Full-Genome Assembly of Reference Strain Providencia stuartii ATCC 33672.

A member of the normal human gut microflora, Providencia stuartii is of clinical interest due to its role in nosocomial infections of the urinary tract and because it readily acquires antibiotic resistance. Here, we present the complete genome of P. stuartii strain ATCC 33672, consisting of a 4.28-Mbp chromosome and a 48.9-kbp plasmid.

GENOME ANNOUNCEMENT

Providencia stuartii is a frequent cause of urinary tract infections in hospital patients with long-term indwelling catheters (1). In these patients, bacteriuria may rapidly progress to bacteremia (2, 3). Infection with P. stuartii has also been reported to cause peritonitis (4), meningitis (5), pericarditis (6), and infective endocarditis (7). In an unusual case, P. stuartii was identified as the cause of a renal abscess mistakenly attributed to an infection with Pasteurella (8). Additionally, nosocomial dissemination of multidrug-resistant P. stuartii has been reported in burn units (9, 10) and an intensive care unit (10). Members of the genus Providencia harbor resistance to aminopenicillins and narrow-spectrum cephalosporins (11) but also readily acquire resistance to other antibiotics through horizontal transfer (12, 13). This is of great concern to public health due to the ability of P. stuartii to further disseminate antimicrobial resistance genes (13, 14). We present the complete genome of strain ATCC 33672, consisting of a 4.28-Mbp chromosome and a 48.9-kbp plasmid.

High-quality genomic DNA was extracted from a purified isolate using the Qiagen Genome-tip 500. Specifically, a 100-ml bacterial culture was grown to stationary phase and nucleic acid extracted as per the manufacturer’s recommendations. The draft genome of P. stuartii ATCC 33672 included a combination of Illumina (15) and 454 technologies (16). For this genome, we constructed and sequenced a 100-bp Illumina library (372-fold genome coverage) and a long-insert paired-end 454 library (6,689 ± 1,140-bp insert; 5-fold genome coverage). Raw data are available in the Short Read Archive (SRA) under accession numbers SRX687103 (Illumina), SRX687104 (Illumina), and SRX687264 (454).

The 454 paired-end data were assembled in Newbler (16), and those consensus sequences were computationally shredded into 2-kbp overlapping fake reads (shreds). The Illumina sequencing data were assembled with Velvet (17), and the consensus sequences were computationally shredded into 1.5-kbp overlapping shreds. All data were additionally assembled in AllPaths (18), and the consensus sequences were computationally shredded into 5-kbp overlapping shreds. We integrated the 454 Newbler consensus shreds, the Illumina Velvet consensus shreds, AllPaths consensus shreds, and the 454 paired-end library read pairs using parallel Phrap (High Performance Software, LLC). Possible misassemblies were corrected and repeat regions verified using in-house scripts and manual editing in Consed (19–21).

The complete genome assembly of P. stuartii ATCC 33672, including a 4,285,951-bp circular chromosome (41.5% G+C content) and a 48,866-bp circular plasmid (42.8% G+C content), was annotated utilizing an Ergatis-based (22) workflow with minor manual curation. The annotated genome contains 4,094 predicted genes, including 3,926 protein-coding genes, 80 tRNA genes, and 22 rRNA genes. A total of 92 virulence genes were noted, including resistance to fluoroquinolones, β-lactams, and aminoglycosides.

Nucleotide sequence accession numbers.

The nucleotide sequences for P. stuartii ATCC 33672 have been deposited in GenBank under accession numbers CP008920 (chromosome) and CP008919 (plasmid).