NR-42890??Salmonella enterica subsp. enterica, 14028s (Serovar Typhimurium) Single-Gene Deletion Mutant Library, Plate SGD_156/157_Cm(Mutant Bacteria)|Salmonella enterica subsp. enterica| 14028s (Serovar Typhimurium) Single-Gene Deletion Mutant Library, Plate SGD_156/157_Cm|-80°C or colder|M McClellandAcknowledgment for publications should read “The following reagent was obtained through BEI Resources, NIAID, NIH: Salmonella enterica subsp. enterica, Strain 14028s (Serovar Typhimurium) Single-Gene Deletion Mutant Library, Plate SGD_156/157_Cm, NR-42890.”|
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.
The Salmonella
enterica (S. enterica) subsp. enterica, strain 14028s (serovar Typhimurium) targeted
single-gene deletion (SGD) mutant library contains a total of 3,773 individual
genes deleted simultaneously across two collections of mutants differentiated
by kanamycin or chloramphenicol resistance. The
chloramphenicol-resistant mutant collection contains 3,376 mutants distributed
among eleven 96-well plates, in which a single gene is replaced by a cassette
conferring the chloramphenicol resistance gene, and includes 4 double mutants
that contain both kanamycin and chloramphenicol cassettes. Deletions were confirmed by the depositor.
Genes were targeted for deletion by primers
designed to preserve the first and last 30 bases of each deleted gene. Gene replacement followed a modified Lambda-Red technique, with an added T7 RNA
polymerase promoter positioned in plasmid pCLF3 to
generate a gene-specific transcript from the Salmonella genome directly downstream of each mutant.
Note: The strain designation on the plate, strain CDC 6516-60,
is incorrect. The correct strain designation is strain 14028s. S. enterica
subsp. enterica, strain 14028s was originally known as strain
14028. A variant of the original strain with a rough colony morphology
was designated 14028r and the original smooth strain was renamed 14028s.
Strain 14028 is a descendent of strain CDC 6516-60 which was isolated from
pools of hearts and livers of 4-week-old chickens. The complete
genome of S. enterica subsp. enterica, strain 14028s
(GenBank: CP001363.1)
and plasmid (GenBank: CP001362.1)
sequences are available.
Each inoculated well of the 96-well plate
contains approximately 50 ?L of culture in Luria Bertani (LB) broth containing 20
?g/mL chloramphenicol supplemented with 10% glycerol.
Production
in the 96-well format has increased risk of cross-contamination between
adjacent wells. Individual clones should be purified (e.g. single colony
isolation and purification using good microbiological practices) and
sequence-verified prior to use. BEI Resources does not confirm or validate
individual mutants provided by the contributor.