Abstract:

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.

利用转基因胡萝卜表达肺结核疫苗
王凌健 倪迪安 陈永宁 李忠明*

（中国科学院上海植物生理研究所，上海200032）

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(Daucus carota L.var.sativa)无菌幼苗的子叶和下胚轴为外植体，通过携带有35S启动的结核杆菌（Mycobacterium tuberculosis（Zopf）Lehmann et Neumann）分泌蛋白MPT64基因的根癌壤杆菌（Agrobacterium tumefaciens(Smith et Townsend)Conn）LBA4404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Southerm 等方法鉴定，确认结核杆菌分泌蛋白MPT64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MPT64 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。
关键词： 转基因胡萝卜；口服疫苗；结核杆菌MPT64  分泌蛋白

Key words: transgenic carrot, oral vaccine, Mycobacterium tuberculosis MPT64 protein