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TN5 insertions in ribosomal protein genes in Escherichia coli Hui, Ivy Yuk Chee


The genetic organization and interrelationship between the two ribosomal protein transcription units (the L11 and the L10 operons) from near 89 min on the Escherichia coli chromosome were studied using insertional mutations generated by the kanamycin resistant transposable element Tn5. The polar effects of Tn5 insertion on the expression of these ribosomal genes were examined by: (1) The level of β-galactosidase generated from L10-lacZ and β-lacZ fusion proteins; (2) Direct SDS-polyacrylamide gel electrophoresis of the proteins specified -by the plasmid ribosomal genes in UV-inactivated maxicells; (3) Urea-polyacrylamide gel electrophoresis of plasmid and chromosome specified L12 proteins in an L12 electrophoretic mutant (Strain JF1039). The above experiments localized the major promoter for the L10 operon between nucleotide 1280 and 1360. These investigations also confirmed that the translational regulatory proteins for the L11 and L10 operons are L1 and L10 respectively.

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