Summary
Nocardia mediterranei strain LBG A3136 contains the 23.7 kb element pMEA100 in a chromosomally integrated form as well as in the free state (Moretti et al. 1985). The integrated form of this element can be excised precisely from the Nocardia chromosome without any accompanying rearrangements in flanking chromosomal DNA. After transfer into plasmid-free mutant strains, pMEA100 reintegrates site specifically into its original chromosomal locus. The exact mapping of the pMEA100 integration site was accomplished by restriction analysis and DNA sequencing. The attachment site of pMEA100, the junctions of its integrated form and plasmid-free chromosomal DNA of N. mediterranei contain an identical 47 bp long sequence which is probably required for site-specific recombination connected with integration and excision of pMEA100. Only one such sequence was found in the chromosome of pMEA100-free N. mediterranei derivatives as suggested by the single integration locus.
Similar content being viewed by others
References
Andrews BJ, Proteau GA, Beatty LG, Sadowski PD (1985) The FLP recombinase of the 2 μ circle DNA of yeast: interaction with its target sequences. Cell 40:795–803
Bibb MJ, Ward JM, Kieser T, Cohen SN, Hopwood DA (1981) Excision of chromosomal DNA sequences from Streptomyces coelicolor forms a novel family of plasmids detectable in Streptomyces lividans. Mol Gen Genet 184:230–240
Birner J, Hodgson PR, Lane WR, Baxter EH (1972) An Australian isolate of Nocardia mediterranea producing rifamycin SV. J Antibiot 25:356–359
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113
Cohen A, Bar-Nir D, Goedeke ME, Parag Y (1985) The integrated and free states of Streptomyces griseus plasmid pSG1. Plasmid 13:41–50
Frischauf A-M, Lehrach H, Poustka A, Murray N (1983) Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170:827–842
Ghisalba O, Auden JAL, Schupp T, Nüesch J (1984) The rifamycins: properties, biosynthesis and fermentation. In: Vandamme EJ (ed) Biotechnology of industrial antibiotics. Dekker, New York Basel, p 281
Hoess RH, Abremski K (1984) Interaction of the bacteriophage P1 recombinase Cre with the recombining site loxP. Proc Natl Acad Sci USA 81:1026–1029
Hohn B, Hinnen A (1980) Cloning with cosmids in E. coli and yeast. In: Setlow JK, Hollaender A (eds) Genetic engineering. Principles and methods 2. Plenum Press, New York, p 169
Hopwood DA, Hintermann G, Kieser T, Wright HM (1984) Integrated DNA sequences in three streptomycetes form related autonomous plasmids after transfer to Streptomyces lividans. Plasmid 11:1–16
Hsu P-L, Ross W, Landy A (1980) The λ phage att site: functional limits and interaction with Int protein. Nature 285:85–91
Kaluza K (1985) Strukturanalyse des nif DK Operons und repetitiver Sequenzen von Rhizobium japonicum. Thesis ETH, Zürich, No 7831
Kieser T (1984) Factors affecting the isolation of cccDNA from Streptomyces lividans and Escherichia coli. Plasmid 12:19–36
Lydiate DJ, Ikeda H, Hopwood DA (1986) A 2.6 kb DNA sequence of Streptomyces coelicolor A3(2) which functions as a transposable element. Mol Gen Genet 203:79–88
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Maxam AM, Gilbert W (1980) Sequencing end-labelled DNA with base specific chemical cleavages. Methods Enzymol 65:499–560
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York
Moretti P, Hintermann G, Hütter R (1985) Isolation and characterization of an extrachromosomal element from Nocardia mediterranei. Plasmid 14:126–133
Omer CA, Cohen SN (1984) Plasmid formation in Streptomyces: excision and integration of the SLP1 replicon at a specific chromosomal site. Mol Gen Genet 196:429–438
Omer CA, Cohen SN (1986) Structural analysis of plasmid and chromosomal loci involved in site-specific excision and integration of the SLP1 element of Streptomyces coelicolor. J Bacteriol 166:999–1006
Pernodet J-L, Simonet J-M, Guerineau M (1984) Plasmids in different strains of Streptomyces ambofaciens: free and integrated form of plasmid pSAM2. Mol Gen Genet 198:35–41
Pridham TG, Anderson P, Foley C, Lindenfelser LA, Hesseltine CW, Benedict RG (1956/1957) A selection of media for maintenance and taxonomic study of Streptomyces. Antibiotics Ann 947–953
Schär M (1984) Klonieren von Nocardia mediterranei DNA in E. coli und Sequenzieren eines Nocardia mediterranei DNA Fragmentes mit komplementierenden Eigenschaften in einer Adenin-Mutante von Streptomyces lividans. Thesis Universität Basel, Basel
Schupp T (1973) Genetik von Nocardia mediterranei. Thesis ETH, Zürich, No 5153
Schupp T, Hütter R, Hopwood DA (1975) Genetic recombination in Nocardia mediterranei. J Bacteriol 121:128–136
Weisberg RA, Landy A (1983) Site-specific recombination in phage lambda. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory, pp 211–250
Author information
Authors and Affiliations
Additional information
Communicated by W. Arber
Rights and permissions
About this article
Cite this article
Madon, J., Moretti, P. & Hütter, R. Site-specific integration and excision of pMEA100 in Nocardia mediterranei . Molec Gen Genet 209, 257–264 (1987). https://doi.org/10.1007/BF00329651
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00329651