| Experiment 1 Physical and chemical characteristics of pig ovarian inhibin a subunit were deduced by Antheprot software, including antigenicity profile, hydrophobicity profile, hydrophilic profile, tranmembranous regions profile and solvent accessibility profile. The results indicated that 1-32 and 100-120 regions were two antigenic determinants of inhibin. There was high homogeneity of swine inhibin with human, bovine, ovine inhibin and other protein in the body. So gene encoding inhibin a (1-32) is an candidate one for construct inhibin gene vaccine. Gene encoding porcine inhibin a (1-32) was cloned and recombined with thioredoxin gene on pET-32c plasmid. The recombinant plasmid pEINI-l was expressed in E. coil. DI-IS a~ The expression protein was about 14 KD and had inhibin immunization activity. Unfortunately, the fused protein has high homology with thioredoxin and other proteins in the NR protein bank, so it is not suitable for inhibin antigen. Gene encoding lnhibin a (l挆.32) was recombined with pcDNA3.1 expression vector to construct inhibin gene vaccine pINH. Plasmid pINH mixed with liposome were injected to mouse skeletal muscle at 15~tgI100ii I in group land 25 L?gIlOOL?I in groupll. The first boost was conducted after 3 w of the primary boost. The plasma were prepared before the primary boost, first boost and 3w after the first boost and stored at -20t. Inhibin antibody of the plasma sample was assayed with ELISA. There was inhibin antibody in the plasmas. It indicated that the inhibin gene was expressed in the muscle cells and the expressed material had inhibin antigenecity. The inhibin antibody P/N>2 was looked as positive and the positive ratio was about 30%(6/20). There were little deference of the litter size, birth weight and litter weight between the positive mouse and the control group (p>O.OS). This is the first experiment of using inhibin gene immunization to control animal reproduction and the new approach is a vety promising beginning. In order to improve animal reproductive traits using inhibin gene immunization, these suggestions should be considered for the next experiment, (1) Construct inhibin gene vaccine with higher molecular weight, for example, double inhibin a (1 ?2) gene or recombined inhibin a (1 ?2) 118 gene with the S (small) region of hepatitis B surface antigen,(2)optimizing immunization methods, such as immunization time, dose, injection method and boost. Experiment 2 Chicken bcl-2 gene was recombinant with JLV plasmid and the recombinant expressing plasmid JLVB was induced to chicken granulosa cells with liposome. The granulosa cells were isolated from follicles of laying Romen hens and incubated in 2m1 medium 199 supplementing with 5% calf serum for 3days, and then were transferred by JLVB with 0.0, 0.6, 1.2, 2.4 and 4.81.il of liposome in 24-well plates. The growth and apoptosis of primary cells and subcultured cells were investigated by flow cytometry. The results showed that bcl-2 gene transferred cells grew more quickly (p |
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