CAJ | 학술논문

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 carotaL.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(Mycobacterium tuberculosis(Zopf)Lehmannet Neumann)分泌蛋白MPT64基因的根癌土壤杆菌(Agrobacterium tumefaciens(Smithet 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Southern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。
이용전기인식물생산연제역묘，불단가이개변전통적역묘생산방식화접충수단，이차회대대강저역묘적생산성본。이호라복(Daucus carotaL.var.sativa)무균유묘적자협화하배축위외식체，통과휴대유35S계동자구동적결핵간균(Mycobacterium tuberculosis(Zopf)Lehmannet Neumann)분비단백MPT64기인적근암토양간균(Agrobacterium tumefaciens(Smithet Townsend)Conn)LBA4404적개도진행전화，재사선배양기상유도형성항성유상조직，경배상체발생도경분화득도항성묘，식주이재후생장정황정상。경PCR화Southern등방법감정，학인결핵간균분비단백MPT64기인이정합도호라복적염색체중。Western검측결과표명，재전기인호라복적단백질중함유MPT64분비단백，위진일보연구이용전기인식물연제구복역묘화방치폐결핵적신형역묘제공료신재료。
Oral vaccines produced by transgenic plants would change thetraditional 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.