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Nucleic Acids Research, 1981, Vol. 9, No. 3 503-517
© 1981

MOLECULAR BIOLOGY

Isolation, characterization and restriction endonuclease mapping of the Petunia hybrida chloroplast DNA

W.A. Bovenberg, A.J. Kool and H.J.J. Nijkamp

Department of Genetics, Biological Laboratory, Vrije Universiteit De Boelelaan 1087, 1007 MC Amsterdam, The Netherlands

Received December 9, 1980. A procedure is developed for the isolation of intact chloroplast DNA (ctDNA) from Petunia hybrida. The molecular weight, calculated from contour length measurements, is 96.0 ± 4.5 x 106 daltons. This value is in good agreement with the value of 101.2 x 106 daltons that was estimated from the electrophoretic mobilities of restriction endonuclease fragments of ctDNA. Analysis of petunia ctDNA in neutral CsCl gradients revealed the presence of only one type of DNA at a buoyant density of 1.6987 ± 0.0005 gcm-3. This corresponds with a GC-content of 39.3 ± 0.5%. A physical map of petunia ctDNA was constructed by using the restriction endonucleases Sal I, Bgl I, Hpa I and Kpn I. The map indicates that petunia ctDNA contains two copies of a sequence in an inverted orientation. The inverted repeat regions have a minimum length of 10 x 106 daltons. Hybridization data indicate that part of the inverted repeat regions contain the genes for chloroplast ribosomal RNAs.


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