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Published on: August 20, 2021
Complete nucleotide sequence of Klebsiella pneumoniae multiresistance plasmid pJHCMW1
Renee Sarno1, Glen McGillivary, David J Sherratt
1Department of Biological Science, Institute of Molecular Biology and Nutrition, College of Natural Science and Mathematics, California State University Fullerton, Fullerton, California 92834-6850, USA.
Abstract:
The multiresistance plasmid pJHCMW1, harbored by a clinical Klebsiella pneumoniae strain isolated from a neonate with meningitis, was sequenced. A circular sequence of 11,354 bp was generated, of which 7,993 bp make up Tn1331, a transposon including the antibiotic resistance genes aac(6')-Ib, aadA1, bla(OXA-9), and bla(TEM-1). The gene aac(6')-Ib is included in a gene cassette, and both aadA1 and bla(OXA-9) are included in a single-gene cassette that may have arisen as a consequence of a recombination event involving two integrons. The pJHCMW1 plasmid replicates through a ColE1-like RNA-regulated mechanism, includes a functional oriT, and two loci with similarity to XerCD site-specific recombination target sites involved in plasmid stabilization by the resolution of multimers. One of these two loci, mwr, is active and has been the subject of previous studies, and the other, dxs, is not functional but binds the recombinase XerD with low affinity. Two additional open reading frames were identified, one with low similarity to two hypothetical membrane proteins from Mycobacterium tuberculosis and Mycobacterium leprae and the other with low similarity to psiB, a gene encoding a function that facilitates the establishment of the transferring plasmid in the recipient bacterial cell during the process of conjugation.
Insights
The multiresistance plasmid pJHCMW1 from a meningitis-causing Klebsiella pneumoniae strain was sequenced. It contains the Tn1331 transposon with multiple antibiotic resistance genes and replication/stabilization mechanisms.
Area of Science:
- Microbiology
- Genomics
- Molecular Biology
Background:
- Clinical isolates of Klebsiella pneumoniae can harbor mobile genetic elements conferring antibiotic resistance.
- Neonatal meningitis is a serious infection requiring effective antimicrobial treatment.
Purpose of the Study:
- To sequence and characterize the multiresistance plasmid pJHCMW1 from a clinical Klebsiella pneumoniae isolate.
- To identify antibiotic resistance genes and genetic elements involved in plasmid replication and stability.
Main Methods:
- Whole-genome sequencing of the pJHCMW1 plasmid.
- Bioinformatic analysis to identify genes, transposons, and regulatory elements.
- Comparison with known genetic elements and databases.
Main Results:
- The 11,354 bp plasmid pJHCMW1 contains the Tn1331 transposon (7,993 bp) with four antibiotic resistance genes: aac(6')-Ib, aadA1, bla(OXA-9), and bla(TEM-1).
- The plasmid utilizes a ColE1-like replication mechanism and possesses a functional oriT for conjugation.
- Two XerCD site-specific recombination target sites (mwr and dxs) were identified for plasmid stabilization, with mwr being active and dxs non-functional.
- Two additional open reading frames with similarities to hypothetical membrane proteins and a conjugation-related gene (psiB) were found.
Conclusions:
- The pJHCMW1 plasmid is a significant vehicle for the dissemination of multiple antibiotic resistance genes in Klebsiella pneumoniae.
- Understanding the plasmid's genetic architecture provides insights into its replication, stability, and potential for horizontal gene transfer.
- This characterization aids in comprehending the molecular basis of antibiotic resistance in clinical pathogens.
