• 대한본초학회지
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• 제37권5호
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• pp.9-16
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• 2022

Objectives : The purpose of this study is to report that applying the genetic discrimination method to Pinelliae Tuber is suitable as a countermeasure for the limitations of morphological identification announced publicly in the Ministry of Food and Drug Safety(MFDS). Methods : Randomly selected fifty samples in Pinelliae Tuber imported from China were used for morphological and genetic identification. The morphological identification was applied method announced publicly by the MFDS. The traits of morphological identification were classified as Pinellia ternata, P. tripartita, Pinellia pedatisecta, and Typhonium flagelliforme, according to the formation of tuberous root and tuber morphology. The genetic identifications were conducted by Sequence Characterized Amplified Region(SCAR) marker and DNA barcoding analysis for cross-validation, respectively. SCAR marker was verified according to the presence or absence of amplicon through PCR amplification using species-specific primers. DNA barcoding analysis used sequence information of the matK region. Results : As a result of the morphological identification, 27 out of 50 samples were identified as original species 'P. ternata' of genuine 'Pinelliae Tuber', and 23 were identified as adulterant species 'P. pedatisecta'. Unlike this, the genetic identification was identified as the original species 'P. ternata' in all 50 samples in the SCAR marker and matK regional sequence analysis. Conclusions : Pinelliae Tuber of morphological mutant that can not be classified by morphological identification is imported from China. The SCAR marker would be used as accurate and efficient assays for species identification of the morphological mutant.

• Molecules and Cells
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• 제42권2호
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• pp.104-112
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• 2019

Tracking the fate of individual cells and their progeny through lineage tracing has been widely used to investigate various biological processes including embryonic development, homeostatic tissue turnover, and stem cell function in regeneration and disease. Conventional lineage tracing involves the marking of cells either with dyes or nucleoside analogues or genetic marking with fluorescent and/or colorimetric protein reporters. Both are imaging-based approaches that have played a crucial role in the field of developmental biology as well as adult stem cell biology. However, imaging-based lineage tracing approaches are limited by their scalability and the lack of molecular information underlying fate transitions. Recently, computational biology approaches have been combined with diverse tracing methods to overcome these limitations and so provide high-order scalability and a wealth of molecular information. In this review, we will introduce such novel computational methods, starting from single-cell RNA sequencing-based lineage analysis to DNA barcoding or genetic scar analysis. These novel approaches are complementary to conventional imaging-based approaches and enable us to study the lineage relationships of numerous cell types during vertebrate, and in particular human, development and disease.

• 대한본초학회지
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• 제34권5호
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• pp.13-20
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• 2019

Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.