MOESM1 of Salmonella Gallinarum delivering M2eCD40L in protein and DNA formats acts as a bivalent vaccine against fowl typhoid and H9N2 infection in chickens

Additional file 1. Analysis of M2eCD40L protein displayed on the surface of SG. M2e gene was physically linked to chicken CD40L peptide and cloned into constitutive pJHL65 expression vector and the recombinant plasmid was subsequently electroporated into attenuated SG mutant strain, JOL2074. The bacterial M2eCD40L surface expression was analysed by flow cytometry using primary M2e-specific polyclonal antibody and then Alexa Fluor 488-conjugated species-specific secondary antibody body. (A) Gating of SG bacteria excluding debris and dead cells. (B) FACS histogram of unstained bacterial control, JOL2074 (B) FACS histogram of stained JOL2068 bacteria lacking M2e gene. (C) FACS histogram of stained JOL2074 bacteria expressing M2e protein showing dramatic increase in fluorescence