Shanghai Journal of Stomatology ›› 2014, Vol. 23 ›› Issue (3): 261-265.

• Basic Study • Previous Articles     Next Articles

Expression of Streptococcus mutans surface protein PAcP and cholera toxin B subunit fusion gene in transgenic tomato

GU Yu1,2, LIU Jian-guo1,2,GUAN Wei-wei1, CHEN Zhu1,3, BAI Guo-hui1,2, TANG Lin1,3, GUAN Xiao-yan1,2, TIAN Yuan1,2, HAN Qi1   

  1. 1.School of Stomatology, Zunyi Medical University. Zunyi 563000;
    2. Special Key Laboratory of Oral Diseases Research,Higher Education Institution In Guizhou Province. Zunyi 563000;
    3. Stomatological Hospital of Guiyang. Guiyang 550002, Guizhou Province, China
  • Received:2013-10-11 Online:2014-06-20 Published:2014-09-09
  • Supported by:
    Supported by National Natural Science Foundation of China (30160086), Science and Technology Support Project of Guizhou Province [SY(2012)3086], The Sixth Batch of Technology Innovation Team Project in Guizhou Province[(2013)4026] and Science and Technology Foundation of Guizhou Province [LKZ(2011)41].

Abstract: PURPOSE: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.

Key words: Streptococcus mutans, Cholera toxin B Subunit, Transgenic tomatoes, Edible vaccine

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