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Immunocontraceptive Vaccine for Overpopulation Control in Lagurus lagurus

Author: ZhangAiLian
Tutor: ZhangFuChun
School: Xinjiang University
Course: Zoology
Keywords: Zona pellucida 3 The prokaryotic expression Fusion protein Antiserum DNA vaccines Immune infertility
CLC: S812.6
Type: Master's thesis
Year: 2004
Downloads: 56
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Abstract

Immune infertility vaccine control Lagurus rat fertility studies Lagurus mouse and (Lagurus lagurus) in the case of rodents hamsters Division voles subfamily Lagurus rattus rodents, the distribution area of ??broad reproductive efficiency, high population density, year-round activities bring serious harm to the development and updating of the grassland vegetation is important harmful species in the development of animal husbandry in Xinjiang, seriously restricting the development of animal husbandry in Xinjiang. The rodent control more than with the traditional mechanical hunting, chemical poisoning and other methods, these methods have poor specificity, short off effect, population and rapid recovery shortcomings, and environmental hazards of chemical poisoning methods. With the enhancement of public awareness of environmental protection and animal rights, many countries have started to change the strategy of pest control, immune infertility rodent control as an important research content. Immune infertility technology in controlling wildlife populations showed tantalizing prospect, and has become a current international best potential for population control technology. The zona pellucida in mammals (zona pellucida, ZP) is by oocytes synthesis and secretion, ZP1, ZP2, ZP3 three kinds of sulfated glycoprotein, the sperm must pass through the barriers in the fertilization process. Which ZP3 in the fertilization process has two important roles: First, specific binding of sperm; combined with sperm-induced sperm acrosome reaction. Experimental results demonstrate, mice, pigs and other mammals can block the reproductive process, blocking the combination of sperm with ZP3, ZP3 use of immune infertility technology to control a pest population ideal target antigen. The mouse zona of this study, the main use of the laboratory has been cloned into Lagurus band 3 gene (LZP3) to carry out the grassland lagurus antifertility study. The study mainly include: hamster oocyte of Lagurus pellucida 3 gene prokaryotic expression Preparation of antisera and DNA vaccine in mice and Lagurus mice immunological detection and anti-fertility experiments. First, the use of a prokaryotic system is simple, fast, and convenient features, choose LZP3 fusion protein expressed in the GST system. To improve LZP3 expression LZP3 core fragment removal of the N-terminal signal peptide and C-terminal transmembrane region built into the prokaryotic expression vector pGEX-4T-1 sequence analysis and restriction endonuclease that has been properly constructed pGEX-4T -LZP3. Will build the correct prokaryotic expression plasmid pGEX-4T-LZP3 transformed into E. coli BL21 (DE3), induced by IPTG, SDS-PAGE identification and found that the expression of the fusion protein in 63kDa. Induced by different conditions, the best expression conditions, and then a large number of induced sonicated cells, glutathione Joan lt; WP = 5 gt; agarose bead affinity purification, protein electrophoresis test results show that the per liter of broth can be induced generate a soluble fusion protein was 10 mg. Secondly, the purified of GST-LZP3 protein combined Freund's adjuvant, fractional immune New Zealand white rabbits antiserum final antiserum titer was determined by indirect enzyme-linked immunosorbent assay (ELISA) 1:50 000. E. coli expressed soluble LZP3 protein as antigen using protein immunoblot (Western blot) to detect a polyclonal antibody, the results show that the antiserum having a high specificity. The above study were prepared the high purity LZP3 antigen efficient price, high specificity the rabbit anti LZP3 the antiserum, to immune infertility technical control on grassland lagurus fertility research provides important reagents. The same time, build LZP3 eukaryotic expression plasmid build the LZP3 gene into the eukaryotic expression vector pCDNA3 obtained by restriction endonuclease digestion the pCDNA3-LZP3 recombinant plasmid. Will build a large number of the correct recombinant plasmid extraction and purification and preparation of DNA vaccines. Hamster oocyte Lagurus zona the gene vaccine conducted of grassland lagurus, immunization, to observe the DNA vaccine the grassland lagurus antifertility effect. Female mice after immunization with unimmunized male rats in the same cage, pCDNA3-LZP3 immunohistochemistry: 18 female 13 infertile infertility rate of 73%; empty vector control group: 11 females and 9 birth blank group fertility. Mice after immunization Lagurus cytokines detected T lymphocyte proliferation assays using the MTT assay. The results showed that the mice after immunization Lagurus vivo cytokine levels occurred change, and T-cell proliferation, preliminary DNA vaccine after excitation prairie lagurus specificity of the immune response. The prairie lagurus antifertility experimental results are encouraging, the zona pellucida Lagurus mouse eggs as an immunogen to control the number is feasible. In addition, Lagurus hamster oocyte zona pellucida gene vaccine NIH mice, immunological detection while observing LZP3 ability to produce anti-fertility effect of different kinds of mice. DNA vaccine NIH mice prepared serum immunological detection, anti-fertility in mice after immunization with unimmunized mice in the same cage experiments. Immunological detection by ELISA. Antibody testing results show that the the NIH mice DNA vaccine produced in the body the extremely weak antibody anti LZP3 antifertility experimental results initially showed that LZP3 gene vaccine NIH mice produce anti-fertility effect. This study work for the application Lagurus mouse immune infertility vaccine feasibility study provides a basis for.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > General Animal Science > Grassland Science,prairie school > Grassland protection
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