Fig. 1

Production of recombinant parasites. a Confirmation of integrating pLEXSY-neo2.1/LACK-KMP11-EGFP and pLEXSY-neo2.1/EGFP into the parasite genome by PCR. PCR was performed on the genome of transfectants that show the 4295-bp band (Lanes 1, 2) representative of this gene in the L. tarentolae-LACK/KMP11/EGFP. PCR was performed on the genome of the wild parasite, which does not show a band (negative control) (Lanes 3, 4). b PCR was carried out on the genome of L. tarentolae- EGFP, which shows the 918-bp band representative of this gene in the L. tarentolae- EGFP strain (Lanes 3, 4). PCR was carried out on the genome of the wild parasite, which does not show a band (control) (Lanes 1, 2). c Confirmation of the LACK protein by Western blot analysis. Lanes 2, 3 produced 38 kDa bands from the LACK protein extracted from the culture medium of recombinant L. tarentolae-LACK/KMP11/EGFP parasite using Anti-Hig-tag antibody. d Western blot analysis conducted to confirm KMP11 protein. Lanes 2, 3 produced 13 kDa bands from the KMP11 protein extracted from the culture medium of recombinant L. tarentolae-LACK/KMP11/EGFP parasite using Anti-S-tag antibody. e The recombinant live vaccine of L. tarentolae-LACK/KMP11/EGFP containing pLEXSY-neo2.1/LACK-KMP11-EGFP constructs observed by fluorescent microscopy and (f) L. tarentolae-EGFP promastigotes