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• W1990444549 abstract "Abstract We recently reported that the six-megadalton plasmid pAMα1 (one of two plasmids in Streptococcus faecalis strain DS-5C1) undergoes a gene amplification as a result of extended growth (many generations) of cells in the presence of a subinhibitory concentration of tetracycline. In this communication we present a detailed analysis of this phenomenon using the newly constructed S. faecalis strain DT-11, a strain devoid of plasmid DNA other than pAMα1. It was found that during growth of strain DT-11 in the presence of tetracycline a gradual increase in plasmid size was observed, and this could be correlated with an increase in the level of tetracycline resistance. On the basis of analyses using sucrose density-gradient sedimentation, electron microscopy and the restriction endonuclease EcoRI, it was clearly established that the enlarged DNA contained repeated units of a 2.65 megadalton segment containing the tetracycline resistance determinant. This value (2.65 megadaltons) also corresponds to the amount of DNA deleted from the plasmid in easily obtainable tetracycline sensitive variants. EcoRI digestion of the amplified DNA structures generated three fragments designated α1,α2 and α3 with molecular weights of 4.20 × 106, 1.75 × 106 and 0.90 × 106, respectively. Unamplified pAMα1 was cleaved in only two places by the enzyme, while the plasmid of tetracycline sensitive variants (i.e. DNA missing the 2.65 megadalton sequence on which the tetracycline resistance determinant resides) is totally insensitive. In the case of amplified molecules the relative amount of the α2 fragment with respect to the α1 fragment is much higher than for normal pAMα1. The data indicate that the α2 fragment contains the tetracycline resistance determinant; and in the amplified state the additional copies are arranged in tandem connected together by the α3 fragment(s). The data support models which we proposed previously with regard to the mechanism of amplification. It was also observed that in the amplified state the total amount of closed circular plasmid DNA relative to chromosomal DNA was found to be similar to that for the case of unamplified DNA, indicating that when the molecules are larger there are fewer of them." @default.
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• W1990444549 date "1976-04-01" @default.
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• W1990444549 title "Plasmid-determined tetracycline resistance in Streptococcus faecalis: Tandemly repeated resistance determinants in amplified forms of pAMα1 DNA" @default.
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• W1990444549 doi "https://doi.org/10.1016/0022-2836(76)90336-3" @default.
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