Preparation of chitosan‐plasmid DNA nanoparticles encoding zona pellucida glycoprotein‐3α and its expression in mouse

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Abstract
In the present study, the porcine zona pellucida (ZP)-3α eukaryotic expression vector pVAX1-pZP3α was constructed by genetic recombinant technology, then the recombinant plasmid was encapsulated in nanoparticles with chitosan, and the imaging of chitosan/pVAX1-pZP3α nanoparticles by Atomic Force Microscope (AFM) was processed. Feeding mouse with those microencapsulation by gastric larvae, and after 5 days, detecting its expression in mouse intestine by RT-PCR and indirect immunofluorescence (IIF). Results show that the porcine ZP-3α eukaryotic expression vector pVAX1-pZP3α had been constructed correctly, and the chitosan-DNA expressing ZP microencapsulation was prepared successfully. After 5 days of feeding mouse, the transcription and expression of those DNA vaccines were found in mouse alvine chorion. The preparation of chitosan/pVAX1-pZP3α plasmid DNA nanoparticles and its expression in mice will help to investigate the feasibility of ZP DNA vaccine to induce oviduct local mucosal immunity against ZP to block the fertilization without causing ovarian dysfunction, which will provide new ideas and ways for research and exploiting more effective, more convenient oral contraceptive vaccines. Mol. Reprod. Dev. 68: 182–188, 2004.