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최해운,송하영,구달회,방재욱,허윤강 한국유전학회 2007 Genes & Genomics Vol.29 No.4
In order to identify and discriminate a medicinal herb, A. acutiloba, from closely related species, species-specific repetitive DNAs have been cloned and characterized by Southern blotting and FISH (fluorescence in situ hybridization). From restriction enzyme analysis, tandemly repeated sequences specific for A. acutiloba (AaTRS) were obtained from an EcoRI digestion. The EcoRI monomers were 154-159 bp long, 60-66% of A+T residues and did not show any similarity to the known plant repetitive DNAs. Its repeats are organized in head-to-tail orientation. Southern blot analysis of genomic DNA digested with the same enzyme showed the existence of a ladder pattern only in A. acutiloba, indicating that the repetitive DNA is arrayed in tandem. In FISH experiment, the AaTRS was localized on the telomeric regions of somatic metaphase chromosomes in A. acutiloba. Although seven monomers revealed high sequence variation as the typical features of eukaryotic repetitive DNA, SCAR (sequence characterized amplified region) markers generated from their sequence information could discriminate A. acutiloba from two other species.