Summary
The maintenance and genetic stability of the vector plasmids pBR322 and pBR325 in two genetically different Escherichia coli hosts were studied during chemostat cultivation with glucose and ammonium chloride limitation and at two different dilution rates. The plasmid pBR322 was stably maintained under all growth conditions tested. However pBR325 segregated from both hosts preferentially during glucose limitation and at low dilution rate. In addition to this general segregation process a separate loss of tetracycline resistance was observed. The remaining plasmid conferred resistance to ampicillin and chloramphenicol only, without any remarkable alteration of its molecular weight.
Cultivation conditions in the chemostat were found that allowed the stable genetic inheritance of both plasmids in the hosts studied.
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References
Adams J, Kinney T, Thompson S, Rubin L, Helling RB (1979) Frequency-dependent selection for plasmid-containing cells of E. coli. Genetics 91:627–637
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid. Nucleic Acids Res 7:1513–1523
Bolivar F, Rodriguez RL, Greene PJ, Betlach MO, Heyneker HL, Boyer HW, Crosa IH, Falkow S (1977) Construction and characterization of new cloning vehicles; II. A multiple cloning system. Gene 2:95–113
Bolivar F (1978) Construction and characterization of new cloning vehicles; III. Derivatives of plasmid pBR322 carrying EcoR1 sites for selection of EcoR1 generated recombinant DNA molecules. Gene 4:121–136
Dagert M, Ehrlich SD (1979) Prolonged incubation in CaCl2 improves the competence of E. coli cells. Gene 6:23–28
Godwin D, Slater JH (1979) The influence of the growth environment on the stability of a drug resistance plasmid in E. coli K12. J Gen Microbiol 111:201–210
Hashimoto-Gotoh T, Inselburg J (1979) Isolation and characterization of replication-deficient mutants of ColE1 plasmids. J Bacteriol 139:597–607
Herbert D, Elsworth R, Telling RC (1956) The continuous culture of bacteria: a theoretical and experimental study. J Gen Microbiol 14:601–622
Imanaka T, Tsunekawa H, Aiba S (1980) Phenotypic stability of trp operon recombinant plasmids in E. coli. J Gen Microbiol 118:253–261
Inselburg J (1978) ColE1 plasmid mutants affecting growth of an E. coli recBrecC sbcB mutant. J Bacteriol 133:433–436
Jones IM, Primrose SB, Robinson A, Ellwood DC (1980) Maintenance of some ColE1-type plasmids in chemostat culture. Mol Gen Genet 180:579–584
Klingmüller W (1979) Genetic engineering for practical application. Naturwiss 66:182–189
Meacock PA, Cohen SN (1980) Partitioning of bacterial plasmids during cell division: a cis-acting locus that accomplishes stable plasmid inheritance. Cell 20:529–542
Melling J, Ellwood DL, Robinson A (1977) Survival of R factor carrying E. coli in mixed cultures in the chemostat. FEMS Lett 2:87–89
Roth M, Müller G, Noack D (1980) Loss of the multi-copy resistance plasmid pBR325 from Escherichia coli GY2354 pBR325 during continuous cultivation. In: Mitsuhashi S, Rosival L and Kremery V (eds) Antibiotic resistance. Avicenum and Springer Verlag, Prague, Berlin, Heidelberg, New York, p 143
Sakakibara Y, Tomizawa J (1976) Formation of catenated molecules by replication of colicin E1 plasmid DNA in cell extracts. J Mol Biol 108:569–582
Soberon X, Covarrubias L, Bolivar F (1980) Construction and characterization of new cloning vehicles. IV: Deletion derivatives of pBR322 and pBR325. Gene 9:287–305
Wouters JTM, Andel JG van (1979) R-plasmid persistence in E. coli grown in chemostat cultures. Antonie van Leeuwenhoek 45:317–318
Wouters JTM, Driehuis FL, Polaczek PJ, Oppenraay M-LHA van, Andel JG van (1980) Persistence of the pBR322 plasmid in Escherichia coli K12 grown in chemostat cultures. Antonie van Leeuwenhock 46:353–362
Zünd P, Lebek G (1980) Generation time-prolonging R plasmids: correlation between increase in the generation time of Escherichia coli caused by R plasmids and their molecular size. Plasmid 3:65–69
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Noack, D., Roth, M., Geuther, R. et al. Maintenance and genetic stability of vector plasmids pBR322 and pBR325 in Escherichia coli K12 strains grown in a chemostat. Molec. Gen. Genet. 184, 121–124 (1981). https://doi.org/10.1007/BF00271207
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DOI: https://doi.org/10.1007/BF00271207