The expression of Mycobacterium tuberculosis MPT64 protein in transgenic carrots

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.