SALMONELLA GENETIC STOCK CENTRE (SGSC)

Kits of Strains

Over a period of years "kits" of strains have been developed for solving specific problems in genetic analysis. Most of the materials were generously provided by other investigators for use by the whole community. These kits of strains and/or phages are listed below, click on the kit number or scroll down page to see a brief description. Most of the brief descriptions also have clickable links to more detailed descriptions including strain lists.


Listing of Kits Maintained at the SGSC:

Kit 1. Transposons
Kit 2. Mutants organized by gene or operon
Kit 3. Hfr strains of S. typhimurium and S. abony
Kit 4. F-prime factor with E. coli DNA
Kit 5. Delivery Systems for construction of transposition strains in S. typhimurium
Kit 6. Transducing phages for S. typhimurium
Kit 7. Adding tails to P22
Kit 8. Banks of cloned DNA of Salmonella
Kit 9. Extending the host range of bacteriophages
Kit 10. Displacing pSLT from S. typhimurium
Kit 11. Streptomycin resistant point mutants in S. typhimurium
Kit 12. Strains for homologous recombination of genes transferred in to S. typhimurium
Kit 13. Hybrid plasmids carrying S. typhimurium genes
Kit 14. Wild type strains of Salmonella species


1. Transposon kits

1A. Transposons in the genome of S. typhimurium - Insertions of transposons into and between known genes of S. typhimurium. This includes over 1,000 strains, including insertions of Tn10, MudA, MudJ, MudK, MudP22, and others. Strains were provided by many investigators. (Ref: Altman, E. et. al. 1996. Transposons currently in use in genetic analysis of Salmonella species. In E.coli and Salmonella, 2nd Edn., Ed. Neidhardt, F.C. ASM Press.1996., Volume 2 Chapter 141, Table 2 and Sanderson, K.E. and Roth, J. R. 1983. Microbiol. Rev. 47:410-453).

1B. Transposons used to insert specific mutations into strains - Some strains in this transposon kit carry mutations which are linked by P22 transduction to the transposon such as Tn10 which has a selectable marker; thus this strain can be used to transfer the mutation into new backgrounds. There are many such examples, including strains which can be used to transfer galE or recA alleles.

1C. Random Tn10 insertion mutations in the chromosome - A group of strains with random Tn10 insertions into the chromosome of S. typhimurium LT2 was constructed (Kukral, A. M. et al. 1987. J. Bacteriol. 169:1787-1793.); of these 276 are available from the SGSC. Many of these insertions have been located on the chromosome, and are listed in kit 1A, but about half are not yet located.

1D. The MudP and MudQ (Mud-P22) Mapping Set - Youderian et al. developed a P22 phage with Mu-ends which lysogenizes at many sites but which cannot detach from the chromosome, so is "locked in"; when induced it replicates and packages about 100 to 200 kb of chromosomal DNA in the insertion region. Benson and Goldman developed a set of about 50 strains, each of which have MudP22 inserted at different sites around the chromosome.


2. Kits of mutants organized by gene or operon

2A. Lipopolysaccharide mutants - Set of mutant strains of S. typhimurium LT2, representing defects in the lipopolysaccharide.

2B. Histidine mutants - Sets of mutants of Salmonella typhimurium LT2 affecting the his (histidine) genes. These mutants were obtained from the laboratory of Philip E. Hartman, Dept. of Biology, The John Hopkins University, Baltimore, MD 21218. The following three kits have been constructed:

Kit 2B.1. Point Mutants in Genes of the Histidine Operon

Kit 2B.2. Deletion Mutants in the Histidine Operon

Kit 2B.3. Set of F-Prime Strains with Mutations in the his Genes of E. coli K-12


3. Hfr strains of S. typhimurium and S. abony

3. Hfr strains of S. typhimurium and S. abony - The F-factor of E. coli K-12 was transferred into S. typhimurium and S. abony, and a set of Hfr strains was isolated by Sanderson et al., 1972. Bacteriol. Rev. 36:603-637.


4. F-prime factor with E. coli DNA

4. F-prime factors with E. coli DNA - This kit includes strains of S. typhimurium LT2 which carry F-prime factors which carry genes derived from E. coli K-12, representing a large part (but not all) of the chromosome of E. coli. They were constructed originally by Brooks Low, and transferred into S. typhimurium by conjugation.


5. Delivery Systems for construction of transposition strains in S. typhimurium

5A. Tn5 Delivery Systems:

5A.1. P22 Generalized Transduction of Tn5 - Utilizes P22 to construct insertional mutants with the transposon Tn5. Generalized transduction by P22 delivers a segment of E. coli. DNA containing Tn5 (Kan (R)); Kanamycin resistant transductants in S. typhimurium arise only from the transposition.

5A.2. Delivery System for Lambda-Sensitive Tn5 using Lambda into Salmonella - Use of phage transfer by Lambda to deliver a Lambda::Tn5 (Kan (R)) into Salmonella. Kanamycin resistant recombinants arise only from the transposition.

5B. Tn10 Delivery Systems:

5B. 1. P22 Generalized Transduction of Tn10 - Utilizes P22 to construct insertional mutants with the transposon Tn10. Generalized transduction by P22 delivers a segment of E. coli. DNA containing Tn10 (Tet (R)); Tetracycline resistant transductants in S. typhimurium arise only from the transposition.

5B.2. Pre- Constructed Pools of Tn10 Insertions in S. typhimurium LT2 - pools of S. typhimurium LT2 which were transduced to Tc(R) by P22::Tn10 in the laboratory of J.R. Roth.

5C. Mudlac (MudJ) Delivery Systems - Material provided by John Roth, University of Utah. A complementation method uses P22 to transduce a defective Mu element which has the operon fusion lacZ reporter gene (Hughes, K.T., Roth, J.E. 1988. Genetics 119: 9-12.). It was developed to permit simple isolation of single mutants due to insertion of defective elements.

5D. TnphoA Delivery Systems - Material provided by Ron Taylor. The plasmids pRT291 and pRT733 are used for delivery of TnphoA into Gram-negative bacteria. In S. typhimurium it is necessary to use a Pho(-) mutant, lacking acid phosphatase, as a recipient strain.


6. Transducing phages for S. typhimurium

6A. General transducing phages for S. typhimurium - Wild type forms and (in some cases) mutants of phages which give generalized transduction. Phages available are: P22 and its derivative P22 HT105 int1; ES18 and ES18h1; and P1-Cm cts phage.

6B. LPS Specific phage - The lipopolysaccharide specific phages available are: Felix 'O' and Ffm


7. Kit for adding tails to P22

7. System for Adding Tails to P22


8. Banks of cloned DNA of Salmonella

8A. BAC clones of S. Typhimurium LT2 (SGSC1412) - A set of pBeloBACII clones in E. coli that represent almost the entire S. typhimuium LT2 (SGSC1412) genome and pSLT. Clones were used by the Washington University Genome Sequencing Center for end-sequencing and generation of STS markers (Ref: McClelland M, Sanderson KE, Spieth J, Clifton SW, Latreille P, Courtney L, Porwollik S, Ali J, Dante M, Du F, Hou S, Layman D, Leonard S, Nguyen C, Scott K, Holmes A, Grewal N, Mulvaney E, Ryan E, Sun H, Florea L, Miller W, Stoneking T, Nhan M, Waterston R, Wilson RK. Complete genome sequence of Salmonella enterica serovar Typhimurium LT2. Nature. 2001 Oct 25;413(6858):852-6.)

8B. Sau3A library, packaged in P22 - A library of pBR322 derivatives was generated by ligating partial Sau3A digests of wild type S. typhimurium LT2 into the BamHI site of the vector, yielding about 15 kb inserts. This was transformed into S. typhimurium LT2 LB5000, and cells are stored in glycerol at low temperature; cells are available on request. In addition, a P22 lysate was produced on these cells, and this lysate can be used to transfer hybrid plasmids into cells by transduction (Source N. M. Kredich; Monroe, R. S. and N. M. Kredich. 1988. J. Bacteriol. 170:42-47)

8C. Lambda 1059 library - S. typhimurium DNA was Sau3A digested and ligated into Lambda 1059, packaged in vitro, transfected into E. coli cells, and amplified as five separate pools (Constructed by Russ Maurer, following the procedures of Karn et al.,1980).

8D. Partially Ordered Set of Lambda Clones Containing Inserts of Salmonella typhimurium DNA - As part of the ongoing sequencing of the complete Salmonella typhimurium LT2 genome, a partly ordered set of 416 lambda clones has been developed, representing over 90% of the genome (Wong et al .,1999).


9. Systems to extend the host range of bacteriophages

9A. Systems to extend the host range of phage lambda - This plasmid expresses the lamB gene of E. coli K-12; the Lambda receptor protein is displayed on the cell surface (Source: E. T. Palva, Helsinki, Finland; Harkki, A. and E. T. Palva. 1985. FEMS Microbiol. Letters 27:183-187).

9B. Transduction in E. coli with Salmonella phage P22 - The laboratory of Peter Reeves has constructed a system which allows the propagation of P22 in E. coli. This allows the use of P22 for transduction in E. coli, and potentially in other systems (Neal et al. 1993. J. Bacteriol 175 (21):7115-7118).


10. Kit for displacement of pSLT from S. typhimurium

10. Displacement of pSLT from S. typhimurium - The 90 kb resident of S. typhimurium (pSLT, also called virulence plasmid) replicates very stably and is invariably found in lines of LT2 unless it has been intentionally eliminated. A plasmid, pLL6, was constructed which permits displacement and curing of pSLT (R. Kelln, and Lintott LG.1990. Mol. Gen. Genet. 222:438-440).


11. Kit of streptomycin resistant point mutants in S. typhimurium

11. Streptomycin resistant point mutants in S. typhimurium - A set of about 40 streptomycin resistant strains, each with a single auxotrophic mutation at known locations around the chromosome, has been constructed and is available.


12. Strains for homologous recombination of genes transferred in to S. typhimurium

12. Strains for homologous recombination of genes transferred into S. typhimurium - Strains of S. typhimurium with mutS, mutL, or mutH mutations, defective in mismatch repair, give greatly increased frequency of homologous recombination. This permits transfer of chromosome segments including mutant alleles in genes in E. coli or other genera into S. typhimurium by conjugation or transduction. These mut strains are also host-restriction-deficient (Rayssiguier, C., et al. 1989. Nature 342: 396-401).


13. Hybrid plasmids carrying S. typhimurium genes

13. A partial set of cloned genes of S. typhimurium - Plasmids (multi-copy and mostly ColE1-type) assembled from many sources, using a variety of cloning vehicles, which have been assembled as a set to permit probing of portions of the chromosome.


14. Sets of wild type strains of Salmonella species

14A. SAlmonella Reference Collection A (SARA) - A set of 72 strains of Salmonella collected from a variety of sources representing strains from the part of Subgenus I which includes S. typhimurium. These are separated into 48 electrophoretic types using Multi-Locus Enzyme Electrophoresis (MLEE) (Beltran, P. et al., 1991. J. Gen. Microbiol. 137:601-606.).

14B. SAlmonella Reference Collection B (SARB) - A set of Salmonella strains collected from a variety of sources, representing 37 serovars of Subspecies I. These are separated into electrophoretic types using Multi-Locus Enzyme Electrophoresis (MLEE) (Boyd et al., 1993. J. Gen. Microbiol. 139: 1125-1132.).

14C. SAlmonella Reference Collection C (SARC) - A set of Salmonella strains collected from a variety of sources, representing all eight subgenera, including Subgenus V (called S. bongori). These are separated into electrophoretic types using Multi-Locus Enzyme Electrophoresis (MLEE) (Boyd et al, 1996. Appl. and Envir. Microbiol. 62(3):804-808).

14D. Lilleengen Type (LT) strains - A large number of Salmonella typhimurium strains was classified according to sensitivities to bacteriophages. From this, 22 different type strains were classified and one strain of each type was selected and named (Lilleengen, K. 1948. Acta. Pathol. Microbiol. Scand. Suppl. 77: 11-125). The 22 strains collected by Lilleengen were analyzed by Zinder and Lederberg, and lyophils from early transfers were provided by J. Lederberg to the SGSC.


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File updated April 15, 2002