title: [Expression of Streptococcus mutans surface protein PAcP and cholera toxin B subunit fusion gene in transgenic tomato].
abstract: OBJECTIVE
  To detect the protein expression level of foreign fused gene in the first generation of genetically modified tomatoes transformed with fused gene of antigen epitope PacP in surface protein of Streptococcus mutans and cholera toxin B subunit.
  
  
  METHODS
  The total DNA in tomatoes was extracted and PCR was applied to screen the first generation of transgenic tomatoes carrying Streptococcus mutans surface protein coding PAcP with cholera toxin B subunit fusion gene. Total proteins were extracted and quantitatively tested with BAC kit. Foreign fused protein expression level was analysed by Western blotting and ELISA.
  
  
  RESULTS
  Ten DNA samples with the full length of 1.6 kb foreign fused DNA fragment was detected by PCR among all 18 samples. The crude protein in the transgenic tomato fruit tissue was 3.93 mg/mL. Compared with the normal tomatoes, Western blotting showed that the transgenic tomato protein contained a distinct protein band with a molecular weight about 60 kD. The results of ELISA suggested that fused foreign protein level was up to 0.18% of the total soluble protein in genetically modified tomato fruit.
  
  
  CONCLUSIONS
  The target protein encoded by fused foreign gene of antigen epitope PacP in surface protein and cholera toxin B subunit was expressed in genetically modified tomatoes fruit.
authors.affiliation: School of Stomatology, Zunyi Medical University. Zunyi 563000;Guizhou Province, China.E-mail:gylookfish@126.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Gu
authors.forename: Jian-guo
authors.initials: JG
authors.lastname: Liu
authors.forename: Wei-wei
authors.initials: WW
authors.lastname: Guan
authors.forename: Zhu
authors.initials: Z
authors.lastname: Chen
authors.forename: Guo-hui
authors.initials: GH
authors.lastname: Bai
authors.forename: Lin
authors.initials: L
authors.lastname: Tang
authors.forename: Xiao-yan
authors.initials: XY
authors.lastname: Guan
authors.forename: Yuan
authors.initials: Y
authors.lastname: Tian
authors.forename: Qi
authors.initials: Q
authors.lastname: Han
mesh_terms: D015153:Blotting, Western; D002772:Cholera Toxin; D004797:Enzyme-Linked Immunosorbent Assay; D018551:Solanum lycopersicum; D008565:Membrane Proteins; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D013295:Streptococcus mutans
chemical_list: D008565:Membrane Proteins; D002772:Cholera Toxin