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FEMS Microbiol Lett. 2019 Mar 1;366(5). pii: fnz053. doi: 10.1093/femsle/fnz053.

A unique methylation pattern by a type I HsdM methyltransferase prepares for DpnI rare cutting sites in the Pseudomonas aeruginosa PAO1 genome.

Author information

1
Clinical research Group 'Pseudomonas Genomics', OE 6710, Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Lower Saxony, Germany.
2
Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Nobels väg 16, S-171 77 Stockholm, Sweden.

Abstract

A physical genome map of Pseudomonas aeruginosa PAO1 had been constructed in the late 1980s by combinatorial SpeI/DpnI long-range restriction mapping. Whereas SpeI was known to cleave within the infrequent 5'-ACTAGT hexanucleotide, the rare recognition site 5'-Gm6ATC for the methylation-sensitive 4-cutter DpnI in the PAO1 genome remained elusive. Resequencing the PAO1 genome with the methylation-sensitive Single Molecule Real-Time (SMRT) sequencing technology identified DpnI to cleave the PAO1 genome at 15 palindromic GAC(N)6Gm6ATC(N)6GTC recognition sites with the central Gm6ATC palindrome to be methylated on both strands by the clone PAO1-specific type I HsdM superfamily methyltransferase M.PaePAOI (PA2735).

KEYWORDS:

bacteriology; genome reconstruction; methylation; restriction modification

PMID:
30865771
DOI:
10.1093/femsle/fnz053

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