轮状病毒抗原蛋白G3VP7基因在花生中遗传转化的研究  

Study on Genetic Transformation of Antigen G3VP7 Gene of Human Rotavirus in Peanut

作　　者：贾宇臣[1] 赵凯[2] 薛昕[3] 李薇[3] 李少伟[1] 刘德虎[4] 

机构地区：[1]内蒙古医学院分子生物学研究中心,呼和浩特010059 [2]内蒙古呼和浩特市第一医院儿内科,呼和浩特010030 [3]内蒙古医学院生物化学与分子生物学教研室,呼和浩特010110 [4]中国农业科学院生物技术所,北京100081

出　　处：《生物医学工程学杂志》2012年第2期328-331,共4页Journal of Biomedical Engineering

摘　　要：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 increase the expression level of target gene and to simplify the purifying process of separation and purification,we performed the transgenetic research of antigen VP7 gene into peanut via Agrobacterium tumefaciens.The plant binary expression vector is pBOG3VP7 harboring fusion gene oleosin-vp7,which is promoted by ole-promoter.Cotyledon nodes were used as transformation recipients.Transformed individuals were obtained through selection on medium containing 125 mg L-1 Kan.Integration of transgenes was assessed by PCR amplification and PCR-Southern blot hybridization.Taking pBOG3VP7 plasmid as positive control,non-transformed peanut as negative control.6 plants among 11 plants grown up through seletion medium were detected by PCR and the rate of positive plants is 54.5%.PCR positive plants were further analysed by PCR-Southern blot hybridization.The results showed that 3 plants have DNA bloting bands.The results also showed that the foreign gene was integrated into genome of transformed peanuts.Elevated expression of rotavirus VP7 antigen in transgenic peanuts was a critical factor in the development of efficient and cheap plant oral vaccine.

关 键 词：花生 VP7基因 人源轮状病毒 植物口服疫苗 遗传转化 

分 类 号：Q943.2[生物学—植物学] R392.1[医药卫生—免疫学]