Cloning of Helicobacter pylori urease subunit B gene and its expression in tobacco (Nicotiana tabacum L.).
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.
Gu
Qing
Q
State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310012, China.
Han
Ning
N
Liu
Jianyi
J
Zhu
Muyuan
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D018744:DNA, Plant; D005822:Genetic Vectors; D016480:Helicobacter pylori / Q000201:enzymology*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics; D014026:Nicotiana / Q000235:genetics*; D014510:Urease / Q000235:genetics*
D018744:DNA, Plant; D011994:Recombinant Proteins; D014510:Urease