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目的研究枯草杆菌对氨茴环霉素（ＡＮＣ）的抗性特征并克隆抗性片段。方法用递增浓度方法在ＬＢ平板上筛选枯草杆菌菌株１８３１对ＡＮＣ的自发抗性突变株，用鸟枪法在枯草杆菌ＢＤ２２４菌株原生质体中克隆抗性重组质粒，提取重组质粒后再次转化敏感菌株以证实克隆片段的抗性功能。结果对ＡＮＣ强抗性突变株ＤＭ１０４对化学特性不相关的化合物如溴化乙锭和诺丹明产生交叉抗性。钙通道阻断剂异搏定使突变株失去对氨茴环霉素的抗性。基因克隆获得了４个带有ＤＭ１０４染色体上不同片段的重组质粒，只有１个片段与已知的枯草杆菌ｂｍｒ基因同源。用重组质粒再次转化敏感菌株证实克隆片段的抗性功能。结论枯草杆菌对ＡＮＣ的抗性呈多重药物抗性特征，这种抗性可能与染色体上多个位点有关
Objective To study the resistance characteristics of Bacillus subtilis to anthracycline (ANC) and clone the resistant fragment. Methods The spontaneous resistance mutants of B. subtilis strain 1831 on ANC were screened on LB plates by using increasing concentration method. The resistant recombinant plasmids were cloned into the protoplasts of Bacillus subtilis BD224 by using the shotgun method. The recombinant plasmids were extracted and transformed into sensitive strains again to confirm Resistant function of cloned fragments. Results Cross-resistance to compounds with strong resistance to ANC, DM104, such as ethidium bromide and normindane, that were not chemically related. Verapamil, a calcium channel blocker, causes mutant strains to lose their resistance to anthracyclines. Gene cloning obtained four with DM104 chromosome different fragments of the recombinant plasmid, only one fragment homologous with the known B. subtilis bmr gene. The recombinant plasmids were transformed into sensitive strains again to confirm the resistance function of the cloned fragments. Conclusion Bacillus subtilis ANC resistance was characterized by multiple drug resistance, which may be related to multiple sites on the chromosome