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KMID : 0368419990420020133
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Journal of Plant Biology
1999 Volume.42 No. 2 p.133 ~ p.138
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Cloning and Structural Analyses of Partial Nuclear rDNA from Bupleurum euphorbioides(Apiaceae)
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Kim Chang-Kyun
Choi Hong-Keun
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Abstract
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For the cloning of nuclear ribosomal DNA (rDNA) from Bupleurum euphorbioides (Apiaceae), ten clones were screened by DNA-DNA hybridization method. Among them, two clones were strongly hybridized with a heterologous probe of rice rDNA and with an autologous probe of an internally-transcribed region of B. euphorbioides amplified by PCR. We sequenced both ends of the two genomic clones aligned with a known sequence of rDNA. ITS2 sequences of the two clones showed 98% and 83% homology with the ITS2 sequence of B. euphorbioides. Our clones showed 1bp and 3bp nucleotide substitutions in the 25S and intergenic spacer regions, respectively, and the ITS1 and 18S regions were both missing. Restriction enzyme sites and the orientation of both clones were analyzed for physical mapping purposes. Apart from the length difference between the two clones, we found restriction site variations in the 25S and intergenic spacer regions.
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