小鼠卵透明带3基因疫苗体外瞬时表达的研究

The Construction of Murine ZP3 DNA Vaccine and Its Transient Expression in HeLa Cell

运用基因重组技术 ,将小鼠 ZP3c DNA片段克隆到真核表达载体 p CDNA3上 ,成功构建了 p CDNA3- ZP3真核表达质粒 ,转染 He L a细胞后 ,EL ISA法检测到 p CDNA3- ZP3真核表达质粒在 He L a细胞瞬时表达系统中能够特异性表达ZP3蛋白 ,结果表明 p CDNA3-

The Zona pellucida 3(ZP3) protein, serves as primary sperm receptor,plays an important role in the process of fertilization. Since its antibody against murine ZP3 can effectively inhibit fertilization in mouse, murine ZP3 has become a promising candidate immunogen for controlling the pest mice fertility. The eukaryotic expression vector pCDNA3 was chosen as a vector to construct the DNA vaccine-pCDNA3 ZP3 and then the recombinant plasmid pCDNA3 ZP3 was transformed into E.coli DH5α to be screened for the positive clones. The purified pCDNA3 ZP3 plasmid was transfected into HeLa cells to produce specific murine ZP3 protein. ELISA with anti ZP3 antibody showed that the HeLa cells transfected with pCDNA3 ZP3 can produce ZP3 protein. The result indicated that the recombinant plasmid pCDNA3 ZP3 could be considered as a contraceptive vaccine for the future immunization research on mouse.