A potential approach to antifertility vaccine by expression of sperm membrane peptide in Salmonella.
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, antigenically determinant region IV of Salmonella flagellin gene fliC (d). The recombinant plasmid (pLS408-H1) was transferred into the nonpathogenic 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 immunogold-labeling electron microscopy using mouse anti-YWK-II antiserum. A 60 kD protein, corresponding to the chimeric flagellin, was isolated from the recombinant S. 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-H1 may thus be a potential source of antifertility vaccine.
Gao
A W
AW
Shanghai Institute of Cell Biology, Academia Sinica, Shanghai 200031, China.
Yan
Y C
YC
Yang
Q S
QS
Zhao
F
F
Dong
Q
Q
Zhang
M L
ML
Koide
S S
SS
D016564:Amyloid beta-Protein Precursor; D004926:Escherichia coli / Q000235:genetics; D005407:Flagella / Q000235:genetics; D005408:Flagellin / Q000235:genetics*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D009419:Nerve Tissue Proteins; D009841:Oligonucleotides / Q000235:genetics; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D012475:Salmonella / Q000235:genetics* / Q000378:metabolism; D022642:Vaccines, Contraceptive
C489697:APLP1 protein, human; C084104:APLP2 protein, human; D016564:Amyloid beta-Protein Precursor; C492952:Aplp2 protein, mouse; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D009841:Oligonucleotides; D011994:Recombinant Proteins; D022642:Vaccines, Contraceptive; D005408:Flagellin