Abstract
We report the isolation of a group of 279 Salmonella typhimurium strains carrying randomly spaced insertions of the minitransposon Tn10 delta 16 delta 17 and describe the use of these strains to facilitate genetic analysis. The insertions were isolated initially in individual recombinant lambda clones from a genomic library. Individual insertions were then moved into the S. typhimurium chromosome, where the distribution of insertion sites relative to standard genetic markers was analyzed in a series of transductional crosses. Since a different, randomly chosen clone was used to generate each insertion, the distribution of insertion positions should have been as random as the cloning events leading to the formation of the library. In agreement with this expectation, most S. typhimurium markers tested were cotransducible with one or more of these Tn10 delta 16 delta 17 insertions. We expect that most new mutations will be quickly classified and mapped by determination of the pattern of cotransduction with this set of insertions. This use is illustrated by the analysis of a group of lac operon fusions regulated by anaerobiosis. We also describe several other applications that should make this collection a useful new tool in S. typhimurium genetics.
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Chumley F. G., Menzel R., Roth J. R. Hfr formation directed by tn10. Genetics. 1979 Apr;91(4):639–655. doi: 10.1093/genetics/91.4.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Engstrom J., Wong A., Maurer R. Interaction of DNA polymerase III gamma and beta subunits in vivo in Salmonella typhimurium. Genetics. 1986 Jul;113(3):499–515. doi: 10.1093/genetics/113.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Enomoto M., Stocker B. A. Transduction by phage P1kc in Salmonella typhimurium. Virology. 1974 Aug;60(2):503–514. doi: 10.1016/0042-6822(74)90344-4. [DOI] [PubMed] [Google Scholar]
- Foster T. J., Davis M. A., Roberts D. E., Takeshita K., Kleckner N. Genetic organization of transposon Tn10. Cell. 1981 Jan;23(1):201–213. doi: 10.1016/0092-8674(81)90285-3. [DOI] [PubMed] [Google Scholar]
- Halling S. M., Kleckner N. A symmetrical six-base-pair target site sequence determines Tn10 insertion specificity. Cell. 1982 Jan;28(1):155–163. doi: 10.1016/0092-8674(82)90385-3. [DOI] [PubMed] [Google Scholar]
- Hmiel S. P., Snavely M. D., Miller C. G., Maguire M. E. Magnesium transport in Salmonella typhimurium: characterization of magnesium influx and cloning of a transport gene. J Bacteriol. 1986 Dec;168(3):1444–1450. doi: 10.1128/jb.168.3.1444-1450.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hohn B. In vitro packaging of lambda and cosmid DNA. Methods Enzymol. 1979;68:299–309. doi: 10.1016/0076-6879(79)68021-7. [DOI] [PubMed] [Google Scholar]
- Karn J., Brenner S., Barnett L., Cesareni G. Novel bacteriophage lambda cloning vector. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5172–5176. doi: 10.1073/pnas.77.9.5172. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kleckner N., Roth J., Botstein D. Genetic engineering in vivo using translocatable drug-resistance elements. New methods in bacterial genetics. J Mol Biol. 1977 Oct 15;116(1):125–159. doi: 10.1016/0022-2836(77)90123-1. [DOI] [PubMed] [Google Scholar]
- Maurer R., Osmond B. C., Shekhtman E., Wong A., Botstein D. Functional interchangeability of DNA replication genes in Salmonella typhimurium and Escherichia coli demonstrated by a general complementation procedure. Genetics. 1984 Sep;108(1):1–23. doi: 10.1093/genetics/108.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maurizi M. R., Trisler P., Gottesman S. Insertional mutagenesis of the lon gene in Escherichia coli: lon is dispensable. J Bacteriol. 1985 Dec;164(3):1124–1135. doi: 10.1128/jb.164.3.1124-1135.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palva E. T., Liljeström P., Harayama S. Cosmid cloning and transposon mutagenesis in Salmonella typhimurium using phage lambda vehicles. Mol Gen Genet. 1981;181(2):153–157. doi: 10.1007/BF00268420. [DOI] [PubMed] [Google Scholar]
- Sanderson K. E., Roth J. R. Linkage map of Salmonella typhimurium, Edition VI. Microbiol Rev. 1983 Sep;47(3):410–453. doi: 10.1128/mr.47.3.410-453.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strauch K. L., Lenk J. B., Gamble B. L., Miller C. G. Oxygen regulation in Salmonella typhimurium. J Bacteriol. 1985 Feb;161(2):673–680. doi: 10.1128/jb.161.2.673-680.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wasserman S. A., Walsh C. T., Botstein D. Two alanine racemase genes in Salmonella typhimurium that differ in structure and function. J Bacteriol. 1983 Mar;153(3):1439–1450. doi: 10.1128/jb.153.3.1439-1450.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Vries G. E., Raymond C. K., Ludwig R. A. Extension of bacteriophage lambda host range: selection, cloning, and characterization of a constitutive lambda receptor gene. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6080–6084. doi: 10.1073/pnas.81.19.6080. [DOI] [PMC free article] [PubMed] [Google Scholar]