Fig. 1.

The constitution of recombinant B. subtilis-gCa and B. subtilis-gDa. a The recombinant B. subtilis-gCa and B. subtilis-gDa were constructed by common molecular biology technique. The PRV gCa, gDa, EGFP gene were connected by overlap PCR, then the specific connected fragments were inserted into the linearized plasmid p43NMK by in-fusion clone to get p43NMK-gCa-EGFP and p43NMK-gDa-EGFP. Furthermore, the two recombinant plasmids were transformed into B. subtilis WB800 by electroporation. The fusion proteins of B. subtilis-gDa (b) and B. subtilis-gCa (c) were detected by western blotting in 24 h, 48 h and 60 h. Besides, the green fluorescence were observed the B. subtilis-gCa (e) and B. subtilis-gDa (d) by confocal microscope. f The schematic of the immunization, the intranasal administration was performed at 0 and 14 day. In addition, the black triangle (under the line) indicated the time point of sampling the washings (including BAL and vagina) and serum, the red triangle (under the line) represented the last sampling and sampled the washings, serum and spleen