A Novel Plasmid pALWVS1.4 from Acinetobacter lwoffii Strain VS15, Carrying the Chloramphenicol Resistance Gene
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A Novel Plasmid pALWVS1.4 from Acinetobacter lwoffii Strain VS15, Carrying the Chloramphenicol Resistance Gene A. Y. Ermakovaa, A. V. Beletskya, A. V. Mardanova, M. A. Petrovab, N. V. Ravina, and A. L. Rakitina, * a
Institute of Bioengineering, Research Center of Biotechnology, Russian Academy of Sciences, Moscow, 119071 Russia bInstitute of Molecular Genetics, Russian Academy of Sciences, Moscow, 123182 Russia *e-mail: [email protected] Received April 19, 2020; revised May 7, 2020; accepted May 8, 2020
Abstract—Sequencing and analysis of the genome of the chloramphenicol-resistant strain Acinetobacter lwoffii VS15, isolated from permafrost, revealed a circular plasmid 11964 bp in length, designated pALWVS1.4. Apart from the genes supporting the plasmid maintenance and mobilization, pALWVS1.4 contains the cflA gene encoding the membrane protein of the MFS transporter family. Close homologues of cflA were found in bacteria of the genus Psychrobacter, but were absent in the Acinetobacter genomes. The cflA gene is flanked by two copies of IS elements of the IS4 family, indicating that it was acquired by A. lwoffii VS15 via horizontal transfer within this putative composite transposon. A recombinant Escherichia coli strain expressing CflA was obtained and it was shown that under conditions of induced expression, the strain was resistant to chloramphenicol. Keywords: Acinetobacter, permafrost, antibiotic resistance, plasmid, horizontal gene transfer DOI: 10.1134/S0026261720050070
Acinetobacter bacteria are ubiquitous due to their ability to adapt to diverse ecological niches (Touchon et al., 2014). Some species, primarily A. baumannii, are clinically important pathogens; others, for example, A. calcoaceticus, A. johnsonii, A. haemolyticus, and A. lwoffii, are widespread both in nature and in the clinical setting, while most species live in aquatic ecosystems and soils (Doughari et al., 2011). Such plasticity of Acinetobacter strains is largely provided by a wide variety of plasmids. The spread of the antibiotic resistance determinants that are part of various mobile elements and the associated increase in the number of resistant clinical Acinetobacter strains is an important problem for medicine (Da Silva and Domingues, 2016). Both environmental Acinetobacter strains and other types of bacteria can be sources of mobile elements with resistance genes. Currently, four main mechanisms of bacterial resistance to antibiotics have been identified: (1) modification of the target of antibiotic action, (2) enzymatic inactivation of the antibiotic, (3) decrease in the permeability of microbial cell membranes, and (4) active removal of antimicrobial agents from the cell using various transporters (Nikaido, 2009; van Hoek et al., 2011). The resistance mechanism due to active removal of antibiotic molecules from the cell is based on the work of a specialized set of proteins that form transmembrane transporters. Such transmembrane pumps are
capable of transporting toxic substances and xenobiotics, including ant
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