Gu Qing, Han Ning, Liu Jianyi, Zhu Muyuan
State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310012, China.
Plant Cell Rep. 2005 Nov;24(9):532-9. doi: 10.1007/s00299-005-0962-8. Epub 2005 Aug 26.
Vaccines produced by transgenic plants would have the potential to change the traditional means of production and inoculation of vaccines, and to reduce the cost of vaccine production. In the present study, an UreB antigen gene from a new Helicobacter pylori strain ZJC02 was cloned into the binary vector pBI121 which contains a CaMV35S promoter and a kanamycin resistance gene, and then transformed UreB into tobacco leaf-disc by Agrobacterium-mediated method. A total of 50 regenerated plants with kanamycin resistance were obtained in the selection media. The 35 putative transgenic individuals were tested and verified the presence and integration of the UreB into the nuclear genome of tobacco plants by PCR, PCR-southern, and Southern analyses. Expression of UreB gene in the tobacco plants was confirmed by RT-PCR and Western Blot analysis using polyclonal human antiserum. To our knowledge, this is the first report of the expression of Helicobacter pylori UreB antigen gene in a plant system, suggesting a major step in the production of plant-based vaccines for Helicobacter pylori.
转基因植物生产的疫苗有可能改变传统的疫苗生产和接种方式，并降低疫苗生产成本。在本研究中，将一株新的幽门螺杆菌ZJC02的脲酶B抗原基因克隆到含有CaMV35S启动子和卡那霉素抗性基因的双元载体pBI121中，然后通过农杆菌介导的方法将脲酶B转化到烟草叶盘。在选择培养基上共获得了50株具有卡那霉素抗性的再生植株。通过PCR、PCR- Southern和Southern分析对35个推定的转基因个体进行检测，证实脲酶B已存在并整合到烟草植株的核基因组中。使用多克隆人抗血清通过RT-PCR和Western Blot分析证实了脲酶B基因在烟草植株中的表达。据我们所知，这是幽门螺杆菌脲酶B抗原基因在植物系统中表达的首次报道，表明在生产幽门螺杆菌植物疫苗方面迈出了重要一步。
