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Plasmid transfer from Escherichia coli to Bacteroides fragilis: differential expression of antibiotic resistance phenotypes.

机译:从大肠杆菌到脆弱拟杆菌的质粒转移:抗生素抗性表型的差异表达。

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摘要

A unique shuttle plasmid, pDP1, has been constructed to mediate gene transfer between Escherichia coli and the Gram-negative anaerobe Bacteroides fragilis. pDP1 contains the pBR322 replicon and the Bacteroides clindamycin resistance plasmid pCP1 linked to the transfer origin of the broad host range plasmid RK2. pDP1 can be transferred from E. coli to B. fragilis by the RK2 conjugation system even though RK2 itself is not maintained in the Bacteroides recipients. The antibiotic resistance and replication functions of pDP1 have been mapped by deletion analysis, and a 5-kilobase portion of the plasmid has been identified as the essential region for maintenance in Bacteroides. Comparison of the resistance conferred by pDP1 on E. coli and B. fragilis shows that antibiotic resistance genes are expressed differently in aerobic and anaerobic bacteria. These results document the feasibility of gene transfer from E. coli to B. fragilis and demonstrate the usefulness of this conjugation system to study genetic structure and expression in Bacteroides.
机译:已经构建了独特的穿梭质粒pDP1,以介导大肠杆菌和脆弱的革兰氏阴性厌氧细菌之间的基因转移。 pDP1包含pBR322复制子和拟杆菌对克林霉素的抗性质粒pCP1,与宽宿主范围质粒RK2的转移起点相连。 pDP1可以通过RK2缀合系统从大肠杆菌转移到脆弱的B.杆菌中,即使在拟杆菌的受体中不保留RK2本身。 pDP1的抗生素抗性和复制功能已通过缺失分析进行了定位,质粒的5碱基碱基部分已被确定为在拟杆菌中维持的必要区域。 pDP1对大肠杆菌和脆弱型芽孢杆菌的抗性比较表明,好氧和厌氧细菌中的抗生素抗性基因表达不同。这些结果证明了将基因从大肠杆菌转移至脆弱芽孢杆菌的可行性,并证明了该缀合系统对研究拟杆菌中的遗传结构和表达的有用性。

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