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NR-50352   Escherichia coli, Escherichia coli - Staphylococcal Shuttle Vectors and Hosts
(Assays/Panels)
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Description: Escherichia coli - Staphylococcal Shuttle Vectors and Hosts
Organism: Escherichia coli
Biosafety Level: 1
Availability Status: In Stock
Store at: -60°C or colder
Contributor: JL Bose, EV Stabb
Comments:
Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment.

NR-50352 is a kit containing two vectors that remain stable during both in vitro and in vivo experiments without the requirement of antibiotics, and two Escherichia coli (E. coli) hosts for genetic manipulations prior to transfer into Staphylococcus aureus (S. aureus). The shuttle vectors are based on the LAC-p01 plasmid. pKK22 is designed to be used with USA300 strains that contain LAC-p01 and will render the strains isogenic. pKK30 is a variant of pKK22 in which open reading frames (ORFs) not needed for vector maintenance have been deleted and is intended for use in Staphylococcal cells not containing LAC-p01. Both vectors contain a single trimethoprim resistance cassette that is functional in both E. coli and Staphylococcus species. Additionally, they contain the E. coli R6Kγ origin of replication, which requires pir+ cells for replication thus, DH5αλpir and the pir+ dam- dcm- strain, GM2163λpir, are provided as host strains.  The complete nucleotide sequences of pKK22 and pKK30 are available (GenBank: KX085042 and KX085043, respectively)

Catalog Number
Vector or Host
Comments
NR-50348 pKK22 For use in E. coli DH5αλpir or GM2163λpir or S. sureus USA300 strains containing LAC-p01
NR-50349 pKK30 pKK30 is a variant of pKK22, for use in E. coli DH5αλpir or GM2163λpir or Staphylococcus species not containing LAC-p01
NR-50350 E. coli DH5αλpir Host strain containing the pir genes for performing genetic manipulations prior to transfer into Staphylococcus (F-ϕdlacZ ΔM15 ΔacZYA-argF U169 deoR supE44 hsdR17 recA1 endA1 gyrA96 thi-1 relA1)
NR-50351 E. coli GM2163λpir Host strain containing the pir genes for performing genetic manipulations. This strain is also a dam and dcm methylase mutant for transfer of plasmids into Staphylococcus isolates that do not accept E. coli DNA easily (F-ara-14 leuB6 fhuA31 lacY1 tsx78 glnV44 galK2 galT22 mcrA dcm-6 hisG4 rfbD1 rpsL136 dam13::Tn9 xylA5 mtl-1 thi-1 mcrB1 hsdR2 λpir)

This strain was developed by Krute, et al., at the Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, PubMed:  27637878.

  NR-50352 is comprised of 4 vials containing:
NR-50348: 0.5 mL of vector pKK22, recombinant in E. coli DH5αλpir, in Tryptic Soy broth containing 10 µg/mL trimethoprim supplemented with 10% glycerol.
NR-50349: 0.5 mL of vector pKK30, recombinant in E. coli DH5αλpir, in Tryptic Soy broth containing 10 µg/mL trimethoprim supplemented with 10% glycerol.
NR-50350: 0.5 mL of E. coli DH5αλpir in Tryptic Soy broth supplemented with 10% glycerol.
NR-50351: 0.5 mL of E. coli GM2163λpir in Tryptic Soy broth supplemented with 10% glycerol.
Citations: Acknowledgment for publications should read “The following reagent was contributed by Dr. J. L. Bose for distribution by BEI Resources, NIAID, NIH: Escherichia coli – Staphylococcal Shuttle Vectors and Hosts, NR-50352.”
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  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.

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