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Online Journal of Veterinary Research©
Volume 26 (7): 567-581, 2022.
Immuno clone for Streptococus agalactiae FbsA DNA mastitis vaccine.
Mohsen Amini1, Azam Mokhtari*1,2, Ali Kadivar3, Somayeh Shahrokh1
1Department of Pathobiology, 2 Zoonotic Disease 3Clinical Sciences Faculty of Veterinary Medicine, Research Institute, Shahrekord University, Shahrekord, Iran
Amini M, Mokhtari A, Kadivar A, Shahrokh S., Immuno clone for Streptococus agalactiae FbsA DNA mastitis vaccine, Onl J Vet Res., 26 (7): 567-581, 2022. FbsA virulence factor in Streptococcus agalactiae induces mastitis in dairy cows. We cloned FbsA gene from Streptococcus agalactiae for immunization against mastitis. Sequence of conserved region of gene was extracted from NCBI database and immunogenicity of protein was evaluated. After alignment, ring-infected erythrocyte surface antigen domain amino acids 89 to 292 were selected with cleavage sites at BamH I ´5 and EcoR I ´3 expressed without mismatch. Twelve amino acid epitopes within protein could generate ~ 10 antibodies for humoral immunity. We predicted cell immunogenicity finding ~25 epitopes recognized by B lymphocytes with 9 amino acids for antigen binding. The desired fragment was cloned into the plasmid pCDH-CMV-MCS-EF1-eGFP-T2A-Puro confirming recombinant plasmid and cloning by restriction enzymes, colony PCR and sequencing.
Keywords: Mastitis, Streptococcus agalactiae, FbsA, DNA.