Summary
We present a method that allows positive selection and rapid analysis of mutations in Enterobacteriaceae. Mutations are detected in a 2630 bp selection cartridge inserted in two different bacterial mutlicopy plasmid vectors. Spontaneous mutations in Escherichia coli, Enterobacter cloacae and Citrobacter freundii include insertions, deletions and point mutations. The small size of the target sequence facilitates rapid analysis of DNA rearrangements by cleavage with restriction enzymes and of any type of mutation by DNA sequence analysis. While in E. coli insertions of the mobile elements IS1, IS2 and IS5 were readily found, insertions of putative new transposable elements were detected in Enterobacter cloacae. The selection cartridge can thus serve as a tool for studying the spectrum of insertion mutations in Enterobacteriaceae and probably other Gramnegative bacteria, and the dependency of this spectrum on physiological and environmental factors and the host's genetic background can be investigated.
References
Arber W, Iida S, Jütte H, Caspers P, Meyer J, Hänni C (1978) Rearrangements of genetic material in Escherichia coli as observed on the bacteriophage P1 plasmid. Cold Spring Harbor Symp Quant Biol 43:1197–1208
Bagdasarian M, Timmis KN (1982) Host vector systems for gene cloning in Pseudomonas. Curr Top Microbiol Immunol 96:47–67
Beck E, Ludwig G, Auerswald EA, Reiss B, Schaller H (1982) Nucleotide sequence and exact localisation of the neomycin phosphotransferase gene from transposon Tn5. Gene 19:327–336
Berg DE, Berg CM (1983) The prokaryotic transposable element Tn5. Biotechnology 1:417–435
Bishop R, Sherratt D (1984) Transposon Tn1 intramolecular transposition. Mol Gen Genet 196:117–122
Chen EJ, Seeburg PH (1985) Supercoil sequencing: a fast simple method for sequencing plasmid DNA. DNA 4:165–170
Daniels DL, Schroeder JL, Szybalski W, Sanger F, Coulson AR, Hong GF, Hill DF, Petersen GB, Blattner FR (1983) Complete annotated lambda sequence. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 519–676
Flashman SM (1978) Mutational analysis of the operators of bacteriophage lambda. Mol Gen Genet 166:61–73
Follath F, Costa E, Thommen A, Frei R, Burdeska A, Meyer J (1987) Clinical consequences of development of resistance to third generation cephalosporins. Eur J Clin Microbiol 6:446–450
Gay P, Lecoq D, Steinmetz M, Berkelman T, Kado CI (1985) Positive selection procedure for entrapment of insertion sequence elements in gram-negative bacteria. J Bacteriol 164:918–921
Ghosal D, Saedler H (1977) Isolation of the mini insertions IS6 and IS7 of E. coli. Mol Gen Genet 158:123–128
Ghosal D, Sommer H, Saedler H (1979) Nucleotide sequence of the transposable DNA-element IS2. Nucleic Acids Res 6:1111–1122
Gussin GN, Johnson DA, Pabo CO, Sauer RT (1983) Repressor and cro protein: Structure, function and role in lysogenization. In: Hendrix RW, Roberts JW, Stahl EW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 93–121
Iida S, Meyer J, Kennedy KE, Arber W (1982) A site specific conservative recombination system carried by bacteriophage P1. Mapping the recombinase gene cin and the crossover sites cix for the inversion of the C segment. EMBO J 1:1445–1453
Iida S, Meyer J, Arber W (1983) Prokaryotic IS elements. In: Shapiro JA (ed) Mobile Genetic Elements. Academic Press, New York, pp 159–221
Iida S, Kulka I, Meyer J, Arber W (1987) Amplification of drug resistance genes flanked by inversely repeated IS1 elements: Involvement of IS1-promoted DNA rearrangements before amplification. J Bacteriol 169:1447–1453
Jaurin B, Normark S (1983) Insertion of IS2 creates a novel ampC promoter in Escherichia coli. Cell 32:809–816
Johnsrud L (1979) DNA sequence of the transposable element IS1. Mol Gen Genet 169:213–218
Kröger M, Hobom G (1982) Structural analysis of insertion sequence IS5. Nature 297:159–162
Lucchesi P, Carraway M, Marinus MG (1986) Analysis of forward mutations induced by N-methyl-N′-nitro-N-nitrosoguanidine in the bacteriophage P22 mnt repressor gene. J Bacteriol 166:34–37
Luria SE, Delbrück M (1943) Mutations of bacteria from virus sensitivity to virus resistance. Genetics 28:491–511
Messing J (1979) A multi-purpose cloning system based on the single-stranded DNA bacteriophage M13. Recomb DNA Tech Bull 2:43–48
Meyer BJ, Maurer R, Ptashne M (1980) Gene regulation at the right operator (O R) of bacteriophage λ. II. O R1, O R2, and O R3: Their roles in mediating the effect of repressor and cro. J Mol Biol 139:163–194
Miller JH (1983) Mutational specificity in bacteria. Annu Rev Genet 17:215–238
Rak B, van Reutern M (1984) Insertion element IS5 contains a third gene. EMBO J 3:807–811
Roberts TM, Schwanberg SL, Poteete A, Riedel G, Backman K (1980) A plasmid cloning vehicle allowing a positive selection for inserted fragments. Gene 12:123–127
Rothstein SJ, Reznikoff WS (1981) The functional differences in the inverted repeats of Tn5 are caused by a single base pair nonhomology. Cell 23:191–199
Schaaper RM, Danforth BN, Glyckman BW (1985) Rapid repeated cloning of mutant lac repressor genes. Gene 39:181–189
Schaaper RM, Danforth BN, Glyckman BW (1986) Mechanisms of spontaneous mutagenesis: an analysis of the spectrum of spontaneous mutation in the Escherichia coli lacI gene. J Mol Biol 189:273–284
Schoner B, Kahn M (1981) The nucleotide sequence of IS5 from Escherichia coli. Gene 14:165–174
Sengstag C, Arber W (1983) IS2 insertion is a major cause of spontaneous mutagenesis of the bacteriophage P1: non-random distribution of target sites. EMBO J 2:67–71
Sengstag C, Arber W (1987) A cloned DNA fragment from bacteriophage P1 enhances IS2 insertion. M l Gen Genet 206:344–351
Stüber D, Bujard H (1981) Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci USA 78:167–171
Sutcliffe JG (1979) Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43:77–90
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
Wood WB (1966) Host specificity of DNA produced by Escherichia coli: Bacterial mutations affecting the restriction and modification of DNA. J Mol Biol 16:118–133
Youderian P, Vershov A, Bouvier S, Sauer RF, Susskind MM (1983) Changing the DNA-binding specificity of a repressor. Cell 35:777–783
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Communicated by H. Hennecke
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Raabe, T., Jenny, E. & Meyer, J. A selection cartridge for rapid detection and analysis of spontaneous mutations including insertions of transposable elements in Enterobacteriaceae. Mol Gen Genet 215, 176–180 (1988). https://doi.org/10.1007/BF00331322
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DOI: https://doi.org/10.1007/BF00331322