Preparation of chitosan-plasmid DNA nanoparticles encoding zona pellucida glycoprotein-3alpha and its expression in mouse.
In the present study, the porcine zona pellucida (ZP)-3alpha eukaryotic expression vector pVAX1-pZP3alpha was constructed by genetic recombinant technology, then the recombinant plasmid was encapsulated in nanoparticles with chitosan, and the imaging of chitosan/pVAX1-pZP3alpha 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-3alpha eukaryotic expression vector pVAX1-pZP3alpha 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-pZP3alpha 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.
Sun
Cai-Jun
CJ
Research Center for Reproductive Immunology, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong 510632, China.
Pan
Shan-Pei
SP
Xie
Qi-Xuan
QX
Xiao
Luan-Juan
LJ
D000818:Animals; D048271:Chitosan; D003001:Cloning, Molecular; D004527:Egg Proteins / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D010957:Plasmids; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D000072216:Zona Pellucida Glycoproteins; D048271:Chitosan