Product Sheet Info

Product Information Sheet for NR-42892

Salmonella enterica subsp. enterica, Strain Material Provided: 14028s (Serovar Typhimurium) Single- Each inoculated well of the 96-well plate contains approximately 50 µL of culture in Luria Bertani (LB) broth Gene Deletion Mutant Library, Plate containing 20 µg/mL chloramphenicol supplemented with 10% SGD_160/161_Cm glycerol.

Catalog No. NR-42892 Packaging/Storage: NR-42892 was packaged aseptically in a 96-well plate. The product is provided frozen and should be stored at -80°C or For research use only. Not for human use. colder immediately upon arrival. For long-term storage, the vapor phase of a liquid nitrogen freezer is recommended. Contributor: Freeze-thaw cycles should be avoided. Michael McClelland, Professor, Scientific Director, Vaccine Research Institute of San Diego, San Diego, California, USA Growth Conditions: Media: Manufacturer: LB broth or agar containing 20 µg/mL chloramphenicol BEI Resources Incubation: Temperature: 37°C Product Description: Atmosphere: Aerobic Production in the 96-well format has increased risk of cross- Propagation: contamination between adjacent wells. Individual clones 1. Scrape top of frozen well with a pipette tip and streak onto should be purified (e.g. single colony isolation and purification agar plate. using good microbiological practices) and sequence-verified 2. Incubate the plates at 37°C for 24 hours. prior to use. BEI Resources does not confirm or validate individual mutants provided by the contributor. Citation: Acknowledgment for publications should read “The following The Salmonella enterica (S. enterica) subsp. enterica, strain reagent was obtained through BEI Resources, NIAID, NIH: 14028s (serovar Typhimurium) targeted single-gene deletion Salmonella enterica subsp. enterica, Strain 14028s (Serovar (SGD) mutant library contains a total of 3,773 individual genes Typhimurium) Single-Gene Deletion Mutant Library, Plate deleted simultaneously across two collections of mutants SGD_160/161_Cm, NR-42892.” differentiated by kanamycin or chloramphenicol resistance.1,2 The chloramphenicol-resistant mutant collection contains Biosafety Level: 2 3,376 mutants distributed among eleven 96-well plates, in Appropriate safety procedures should always be used with this which a single gene is replaced by a cassette conferring the material. Laboratory safety is discussed in the following chloramphenicol resistance gene, and includes 4 double publication: U.S. Department of Health and Human Services, mutants that contain both kanamycin and chloramphenicol Public Health Service, Centers for Disease Control and cassettes. Deletions were confirmed by the depositor.1,2 Prevention, and National Institutes of Health. Biosafety in Microbiological and Biomedical Laboratories. 5th ed. Genes were targeted for deletion by primers designed to Washington, DC: U.S. Government Printing Office, 2009; see preserve the first and last 30 bases of each deleted gene.2 www.cdc.gov/biosafety/publications/bmbl5/index.htm. Gene replacement followed a modified Lambda-Red technique, with an added T7 RNA polymerase promoter Disclaimers: positioned in plasmid pCLF3 to generate a gene-specific You are authorized to use this product for research use only. transcript from the Salmonella genome directly downstream of It is not intended for human use. each mutant.2-4 Detailed information about each mutant is shown in Table 1. Use of this product is subject to the terms and conditions of the BEI Resources Material Transfer Agreement (MTA). The Note: The strain designation on the plate, strain CDC 6516- MTA is available on our Web site at www.beiresources.org. 60, is incorrect. The correct strain designation is strain 14028s. S. enterica subsp. enterica, strain 14028s was originally While BEI Resources uses reasonable efforts to include known as strain 14028. A variant of the original strain with a accurate and up-to-date information on this product sheet, rough colony morphology was designated 14028r and the neither ATCC® nor the U.S. Government makes any original smooth strain was renamed 14028s. Strain 14028 is warranties or representations as to its accuracy. Citations a descendent of strain CDC 6516-60 which was isolated from from scientific literature and patents are provided for pools of hearts and livers of 4-week-old chickens.5 The informational purposes only. Neither ATCC® nor the U.S. complete genome of S. enterica subsp. enterica, strain Government warrants that such information has been 14028s (GenBank: CP001363.1) and plasmid confirmed to be accurate. (GenBank: CP001362.1) sequences are available. This product is sent with the condition that you are responsible

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Product Information Sheet for NR-42892

for its safe storage, handling, use and disposal. ATCC® and 2. Porwollik, S., et al. “Defined Single-Gene and Multi-Gene the U.S. Government are not liable for any damages or injuries Deletion Mutant Collections in Salmonella enterica sv arising from receipt and/or use of this product. While Typhimurium.” PLoS One 9 (2014): e99820. PubMed: reasonable effort is made to ensure authenticity and reliability 25007190. of materials on deposit, the U.S. Government, ATCC®, their 3. Santiviago, C. A., et al. “Analysis of Pools of Targeted suppliers and contributors to BEI Resources are not liable for Salmonella Deletion Mutants Identifies Novel Genes damages arising from the misidentification or Affecting Fitness during Competitive Infection in Mice.” misrepresentation of products. PLoS Pathog. 5 (2009): e1000477. PubMed: 19578432. 4. Datsenko, K. A. and B. L. Wanner. “One-step Inactivation Use Restrictions: of Chromosomal Genes in Escherichia coli K-13 Using This material is distributed for internal research, non- PCR Products.” Proc. Natl. Acad. Sci. USA 97 (2000): commercial purposes only. This material, its product or its 6640-6645. PubMed: 10829079. derivatives may not be distributed to third parties. Except as 5. Jarvik, T., et al. “Short-Term Signatures of Evolutionary performed under a U.S. Government contract, individuals Change in the Salmonella enterica Serovar Typhimurium contemplating commercial use of the material, its products or 14028 Genome.” J. Bacteriol. 192 (2010): 560- its derivatives must contact the contributor to determine if a 567. PubMed: 19897643. license is required. U.S. Government contractors may need a license before first commercial sale. ATCC® is a trademark of the American Type Culture Collection. References: 1. McClelland, M., Personal Communication.

Table 1: S. enterica subsp. enterica, Strain 14028s (Serovar Typhimurium) Single-Gene Deletion Mutant Library, Plate SGD_160/161_Cm1,2 Deleted Deleted Well Gene Gene Gene Target Gene Gene Region Region Description Position Type Start End (Locus Tag) Strand Start End A03 CDS 3287807 3288859 STM14_3755 3287837 3288828 - Adenine DNA glycosylase A04 CDS 3276061 3277008 STM14_3739 3276091 3276977 - Glutathione synthetase A05 CDS 3272461 3273855 STM14_3735 3272491 3273824 - Galactose/proton symporter A07 CDS 3270459 3270758 STM14_3732 3270315 3270472 + Putative inner membrane protein A08 CDS 3268048 3270024 STM14_3729 3268078 3269993 + Arginine decarboxylase A09 CDS 3258658 3259578 STM14_3720 3258688 3259547 + Agmatinase A10 CDS 3242418 3243650 STM14_3699 3242448 3243619 + D-3-phosphoglycerate dehydrogenase A11 CDS 3231276 3234149 STM14_3687 3231306 3234229 + Glycine dehydrogenase B02 CDS 3221088 3222821 STM14_3674 3221118 3222790 + ssDNA exonuclease RecJ B05 CDS 3205849 3206361 STM14_3654 3205879 3206330 + Hypothetical protein B06 CDS 3197383 3198144 STM14_3643 3197413 3198113 + 2-deoxy-D-gluconate 3-dehydrogenase B07 CDS 3195611 3197029 STM14_3641 3195641 3196998 + L-arabinose/proton symport protein B08 CDS 3192504 3193766 STM14_3638 3192534 3193735 + Diaminopimelate decarboxylase B10 CDS 3183190 3183885 STM14_3627 3183214 3183854 - DNA mismatch repair protein B11 CDS 3182858 3183004 STM14_3624 3182888 3182974 + Hypothetical protein B12 CDS 3162651 3163982 STM14_3607 3162681 3163951 - N-acetylglutamate synthase C01 CDS 3156970 3157413 STM14_3599 3157000 3157382 - Putative FeS center assembly protein C02 CDS 3141929 3143296 STM14_3582 3141959 3143265 - L-serine dehydratase/L-threonine deaminase 2 C03 CDS 3140581 3141870 STM14_3581 3140611 3141839 - Putative serine transport protein C04 CDS 3133047 3134405 STM14_3570 3133077 3134374 + Putative D-glucarate permease C05 CDS 3130349 3131689 STM14_3568 3130379 3131658 + d-glucarate dehydratase C07 CDS 3113110 3113472 STM14_3554 3113140 3113441 - Putative 6-pyruvoyl tetrahydrobiopterin synthase C08 CDS 3110887 3112686 STM14_3553 3110917 3112655 + Sulfite reductase subunit alpha C09 CDS 3108365 3109099 STM14_3551 3108395 3109068 + Phosphoadenosine phosphosulfate reductase C10 CDS 3094628 3095536 STM14_3539 3094658 3095505 + Sulfate adenylyltransferase subunit 2 C11 CDS 3092587 3093192 STM14_3537 3092617 3093161 + Adenylylsulfate kinase D02 CDS 3022711 3023721 STM14_3454 3022741 3023690 - Putative hydrogenase formation protein D03 CDS 3021593 3022714 STM14_3453 3021623 3022683 - Putative hydrogenase formation protein D04 CDS 3020458 3021330 STM14_3451 3020488 3021299 - Hydrogenase nickel incorporation protein HypB D05 CDS 3018608 3019216 STM14_3446 3018638 3019185 + Hydrogenase-3 iron-sulfur subunit D06 CDS 3016782 3018608 STM14_3445 3016812 3018577 + Formate hydrogenlyase subunit 3 D08 CDS 3014129 3015838 STM14_3443 3014159 3015807 + Hydrogenase 3 large subunit D09 CDS 3012810 3013577 STM14_3441 3012840 3013546 + Hydrogenase D11 CDS 2999103 3000074 STM14_3423 2999133 3000043 - Glucitol/sorbitol-specific enzyme IIB component

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Product Information Sheet for NR-42891

Deleted Deleted Well Gene Gene Gene Target Gene Gene Region Region Description Position Type Start End (Locus Tag) Strand Start End D12 CDS 2998543 2999106 STM14_3422 2998573 2999075 - Glucitol/sorbitol-specific enzyme IIC component E01 CDS 2994472 2994972 STM14_3416 2994502 2994980 + Recombination regulator RecX E02 CDS 2987170 2988726 STM14_3403 2987200 2988695 + Glutamate--cysteine ligase E03 CDS 2977381 2978445 STM14_3392 2977411 2978414 - Glycine betaine transporter membrane protein E04 CDS 2976186 2977388 STM14_3391 2976216 2977357 - Glycine betaine transporter ATP-binding subunit E05 CDS 2972717 2974861 STM14_3389 2972747 2974830 - Ribonucleotide-diphosphate reductase subunit alpha E06 CDS 2972334 2972744 STM14_3388 2972364 2972713 - Ribonucleotide reductase stimulatory protein E07 CDS 2972092 2972337 STM14_3387 2972107 2972306 - Glutaredoxin-like protein E08 CDS 2971817 2971993 STM14_3386 2971847 2971962 - Hypothetical protein E09 CDS 2963245 2964645 STM14_3369 2963275 2964614 - Gamma-aminobutyrate transporter E10 CDS 2961832 2963115 STM14_3368 2961862 2963084 - 4-aminobutyrate aminotransferase E11 CDS 2960369 2961817 STM14_3367 2960399 2961786 - Succinate-semialdehyde dehydrogenase I E12 CDS 2898394 2899458 STM14_3302 2898424 2899427 - Integrase-like protein F03 CDS 2861933 2862097 STM14_3267 2861963 2862066 - Hypothetical protein F04 CDS 2856033 2858606 STM14_3261 2856063 2858575 + Protein disaggregation chaperone F05 CDS 2848209 2849510 STM14_3252 2848239 2849479 + Alpha-ketoglutarate transporter F06 CDS 2842488 2842907 STM14_3246 2842518 2842876 - Thioredoxin 2 F07 CDS 2840442 2841131 STM14_3244 2840472 2841100 - Uracil-DNA glycosylase F08 CDS 2831148 2831627 STM14_3231 2831178 2831596 + SoxR reducing system protein RseC F09 CDS 2829421 2830650 STM14_3228 2829451 2830619 + Integrase-like protein F10 CDS 2829159 2829443 STM14_3227 2829189 2829412 - Excisionase-like protein F11 CDS 2815110 2815250 STM14_3204 2815140 2815219 + Hypothetical protein F12 CDS 2809856 2810257 STM14_3193 2809886 2810226 - Hypothetical protein G01 CDS 2809856 2810257 STM14_3193 2809886 2810226 - Hypothetical protein G02 CDS 2809024 2809569 STM14_3192 2809054 2809538 + DNA packaging-like protein G04 CDS 1308154 1309773 STM14_1458 1308223 1309742 - Phage portal protein, lambda family G06 CDS 1320754 1321455 STM14_1472 1320787 1321424 - Phage tail component L-like protein G08 CDS 2774738 2775466 STM14_3159 2774768 2775435 + DNA repair protein RecO G09 CDS 2773995 2774726 STM14_3158 2774025 2774695 + Pyridoxal phosphate biosynthetic protein PdxJ G10 CDS 2759807 2763694 STM14_3145 2759837 2763783 + Phosphoribosylformylglycinamidine synthase G11 CDS 2751503 2753647 STM14_3138 2751533 2753616 - Lysine decarboxylase 1 H05 CDS 2729749 2731599 STM14_3114 2729779 2731568 + Chaperone protein HscA H06 CDS 2729412 2729747 STM14_3113 2729442 2729716 + Electron carrer protein H07 CDS 2727670 2728953 STM14_3111 2727700 2728922 + Aminopeptidase B H08 CDS 2726784 2727569 STM14_3110 2726814 2727538 + Enhanced serine sensitivity protein SseB H10 CDS 2677631 2678980 STM14_3077 2677661 2678949 - Exodeoxyribonuclease VII large subunit 1All information in this table was provided by the depositor at the time of deposition. 2Construction of each listed mutant has been confirmed either by PCR or by an array indicating a functional T7 promoter in the correct location and orientation. Mutants that did not produce such a signal on the array, or did not yield the expected mutant product during PCR, are not listed.

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