DOUBLE IMMUNOGOLD LABELING DEMONSTRATING EXPRESSION OF RECOMBINANT GENES FOR PRODUCTION OF AN ANTI-FERTILITY VACCINE

The multiple antibody technique for double immunogold labelling for th e simultaneous localization of two antigens with negative staining was utilized to demonstrate the expression of recombinant genes in bacter ia. with the primary antibodies being raised in different host species . For the production of a vaccine for immunological control of fertili ty, a multi-functional plasmid vector was introduced into the bacteriu m Pseudomonas aeruginosa containing the Dichelobacter nodosus fimbrial subunit gene with a grafted amino acid sequence of luteinizing hormon e releasing hormone (LHRH) peptide. Fimbriae of this recombinant, when run on a SDS-polyacrylamide electrophoresis gel, gave a single broad band for LHRH peptide/D, nodosus subunit and were harvested to produce the anti-fertility vaccine which, when injected into mice, produced a trophy of the testes with absence of sperm, resulting in reversible ca stration. Double immunogold labelling of the recombinant P. neruginosa bacteria demonstrated fimbriae with strong expression of the LHRH-pep tide, expression of the D. nodosus subunit and absence of host fimbria e.