Abstract
We have constructed recombinant plasmids containing the entire Escherichia coli rrnB ribosomal RNA operon and segments thereof. Cloning of the 7.5-kb BamHI fragment, from λrif d18 which contains this operon, in plasmid vectors pBR 313 or pBR 322 is described. The 3.2-kb Eco RI Bam HI fragment containing the 3′ two-thirds of the 23 S rRNA gene, the 5S rRNA gene, and the terminator region has been cloned separately in pBR 313. As the nucleotide sequences of pBR 322 and the 7.5-kb fragment carrying the rrnB operon have been established, the entire 11.9-kb sequence of pKK 3535 is now known. This makes possible precise rearrangements and site-specific alterations of the ribosomal RNA operon; thus, pKK 3535 becomes a powerful tool for studies such as initiation and termination of transcription, processing of rRNA precursors, and investigations of the structure, function, and assembly of the ribosome itself. A detailed physical map of pKK 3535 is presented.
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