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A study of the organisation of the ribosomal ribonucleic acid gene cluster of Neurospora crassa by means of restriction endonuclease analysis and cloning in bacteriophage lambda

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Summary

1. Total Neurospora crassa DNA was restricted with endonucleases and fragments carrying rRNA coding sequences were identified by hybridization with Xenopus laevis ribosomal DNA probes. 2. The repeating unit of the rRNA gene cluster was found to be 8.6 kbp, arranged in a head-to-tail fashion. 3. Digestion with Hind III yielded fragments of 3.4 kbp and 5.2 kbp and both were cloned. 4. Digestion with Eco RI yieled fragments of 2.2 kbp, 3.0 kbp and 3.4 kbp; the 3.0 kbp fragment was cloned. 5. Sequences coding for RNA (S-rRNA)1 of the smaller subribosomal particle were found (at least 90%) in the 2.2 kbp EcoRI subfragment of the 5.2 kbp Hind III fragment. 6. The coding sequences for the major RNA species (L-rRNA) of the larger subribosomal particle were located mainly (at least 95%) in the 3.4kbp Hind III fragment. 7. For comparison, a Hind III digest of total yeast DNA was cloned and recombinants containing a 6.4 kbp rDNA fragment were isolated.

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Author notes

  1. Robert A. Cox

    Present address: National Institute for Medical Research, Mill Hill, NW7 1AA, London, UK

  2. Keith Peden

    Present address: Department of Microbiology, The Johns Hopkins University, School of Medicine, 21205, Baltimore, Md., USA

Authors and Affiliations

  1. Department of Molecular Biology, The University, West Mains Road, Edinburgh, Scotland

    Robert A. Cox & Keith Peden

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  1. Robert A. Cox
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  2. Keith Peden
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Additional information

Communicated by Ch. Auerbach

Containment Facilities. The isolation of recombinant plasmids and phage were carried out under Category 2 conditions as described in the Williams report. Recombinant DNA was handled under Category 1 conditions. These procedures were approved by GMAG and the Departmental Safety Committee.

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Cox, R.A., Peden, K. A study of the organisation of the ribosomal ribonucleic acid gene cluster of Neurospora crassa by means of restriction endonuclease analysis and cloning in bacteriophage lambda. Molec. Gen. Genet. 174, 17–24 (1979). https://doi.org/10.1007/BF00433300

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  • DOI: https://doi.org/10.1007/BF00433300

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