Identification of fbpA gene transformed to tomato plants encoding secreted Ag85A of BCG

Objective To identify the fbpA gene transformed to tomato plants encoding secreted Ag85A from BCG.Methods Rapid PCR amplification of boiled leaves and PCR detection of total cellular DNA from tomato plants,and detection of transformed tomato plants by Southern blot hybridization were used to identify the integration of fbpA in the tomato genomic DNA.Western blot hybridization was used to identify the expression of secreted Ag85A in the tomato fruits.Results 21 positive shoots were identified by rapid PCR amplification of boiled leaves.9 plants of them carried the fbpA gene by PCR detection of total cellular DNA.It was conformed that the fbpA gene was integrated in the genomic DNA of these 9 plants.The expression of secreted Ag85A was identified in fruits of 5 tomato plants by Western blot hybridization,and the specific immunization were observed with the serum of patients from tuberculosis.Conclusion The fbpA gene is integrated in tomato genomic DNA and the secreted Ag85A was expressed in the tomato fruits.