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Expression of recombinant mouse sperm protein sp56 and assessment of its potential for use as an antigen in an immunocontraceptive vaccine
Author(s) -
Hardy Christopher M.,
Mobbs Karen J.
Publication year - 1999
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/(sici)1098-2795(199902)52:2<216::aid-mrd13>3.0.co;2-r
Subject(s) - biology , polyclonal antibodies , recombinant dna , antiserum , fusion protein , microbiology and biotechnology , acrosome reaction , sperm , antigen , complementary dna , acrosome , epitope , adjuvant , virology , immunology , gene , biochemistry , genetics
Recombinant mouse sp56 protein was produced for testing as an antigen in an immunocontraceptive vaccine. The coding sequence for the mature sp56 protein was cloned into the bacterial expression system pFLAG using a PCR‐based method on mouse testis cDNA. Polyclonal antisera were raised in mice against affinity purified recombinant sp56 fusion protein (sp56FLAG) or an artificial sp56 peptide fused to a carrier protein (KLH) and shown to cross‐react to a protein band of 75 kD in detergent extracts of mouse sperm by Western immunoblot analysis under reducing conditions. The antisera to sp56FLAG also immunolocalized over the entire acrosome of mouse sperm. Female BALB/c mice were immunized intraperitoneally with sp56FLAG in a fertility trial with 20 μg sp56FLAG in Freund's Complete Adjuvant and boosted three to five times with 20 μg sp56FLAG in Freund's Incomplete Adjuvant. Litter sizes of sp56FLAG‐treated mice were significantly smaller than control‐treated animals after five boosts. Mol. Reprod. Dev. 52:216–224, 1999. © 1999 Wiley‐Liss, Inc.