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Journal of Bacteriology, September 2009, p. 5717-5723, Vol. 191, No. 18
0021-9193/09/$08.00+0     doi:10.1128/JB.00325-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Optical Mapping Reveals a Large Genetic Inversion between Two Methicillin-Resistant Staphylococcus aureus Strains{triangledown} ,{dagger}

Sanjay K. Shukla,1* Jennifer Kislow,1 Adam Briska,2 John Henkhaus,2 and Colin Dykes2

Marshfield Clinic Research Foundation, 1000 North Oak Avenue, Marshfield, Wisconsin 54449,1 OpGen Inc., 708 Quince Orchard Road, Gaithersburg, Maryland 208782

Received 9 March 2009/ Accepted 16 June 2009

Staphylococcus aureus is a highly versatile and evolving bacterium of great clinical importance. S. aureus can evolve by acquiring single nucleotide polymorphisms and mobile genetic elements and by recombination events. Identification and location of novel genomic elements in a bacterial genome are not straightforward, unless the whole genome is sequenced. Optical mapping is a new tool that creates a high-resolution, in situ ordered restriction map of a bacterial genome. These maps can be used to determine genomic organization and perform comparative genomics to identify genomic rearrangements, such as insertions, deletions, duplications, and inversions, compared to an in silico (virtual) restriction map of a known genome sequence. Using this technology, we report here the identification, approximate location, and characterization of a genetic inversion of ~500 kb of a DNA element between the NRS387 (USA800) and FPR3757 (USA300) strains. The presence of the inversion and location of its junction sites were confirmed by site-specific PCR and sequencing. At both the left and right junction sites in NRS387, an IS1181 element and a 73-bp sequence were identified as inverted repeats, which could explain the possible mechanism of the inversion event.

* Corresponding author. Mailing address: Marshfield Clinic Research Foundation, 1000 North Oak Avenue-MLR, Marshfield, WI 54449. Phone: (715) 389-5363. Fax: (715) 389-3808. E-mail: shukla.sanjay{at}mcrf.mfldclin.edu

{triangledown} Published ahead of print on 19 June 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, September 2009, p. 5717-5723, Vol. 191, No. 18
0021-9193/09/$08.00+0     doi:10.1128/JB.00325-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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