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目的:以卡介苗(BCG)和结核分枝杆菌(MTB)国际标准无毒株H37Ra(H37Ra)的原生质体为亲本制备融合菌株。方法:制备及鉴定BCG和H37Ra的原生质体;对荧光染色标记的亲本原生质体进行电融合,制备融合菌株,同时优化其制备和再生培养条件。结果:成功制备BCG和H37Ra的原生质体;FDA标记BCG原生质体呈黄绿色荧光,罗丹明B标记H37Ra原生质
OBJECTIVE: To prepare fusion strains using protoplasts of Bacillus Calmette-Guerin (BCG) and Mycobacterium tuberculosis (MTB) as an international standard non-virulent strain H37Ra (H37Ra) as their parents. Methods: Protoplasts of BCG and H37Ra were prepared and identified. Fluorescently stained parental protoplasts were electrofused to prepare fusion strains, and their preparation and regeneration conditions were optimized. Results: The protoplasts of BCG and H37Ra were successfully prepared. The protoplasts of FDA labeled BCG were yellow-green fluorescence and rhodamine B labeled H37Ra protoplasts