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为提高外源蛋白的表达量和简化分离纯化过程,使用植物油体表达载体,以花生子叶节为转化受体,通过农杆菌介导将轮状病毒抗原蛋白G3VP7基因开展遗传转化的研究。从转化植株中随机选取11株表现Kan抗性植株进行PCR检测,结果有6株能扩增出特异性条带,阳性率为54.5%。对转基因植株进一步进行PCR-Southern杂交分析,发现转基因植株中有6株PCR反应呈阳性,其中有3株PCR-Southern杂交有特异性目标带出现。结果表明,外源基因已经整合到了花生基因组上。该研究为以植物为载体生产廉价、高效的植物口服疫苗奠定了基础。
In order to improve the expression of exogenous protein and to simplify the process of isolation and purification, the plant oil expression vector was used, the peanut cotyledonary node was used as the transformation receptor, and the genetic transformation of rotavirus antigen G3VP7 gene was carried out by Agrobacterium tumefaciens. A total of 11 Kan-resistant plants were randomly selected from the transformed plants for PCR. As a result, 6 strains were able to amplify specific bands with a positive rate of 54.5%. Further analysis of transgenic plants by PCR-Southern hybridization showed that six of the transgenic plants were positive for PCR, and three of them showed specific target bands by PCR-Southern hybridization. The results showed that foreign genes have been integrated into the genome of peanut. The research laid the foundation for plant-based production of cheap and efficient oral vaccines for plants.