Production and Purification of Specific IgY Against InvG Protein of Salmonella typhimurium

Salmonella infections are one of the most important causes of human and animal diseases, worldwide. InvG gene is one of the main genes of type III secretion system and leads to initial binding of the bacteria to the host cells. In the recent decades IgY technology has been detected as an efficient alternative procedure to generate antibody for applying in immunotherapy. This research attempted to produce and detect IgY against the recombinant InvG protein of Salmonella typhimurium and evaluate specificity of anti-InvG IgY against the recombinant InvG antigen. Polymerase chain reaction was carried out to amplify InvG gene with InvG one pair of specific primer of Salmonella typhimurium. pTZ57R/T and pET32a vectors were used to clone and sub-clone InvG gene. Recombinant pET32a-InvG plasmid was transferred to E. coli BL21 (DE3). Expression induction was carried out using 1.5 mM isopropyl β-D-1-thiogalactopyranoside. Next, the recombinant InvG protein was injected to laying hens. Western-blotting and Dot-blotting analysis and iELISA were carried out to determine the specify of anti-InvG IgY. This research produced specific InvG egg yolk antibodies by injecting InvG protein to the hens. The anti-InvG IgY corroborated to bind specifically with the InvG protein of Salmonella typhimurium. The finding of the present research confirmed that anti-InvG IgY could be suggested as a candidate for passive immunization and protection against Salmonella typhimurium.