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Nucleic Acids Research, 1979, Vol. 6, No. 6 2109-2123
© 1979

Articles

Isolation and organization of calf ribosomal DNA

Michèle Meunier-Rotival, Jordi Cortadas*, Gabriel Macaya** and Giorgio Bernardi

Laboratoire de Génétique Moléulaire, Institut de Recherche en Biologie Moléculaire F-75221 Paris, France

Received February 14, 1979.

Ribosomal DNA (rDNA) from calf was isolated by three density gradient centrifugations. The first centrifugation in Cs2SO4/BAMD was used to obtain partially resolved dG+dC-rich fractions from total DNA. The second and third centrifugations. in Cs2SO4/Ag+, led to the isolation of an rDNA fraction characterized by a symmetrical band in CsCl, p=1.724 g/cm3. This new procedure appears to be generally suitable for the isolation of rDNA and other dG+dC-rich repeated genes.

The organization of isolated calf rDNA has been studied by restriction enzyme digestion and by hybridization with cloned rDNA from Xenopus laevis. The repeat unit of calf rDNA has a molecular weight of 21x106 and it split by EcoR1 into two fragments, 16x106 and 5.0x106, and by BamHI into seven fragments. EcoRI and BamHI sites have been mapped. Most of the 18S and 28S RNA genes and the transcribed spacer are contained in the small EcoRI fragment, while the non-transcribed spacer is localized on the large EcoRI fragment. This spacershowed length heterogeneity within a single individual; such heterogeneity is limited to two regions of the spacer.

*Present address: Departamento de Quimica Macromolecular del C.S.I.C., Escuela T.S. de Ingenieros Industriales, Diagonal 999, Barcelona-28, Spain.

**Present address: Centro de Investigación en Biologia Celular y Molecular, Universidad de Costa Rica. San José, Costa Rica.


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