Our lab is active in generating new genetic tools for studying Staphylococcus and related bacteria. Below is a short list of some of our key resources.
The following resources have been made freely-available through beiresources.org:
NTML: The Nebraska Transposon Mutant Library is a collection of 1,952 defined bursa aurealis transposon insertions in the chromosome of the CA-MRSA derivative JE2. For more information please see: PMID: 23404398 and http://app1.unmc.edu/fgx/tools.html
NTML Toolkit: To increase the utility of the library, Dr. Bose created a toolkit that allows for the replacement of the NTML transposon with an unmarked deletion, one of three alternate antibiotic selectable markers or five different codon-optimized fluorescent proteins. For more information please see: PMID 23354696 and http://app1.unmc.edu/fgx/tools.html
pJB38: This plasmid is for allelic exchange in Staphylococcus species. It is a hybrid of the pCL10 temperature-sensitive plasmid with the convenience of an anhydrotetracycline-inducible counter-selection taken from pKOR1. We know that it has been successfully used by many labs working with both S. aureus and S. epidermidis. For more information please see: PMID 23354696 and http://app1.unmc.edu/fgx/tools.html
pKK22 and pKK30 stable plasmids:Plasmid maintenance by bacteria can be unpredictable without selective pressure. This is especially a concern during in vivo studies. To circumvent this we created plasmids that remain stable in Staphylococcus species without antibiotic selection. We have tested them in vitro with both S. aureus and S. epidermidis. Furthermore, we have shown them to be stable for at least 7 days in vivo. A colleague has used pKK22 for 14 days. Both plasmids and their E. coli host strains (Thanks to my old mentor Eric Stabb at UGA for sharing) are soon to be available through BEI.
The following resources are available directly through us:
Red Staph: KUB7 is a derivative of the CA-MRSA strain LAC that has a constitutively-expressed codon-optimized DsRed integrated into the chromosome. This strain is simply referred to as “Red Staph” by our colleague and has worked well for imaging and tracking S. aureus during infection. For more information, see PMID 27032615
Codon-optimized lacZ: Along with Dr. Kelly Rice (Univ of Florida), we have constructed a S. aureus codon-optimized lacZ gene with custom multiple cloning site and removable optimized RBS. This construct is much improved over previous lacZ genes used in S. aureus and dramatically enhances reporter output. We currently have it in pJB185. For more information, see PMID 28031278