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Characterization of two circular plasmids from the marine diatom Cylindrotheca fusiformis: plasmids hybridize to chloroplast and nuclear DNA.
Authors:Jerry D. Jacobs   James R. Ludwig   Mark Hildebrand   Allen Kukel   Teng-Yung Feng   Robin W. Ord  Benjamin E. Volcani
Affiliation:(1) Scripps Institution of Oceanography, and Center for Molecular Genetics, University of California, 92093 San Diego, La Jolla, CA, USA;(2) Present address: Department of Zoology, University of Washington, 98195 Seattle, WA, USA;(3) Present address: Hybritech, Inc., 92196-9006 San Diego, CA, USA;(4) Present address: Institute of Botany, 11529 Academia Sinica, Taipei, Taiwan, Republic of China;(5) Present address: Ministry of Agriculture and Fisheries, Ruakura (Hamilton), New Zealand
Abstract:Summary This paper reports the discovery and initial characterization of two small plasmids, pCfl and pCf2, in the marine diatomCylindrotheca fusiformis. Extracted diatom DNA separates into two bands in CsCI-Hoechst 33258 dye gradients. Upon agarose gel electrophoresis of a sample of the upper band of the gradient we observed, in addition to high molecular weight (genomic) chloroplast and mitochondrial DNA, pairs of lower molecular weight bands. These bands contained two species of circular plasmid DNA molecules, as shown by electron microscopy. The nucleotide composition of the plasmids, and chloroplast and mitochondrial DNAs is similar, as indicated by their co-banding in the gradients. They were cloned, and their restriction maps determined, showing that pCfl is 4.27 and pCf2 4.08 kb in size. By hybridization analysis, we showed that pCfl and pCf2 share regions of similarity, but not identity. Neither plasmid hybridizes with mitochondrial DNA. Both plasmids hybridize with chloroplast DNA, and pCf2 also hybridizes with nuclear DNA.
Keywords:Diatoms  Cylindrotheca fusiformis  Plasmids  Hybridization
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