Mr. Mott et W. Reilly, DOUBLE IMMUNOGOLD LABELING DEMONSTRATING EXPRESSION OF RECOMBINANT GENES FOR PRODUCTION OF AN ANTI-FERTILITY VACCINE, Micron, 25(6), 1994, pp. 539-545
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.