摘要
目的了解卡介苗分泌型Ag85A基因fbpA在番茄中转化的情况。方法采用叶片煮沸快速PCR法检测fbpA基因;以番茄总DNA为模板,PCR法扩增目的基因fbpA;采用Southern blot检测目的基因fbpA在番茄染色体DNA中的整合情况,Western blot检测分泌型Ag85A在番茄果实中的表达情况。结果叶片煮沸快速PCR检出21株fbpA阳性转化不定芽,以番茄总DNA为模板的PCR法扩增出其中9株转化植株带有目的基因fbpA。Southern blot证实这9株番茄染色体DNA中整合了外源目的基因fbpA,Western blot检测到其中5株番茄果实中表达的卡介苗分泌型Ag85A与结核病人血清发生特异性免疫反应。结论番茄染色体DNA中整合了外源目的基因fbpA,果实中有该基因产物分泌型Ag85A表达。
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.
出处
《中国病原生物学杂志》
CSCD
2007年第4期254-256,共3页
Journal of Pathogen Biology
