Production and purification of recombinant non toxin soluble antigen sag a protein from equine streptococci

Production and Purification of Recombinant Non Toxin Soluble Antigen Sag A Protein from Equine Streptococci

Using ex vivo models of equine respiratory tract tissue has shown that SLS contributes to the ability of the equine streptococci to colonies and damage equine respiratory tract tissue. This suggests that if it were possible to elicit neutralizing anti-SLS antibodies in horses, these could contribute to protection against the equine streptococcal diseases. For this purpose recombinant GST-sagA encoded plasmid have been constructed by fusion of 166 nucleotide of sagA gene (which obtained by PfʹR) from streptococcus equi to pGEX-3X plasmid. Recombinant plasmid was transformed to three different competent Ecoli B121, Co+ and Plyses cells. Recombinant GST-SagA was induced by IPTG (] n1M -1M) for 4hr and purified by FFI column. It was shown that BL21 produced more recombinant protein and highly purified recombinant GST- SagA protein were obtained by FFI column. This soluble peptide was desalted and concentrated with viva science filter tube and 100 ml of 10 mgml of this peptide was obtained from 2000ml cell culture.

Using ex vivo models of equine respiratory tract tissue has shown that SLS contributes to the ability of the equine streptococci to colonies and damage equine respiratory tract tissue. This suggests that if it were possible to elicit neutralizing anti-SLS antibodies in horses, these could contribute to protection against the equine streptococcal diseases. For this purpose recombinant GST-sagA encoded plasmid have been constructed by fusion of 166 nucleotide of sagA gene (which obtained by PCR) from streptococcus equi to pGEX-3X plasmid. Recombinant plasmid was transformed to three different competent Ecoli B121, Co+ and Plyses cells. Recombinant GST-SagA was induced by IPTG (1 mM -1M) for 4hr and purified by FF1 column. It was shown that BL21 produced more recombinant protein and highly purified recombinant GST- SagA protein were obtained by FF1 column. This soluble peptide was desalted and concentrated with viva science filter tube and 100 ml of 10 mg\ml of this peptide was obtained from 2000ml cell culture.