摘要
利用转基因植物生产研制疫苗 ,不但可以改变传统的疫苗生产方式和接种手段 ,而且会大大降低疫苗的生产成本。以胡萝卜 (DaucuscarotaL .var.sativa)无菌幼苗的子叶和下胚轴为外植体 ,通过携带有 35S启动子驱动的结核杆菌 (Mycobacteriumtuberculosis (Zopf)LehmannetNeumann)分泌蛋白MPT6 4基因的根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)LBA44 0 4的介导进行转化 ,在筛选培养基上诱导形成抗性愈伤组织 ,经胚状体发生途径分化得到抗性苗 ,植株移栽后生长情况正常。经PCR和Southern等方法鉴定 ,确认结核杆菌分泌蛋白MPT6 4基因已整合到胡萝卜的染色体中。Western检测结果表明 ,在转基因胡萝卜的蛋白质中含有MPT6 4分泌蛋白 ,为进一步研究利用转基因植物研制口服疫苗和防治肺结核的新型疫苗提供了新材料。
Oral vaccines produced by transgenic plants would change the traditional means of production and inoculation of vaccines and the cost of vaccine production would be reduced greatly. In the experiments, hypocotyls and cotyledons of carrot ( Daucus carota L. var. sativa) were infected with Agrobacterium tumefaciens (Smith et Townsend) Conn LBA4404 containing Mycobacterium. tuberculosis (Zopf) Lehmann et Neumann MPT64 gene under the control of the 35S promoter of cauliflower mosaic virus. After two days coculture, the explants were transferred to MS selection media which contained different concentrations of kanamycin and carbenicillin. The regenerated plants with kanamycin resistance were obtained through somatic embryogenesis from the embryogenic calli formed on the selection media. Some of the plants have been transplanted and grew well in phytotron. PCR and Southern blot analyses of carrot DNA confirmed that the MPT64 gene has been introduced into the plant genome. The results of Western blot showed that the MPT64 protein have been expressed in some transgenic plants. Therefore, the transgenic plants should provide a valuable tool for the development of edible oral vaccines.
基金
上海市自然科学基金! (99ZF14 0 48)
植物分子遗传国家重点实验室资助&&
