIII. Results. It was shown that we had constructed the recombinant plasmids with insertion Erns gene fragment (826 base pair) of BVDV-1 and BVDV-2. Conclusion. The obtaibed recombinant plasmids can be used as a positive control for PCR.