pHNAH67 lncHI2 plasmid廣宿主質粒載體

Bacteriophages are the most abundant biological entities on earth and may play an important role in the transmission of antibiotic resistance genes (ARG) from host bacteria. Although the specialized transduction mediated by the temperate phage targeting a specific insertion site is widely explored, the carrying characteristics of “transducing particles” for different ARG subtypes in the process of generalized transduction remains largely unclear. Here, we isolated a new T4-like lytic phage targeting transconjugant Escherichia coli C600 that contained plasmid pHNAH67 (KX246266) and encoded 11 different ARG subtypes. We found that phage AH67C600_Q9 can misload plasmid-borne ARGs and package host DNA randomly. Moreover, for any specific ARG subtype, the carrying frequency was negatively correlated with the multiplicity of infection (MOI). Further, whole genome sequencing (WGS) identified that only 0.338% (4/1183) of the contigs of an entire purified phage population contained ARG sequences; these were floR, sul2, aph(4)-Ia, and fosA. The low coverage indicated that long-read sequencing methods are needed to explore the mechanism of ARG transmission during generalized transduction.

Map:
Gene map of lncHI2 plasmid pHNAH67. (A) Full annotated plasmid map. (B) Partial map bp 82,900 to 105,000 indicating the positions of ARG subtypes floR, sul2, aph(4)-Ia, acc(3)-IV, fosA3, blaCTX-M-65, aac(6′)-lb-cr, blaOXA-1, catB3, arr3, and sul1.

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