Bidirectional polarity of IS2 elements and the polar effect of an IS5 insertion in the argECBH gene cluster of Escherichia coli [proceedings].
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Biomedical subjects
Publications and source records attributed to D Charlier.
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The isolation of a new type of gamma transducing phage carrying the bipolar argECBH operon of E. coli K12 is described. The argECBH segment is inserted in the phage in a direction which is opposite from that of previously isolated argECBH-carrying phages. A colE1 argECBH plasmid has been constructed. DNA fragments resulting from digestion of these genetic elements with Eco RI and Hind III restriction enzymes have been characterized by agarose gel electrophoresis and electron microscopy, including hetero-duplex analysis. Two fragments are of special significance for studies on the control of arginine synthesis, one of length 9.8 kilobases carrying the whole argECBH region, the other of length 2 kilobases carrying most or all of the control region between argE and argC.
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The accumulation or ornithine, citrulline, and possibly acetylornithine by Escherichia coli K-12 arginineless mutants provided with acetylarginine as source of arginine causes severe growth inhibition. This occurs under conditions where comparable derivatives of E. coli W (Bollon and Vogel, 1973) show little or no growth inhibition. The same conditions, which have been reported to cause noncorrelative synthesis of acetylornithinase and argininosuccinase in E. coli W (Bollon and Vogel, 1973), do not alter the correlative pattern of enzyme synthesis observed in E. coli K-12. Moreover, previously reported effects of ornithine and citrulline on repression of the arginine regulon in E. coli W are not observed in the K-12 strains examined. The bearing of these observations on possible differences between the mechanism of enzyme repression operating in the two types of strains cannot yet be fully evaluated; it is, however, clear that considerable care should be exercised before extrapolating the results obtained with one type of strain to the other one. The particularly strong inhibition of acetylarginine utilization exerted by ornithine in E. coli K-12 allows the forward selection of several classes of arginine auxotrophs from strains deficient in carbamoylphosphate biosynthesis and thus capable of ornithine accumulation. Possible applications of this technique to the genetic analysis of the bipolar argECBH operon are discussed.
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