Identification and characterisation of a transducing phage containing the gene for ribosomal protein S1 of Escherichia coli

Noble, Jonathan Charles Stirling (1982) Identification and characterisation of a transducing phage containing the gene for ribosomal protein S1 of Escherichia coli. Masters thesis, Memorial University of Newfoundland.

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Abstract

The recent mapping for the gene for ribosomal protein S1 (rpsA) at 20 minutes on the E. coli chromosome (Ono, M., Michihiko, K. & Mizushima, S. (1979) Mol. Gen. Genet. 174, 11-15), has been exploited in studies aimed at the identification of a transducing phage carrying this locus. The coding capacities of a λaroA transducing phage containing a portion of this chromosomal region (R. Weisberg, personal communication), have been analysed in an UVL irradiation and infection system by a combination of techniques of immunoprecipitation and SDS and two dimensional polyacrylamide gel electrophoresis. This analysis has indicated that among the six or so bacterial proteins encoded by this phage, one comigrating with pure S1 at pI 5-6 and Mr = 70,000 reacts specifically with antiserum developed against the purified protein. This species which is identified as ribosomal protein S1 by these criteria, appears to be expressed under the direction of a bacterial promoter. -- A detailed restriction map of the genome of the λaroA transducing phage has been constructed from the results of numerous single and double enzyme digestions. This information has permitted an analysis of the relationship between the arrangement of bacterial DNA sequences in the insert contained by the transducing phage and that of the chromosomal region from which it was derived. The similarity of structure indicated by this analysis supports identification of the bacterial activity governing expression of rpsA in the phage context as the normal chromosomal promoter for this gene.

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