Constyuction Of Myospalax Fontanierii Zona Pellucida 3(MZP3)eukaryotic Systems And Immune Infertility

Chinese zokor(Myospalax fontanierii) belong hamster Branch, is a subfamily zokor, which is the dominant species of rodents of dryland and quantity distribution. Chinese zokor poses a serious threat forestry, animal husbandry, development of agriculture in our country and cause huge economic losses. Current control methods are mainly hunted, physical isolation, drug avoidance, and other toxic agents, while in the control population and reduce the number of trees zokor victims has played a role, but it is facing a heavy workload, the effect is obvious, poor specificity, population recovery, high economic costs and environmental safety, and other issues exist. Therefore, to meet the urgent need for a safe and economical, loyal and strong, new methods of prevention and treatment is very important, immune infertility technology appears to prevent the Chinese Zokor provides a new way of thinking. Immune infertility is a certain immunologically active substances, directly or indirectly, into a living organism, destroy target organisms own reproductive system, so as to achieve the purpose of reducing its fertility, select the target antigen has been the focus of work, researchers have demonstrated a key role and zona pellucida 3(ZP3) play in the immune infertility. So get Chinese Zokor zona pellucida 3(mZP3) gene and connect it to the eukaryotic expression vector and So get Chinese Zokor zona pellucida 3(mZP3) gene and connect it to the eukaryotic expression vector has become a focus of research. In this study, by constructing a recombinant vector and in vitro protein expression of mZP3, test gene vaccine in mice to lay the foundation for the development of vaccines.In this study, the gene for the purpose of China Zokor mZP3, construct EGFP-mZP3 eukaryotic expression vector, after sequencing correct transfected CHO cells, RT-PCR and WB to detect the expression of target protein. The results indicate that the recombinant plasmid successfully expressed in CHO cells and lay the foundation for the development of vaccines infertility.Construct a new recombinant vector pcDNA3.1-mZP3 to correct the interference of GFP, in front of the target gene sequence a Kozak sequence added to improve the efficiency of its expression, and end of the sequence added 6 His tag for easy detection of the late, because His are small molecules belong to the biochemical nature and will not have an impact on the structure of the target protein, it will not affect their immune titers. After sequencing correct transfected CHO cells in vitro and expression of RT-PCR and WB detection of the target protein. Successfully detected the protein was expressed in CHO and mass production of recombinant plasmids were immunized mice test, mouse sera collected after immunization 14,28,42 d after, and detect the antibody circumstances by Elisa, then carry on the anti-fertility test. The results showed that IgG antibody successfully detected in mice body and also decreased the fertility, which offers the possibility and laid the foundation for the future development of DNA vaccine immune infertility.