Double immunogold labelling demonstrating expression of recombinant genes for production of an anti-fertility vaccine.
The multiple antibody technique for double immunogold labelling for the simultaneous localization of two antigens with negative staining was utilized to demonstrate the expression of recombinant genes in bacteria, with the primary antibodies being raised in different host species. For the production of a vaccine for immunological control of fertility, a multi-functional plasmid vector was introduced into the bacterium Pseudomonas aeruginosa containing the Dichelobacter nodosus fimbrial subunit gene with a grafted amino acid sequence of luteinizing hormone 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 atrophy of the testes with absence of sperm, resulting in reversible castration. Double immunogold labelling of the recombinant P. aeruginosa bacteria demonstrated fimbriae with strong expression of the LHRH-peptide, expression of the D. nodosus subunit and absence of host fimbriae.
Mott
M R
MR
CSIRO Division of Biomolecular Engineering, New South Wales, Australia.
Reilly
W
W
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D001439:Bacteroides / Q000235:genetics; D003269:Contraception, Immunologic; D010861:Fimbriae, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D015870:Gene Expression; D005798:Genes, Bacterial / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics*; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron / Q000379:methods; D009929:Organ Size; D011550:Pseudomonas aeruginosa / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis*; D013737:Testis; D014611:Vaccination; D014614:Vaccines, Synthetic
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D007987:Gonadotropin-Releasing Hormone