Genome AnnouncGenome AnnouncgagaGAGenome Announcements2169-8287American Society for Microbiology1752 N St., N.W., Washington, DC250131334110765genomeA00501-1410.1128/genomeA.00501-14ProkaryotesDraft Whole-Genome Sequences of Nine Non-O157 Shiga Toxin-Producing Escherichia coli StrainsLindseyRebecca L.TreesEijaSammonsScottLoparevVladimirFraceMikeStrockbineNancySabolAshley L.SowersEvanStriplingDevonMartinHaleyKnipeKristenRoweLoriGerner-SmidtPeterCenters for Disease Control and Prevention, Atlanta, Georgia, USAAddress correspondence to Rebecca L. Lindsey, rebecca.lindsey@cdc.hhs.gov.1072014Jul-Aug201424e00501-1412520142362014Copyright © 2014 Lindsey et al.2014Lindsey et al.This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 Unported license.

Shiga toxin-producing Escherichia coli (STEC) is an important food-borne pathogen. Here, we report the draft whole-genome sequences of nine STEC strains isolated from clinical cases in the United States. This is the first report of such information for STEC of serotypes O69, H11, O145:H25, O118:H16, O91:H21, O146:H21, O45:H2, O128:H2, and O121:H19.

cover-dateJuly/August 2014access-typefreeGENOME ANNOUNCEMENT

Shiga toxin-producing Escherichia coli (STEC) is a common cause of food-borne illness. An estimated 265,000 STEC infections occur each year in the United States. Non-O157 STEC strains cause about 64% of these infections, and O157 STEC causes the rest (http://www.cdc.gov/ecoli/general/index.html). The symptoms of STEC infection range from mild, watery to bloody diarrhea, gastroenteritis, hemolytic-uremic syndrome, to death. Most STEC infections are caused by seven serotypes, but >100 STEC serotypes are known to cause illness in humans (1, 2). Only five closed non-O157 STEC genome sequences are publicly available. Four of them (O103, O111, O26, and O145) belong to the most common non-O157 STEC serogroups, and one (O55) is much rarer in prevalence. Here, we report the availability of high-quality draft whole-genome sequences for nine STEC strains that are among the top 15 most common STEC serotypes in prevalence related to human infection in the United States (CDC reference laboratory surveillance, unpublished data). Eight of these draft genome sequences represent STEC serotypes that did not previously have any genome sequences publicly available.

E. coli genomic DNA was extracted according to the manufacturer’s protocol (ArchivePure, 5 Prime, Gaithersburg, MD). DNAs were sheared to 10 kbp or 20 kbp utilizing g-Tubes (Covaris, Inc., Woburn, MA). The 20-kbp sheared products were further size selected utilizing BluePippin size selection (Sage Science, Beverly, MA). The sheared DNAs were used to generate large SMRTbell libraries using the standard library protocols of the Pacific Biosciences DNA template preparation kit (Menlo Park, CA). The finished libraries were bound to proprietary P4 polymerase and sequenced on a PacBio RSII sequencer using C2 chemistry for 120-min movies. The sequence reads were filtered and assembled de novo utilizing the PacBio Hierarchical Genome Assembly Process (3) or a modified Celera Assembler (4). The resulting assemblies were confirmed using OpGen (Gaithersburg, MD) whole-genome maps (WGM). WGM were generated according to the OpGen protocol. The sequences were annotated with the NCBI Prokaryotic Genomes Automatic Annotation Pipeline (5).

A detailed report on further analysis of the draft genome sequences will be included in a future publication.

Nucleotide sequence accession numbers.

The annotated whole-genome E. coli sequences have been deposited in DDBJ/ENA/GenBank under the accession no. JASN00000000 to JASV00000000. The versions described in this paper are the first versions, under the accession no. listed in Table 1.

Accession numbers and assembly metrics of the annotated STEC draft whole-genome sequences

E. coli isolateSerotypeNCBI accession no.No. of contigsGenome size (bp)N50% G+C content
07-3763069:H11JASN00000000195,669,6281,043,19650.7
07-3858O145:H25JASO00000000215,625,860623,35550.7
07-4255O118:H16JASP00000000145,932,5204,019,76750.7
2009C-3740O91:H21JASQ0000000035,026,8614,912,39250.8
2010C-3325O146:H21JASR00000000105,541,5143,834,78150.6
2010C-4211O45:H2JASS00000000215,657,150914,23650.7
2011C-3274O26:H11JAST00000000225,930,1083,776,32250.6
2011C-3317O128:H2JASU00000000165,597,2574,556,44850.7
2011C-3609O121:H19JASV0000000075,412,2723,051,67750.6

Citation Lindsey RL, Trees E, Sammons S, Loparev V, Frace M, Strockbine N, Sabol AL, Sowers E, Stripling D, Martin H, Knipe K, Rowe L, Gerner-Smidt P. 2014. Draft whole-genome sequences of nine non-O157 Shiga toxin-producing Escherichia coli strains. Genome Announc. 2(4):e00501-14. doi:10.1128/genomeA.00501-14.

ACKNOWLEDGMENTS

R.L.L. is a recipient of an ORISE/CDC Postdoctoral Research Fellowship.

The findings and conclusions of this article are those of the authors and do not necessarily represent the views of the Centers for Disease Control and Prevention.

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