Construction of Genomic DNA Library of Korean Ginseng (Panax ginseng C. A. MEYER) and Development of Sequence-Tagged Sites

This study describes an efficient approach for developing sequence tagged sites (STS) for Panax ginseng C.A. MEYER, and their applications for line discrimination. By using the methylation filtering (MF) technique, a genomic library was constructed, in which clone inserts were derived from the hypom...

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Bibliographic Details
Published in:Biological & Pharmaceutical Bulletin 2010/09/01, Vol.33(9), pp.1579-1588
Main Authors: Bang, Kyong-Hwan, Lee, Jei-Wan, Kim, Young-Chang, Kim, Dong-Hwi, Lee, Eung-Ho, Jeung, Ji-Ung
Format: Article
Language:English
Subjects:
cleaved amplified polymorphic sequence
Gene Library
Genome, Plant - genetics
Korea
methylation filtering
Panax
Panax - genetics
Panax ginseng
Plant Extracts - genetics
Plant Extracts - isolation & purification
Plant Leaves - genetics
Seeds - genetics
Sequence Tagged Sites
sequence-tagged site
specific locus tagging
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Summary:This study describes an efficient approach for developing sequence tagged sites (STS) for Panax ginseng C.A. MEYER, and their applications for line discrimination. By using the methylation filtering (MF) technique, a genomic library was constructed, in which clone inserts were derived from the hypomethylated regions of ginseng genome. A methylation unfiltered genomic library was also constructed and the clone inserts were compared to those from the MF library in terms of sequence characteristics. Sequence analysis revealed that MF efficiently enriched the protein coding region of P. ginseng, for which the repetitive DNA appeared to be as little as 2.5 fold lower than clones in the unfiltered library, and also indicated that the P. ginseng genome may contain a large fraction of methylated repetitive DNA elements. A total of 99 and 100 highly stringent STS primer sets were designed from the filtered and unfiltered library, respectively. Amplification products were tested for latent polymorphism across six cultivars of P. ginseng and other 2 Panax species using six endonucleases recognizing four-bases. STS primer sets described here will be useful for marker-assisted selection, genome mapping and line discrimination of P. ginseng or its cultivars from other Panax species.
ISSN:0918-6158
1347-5215
1347-5215
DOI:10.1248/bpb.33.1579