Summary
Secondary attachment site λ-lysogens were isolated in an Escherichia coli strain carrying multiple tandem 9.8 kb repeats. The repeat carried the structural gene for chromosomal β-lactamase, ampC. One lysogen produced lysates with amp-transducing activity. Three types of phages with different densities were obtained from this lysogen. The one with the lowest density was found to be a helper λcI857S7 phage. The other two phages showed identical restriction endonuclease fragmentation patterns. The difference in density was due to the presence or absence of phage tail. In λdamp the right cohesive end segment was deleted in a random fashion with the majority ending between 81.0% and 82.4% of λ. The chromosomal segment of λdamp was most likely located at the λ attachment site. The λdamp DNA was compared to that of a ColE1 hybrid carrying the chromosomal amp segment and a ColE1 hybrid carrying the same 9.8 kb amp repeat as the lysogen from which λdamp was isolated. It was found that the chromosomal part of λdamp constituted 9.8 kb, i.e. the size of one repeat. Moreover, the novel joint between adjacent repeats was present. In a λattB-deleted E. coli K-12 strain, lysogenic for λdamp, highly ampicillin-resistant mutants occurred at an exceedingly high frequency. They were found to contain in the chromosome an amplified 9.8 kb repeat. This suggested that integration of the novel joint from λdamp into the amp region gives rise to an amplifiable duplication. In E. coli lysogenized for λdamp at λattB highly ampicillin-resistant clones were also found at a high frequency. These clones carried multiple tandem repeats of λdamp DNA, each with an intact right end segment.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andersson RP, Roth JK (1977) Tandem genetic duplication in phage and bacteria. Annu Rev Microbiol 31:473–505
Andersson RP, Roth JR (1978) Tandem genetic duplications in Salmonella typhimurium: Amplification of the histidine operon. J Mol Biol 126:53–71
Andersson RP, Miller CG, Roth JR (1976) Tandem duplication of the histidine operon observed following generalized transduction in Salmonella typhimurium. J Mol Biol 105:201–210
Bachmann BJ, Low BK (1980) Linkage map of Escherichia coli K-12. Microb Rev 44:1–56
Bertani G (1951) Studies on lysogenesis I. The mode of phageliberation by lysogenic Escherichia coli. J Bacteriol 62:293–300
Burman LG, Park JT, Lindström EB, Boman HG (1973) Resistance of Escherichia coli to penicillinase X. Identification of the structural gene for the chromosomal penicillinase. J Bacteriol 116:123–130
Edlund T, Grundström T, Normark S (1979) Isolation and characterization of DNA repetitions carrying the chromosomal β-lactamase gene of Escherichia coli K-12. Mol Gen Genet 173:115–125
Feiss M, Fisher RA, Crayton MA, Egrer C (1977) Packaging of the bacteriophage lambda chromosome: effect on chromosome length. Virology 77:281–293
Hill CW, Combriato G (1973) Genetic duplications induced at a very high frequency by ultraviolet irradiation in Escherichia coli. Mol Gen Genet 127:197–214
Hohn T, Katsura I (1977) Structure and assembly of bacteriophage lambda. Curr Top Microbiol Immunol 78:69–110
Katsura J (1976) Morphogenesis of bacteriophage lambda tail, polymorphism in the assembly of the major tail protein. J Mol Biol 107:307–326
Langridge J (1969) Mutations conferring quantitative and qualitative increases in β-galactosidase activity in Escherichia coli. Mol Gen Genet 105:74–83
Miller JH (1974) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor New York
Normark S (1970) Genetics of a chain-forming mutant of Escherichia coli. Transduction and dominance of the envA gene mediating increased penetration to some antibacterial agents. Genet Res 16:63–78
Normark S, Burman LG (1977) Resistance of Escherichia coli to penicillins: fine mapping and dominance of chromosomal β-lactamase mutations. J Bacteriol 132:1–7
Normark S, Edlund T, Grundström T, Bergström S, Wolf-Watz H (1977) Escherichia coli K-12 mutants hyperproducing chromosomal β-lactamase by gene repetitions. J Bacteriol 132:912–922
Schrenk WJ, Weisberg RA (1975) A simple method for making new transducing lines of coliphage λ. Mol Gen Genet 137:101–107
Shimada K, Weisberg R, Gottesman ME (1973) Prophage lambda at unusual chromosomal locations. II Mutations induced by bacteriophage lambda in Escherichia coli K-12. J Mol Biol 80:297–314
Smith GP (1973) Unequal crossover and the evolution of multigene families. Cold Spring Harbor Symp Quant Biol 38:507–513
Smith GP (1976) Evolution of repeated DNA by unequal crossover. Science 191:528–535
Sternberg N, Weisberg R (1975) Packaging of prophage and host DNA by coliphage λ. Nature 256:97–103
Straus DS (1974) Induction by mutagenesis of tandem gene duplications in the glyS region of the Escherichia coli chromosome. Genetics 78:823–830
Straus DS, Straus HD (1976) Large overlapping tandem genetic duplications in Salmonella typhimurium. J Mol Biol 103:143–154
Sutcliffe JG (1978) pBR322 restriction map derived from the DNA sequence: Accurate DNA size markers up to 4361 nucleoside pairs long. Nucleic Acid Res 5:2721–2728
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Edlund, T., Grundström, T., Björk, G.R. et al. Tandem duplication induced by an unusual ampA1-, ampC-transducing lambda phage: A probe to initiate gene amplification. Molec. Gen. Genet. 180, 249–257 (1980). https://doi.org/10.1007/BF00425836
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00425836