A POTENTIAL APPROACH TO ANTIFERTILITY VACCINE BY EXPRESSION OF SPERM MEMBRANE PEPTIDE IN SALMONELLA
GAO Ai Wu YAN Yuan chang YANG Qi Sheng ZHAO Feng DONG Qi ZHANG Mei Lan S. S.KOIDE (Shanghai Institute of Cell Biology, Academia Sinica,Shanghai 200031 ,China) (Biology Department of Zhejiang University, Hangzhou 310027.China) Center for Biomedical Research,The Population Council, 1230 York Avenue,New York, NY, 10021,USA)
A synthetic oligonucleotide, HSD-2a , encoding a peptide segment of the extracellular domain of a human sperm membrane protein, YWK-Ⅱ, was inserted by blunt-end ligation at the EcoRV site in the hypervariable, antigeni-cally determinant region IV of Salmonella flag-ellin gene fliC(d). The recombinant plasmid (pLS408-H1) was transferred into the non-pathogenic aroA live vaccine Salmonella dublin strain SL5928, which is flagellin-negative. Location of the HSD-2a protein on the surface of the flagella was demonstrated by ELISA and by im-munogold-labeling electron microscopy using mouse anti-YWK- Ⅱ antiserum. A 60 kD protein, corresponding to the chimeric flagellin, was isolated from the recombinant 5. dublin and found to contain the expressed HSD-2a peptide determined by Western blot analysis using anti-YWK- Ⅱ antiserum. The Salmonella strain carrying the recombinant plasmid-pLS408-Hl may thus be a potential source of antifertility vaccine.