• Title/Summary/Keyword: Genetic discrimination

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Discrimination of Bacillus subtilis from Other Bacillus Species Using Specific Oligonucleotide Primers for the Pyruvate Carboxylase and Shikimate Dehydrogenase Genes

  • Lee, Gawon;Heo, Sojeong;Kim, Tao;Na, Hong-Eun;Park, Junghyun;Lee, Eungyo;Lee, Jong-Hoon;Jeong, Do-Won
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1011-1016
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    • 2022
  • Bacillus subtilis is a useful bacterium in the food industry with applications as a starter strain for fermented food and as a probiotic. However, it is difficult to discriminate B. subtilis from other Bacillus species because of high phenotypic and genetic similarity. In this study, we employed five previously constructed multilocus sequence typing (MLST) methods for the discrimination of B. subtilis from other Bacillus species and all five MLST assays clearly distinguished B. subtilis. Additionally, the 17 housekeeping genes used in the five MLST assays also clearly distinguished B. subtilis. The pyruvate carboxylase (pyrA) and shikimate dehydrogenase (aroE) genes were selected for the discrimination of B. subtilis because of their high number of polymorphic sites and the fact that they displayed the lowest homology among the 17 housekeeping genes. Specific primer sets for the pyrA and aroE genes were designed and PCR products were specifically amplified from B. subtilis, demonstrating the high specificity of the two housekeeping genes for B. subtilis. This species-specific PCR method provides a quick, simple, powerful, and reliable alternative to conventional methods in the detection and identification of B. subtilis.

Survey on Revision and Complements for the Current Curriculum of Herbology (한의과대학 본초학 교육과정의 개정 및 보완을 위한 설문조사 연구)

  • Kim, Hong-Jun;Choi, Go-Ya;Kim, Chul;Lee, Guem-San;Kim, Jung-Hun;Lee, Seung-Ho;Hwang, Sung-Yeoun;Ju, Young-Sung
    • The Journal of Korean Medicine
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    • v.30 no.4
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    • pp.118-128
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    • 2009
  • Objects: This study was conducted to investigate the current educational environment of herbology and to develop a future-oriented curriculum for oriental medicine. The questionnaire used in this research was drawn up based on the current curriculum referring to the current curriculum of herbology and pharmacognosy. Methods: The survey was carried out presenting the questionnaires to a total 12,754 of the students and doctors of oriental medicine through e-mailing five times; of these, 2,074 replied. Results: 1. Among the respondents, about 97% agreed that it was necessary to revise and complement the current curriculum of herbology. 2. The respondents felt that the assigned lecture time of subject was "sufficient" (19%), "insufficient" (39%) and "average" (39%), respectively, and the level of lecture was "insufficient" (37%) or "average" (43%) respectively. According to priority, it showed that the contents which needed complement in lecture were discrimination of medicinal herbs (24%), practical use of action and indications (23%), and correlation with modern disease (21%). In theoretical lectures, 69% of the respondents agreed on the introduction of natural scientific methods 3. In practice, 51% of the respondents replied that the lecture time for practice was insufficient. The contents which needed to be complemented in practice were as follows: audio-visual materials for discrimination of medicinal herbs (22%), concrete exercise for the processing of medicinal herbs (21%), and attempts for the objective discrimination of medicinal herbs using instruments (microscope, analytical instrument, residual pesticide, heavy metal, genetic analysis) (16%). 70% replied that the discrimination of medicinal herbs of high price and rarity was "none or insufficient". 4. 56% replied that it was necessary to introduce and practice physicochemical analysis, and they showed higher requests according to the increase of their educational level. However, 86% replied that they had never experienced concrete attempts for objective discrimination of medicinal herbs, which seemed to indicate that, excepting some schools, practice exercise was rarely performed. Conclusions: According to results, it seems that an urgent review on the current course of herbology and a workshop on the process of experimental practice for professors is needed.

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Assessment of Sensitivity of Photo-Chromosomal Assay in the Prediction of Photo-carcinogenicity (광염색체이상시험의 광발암성 예측능력에 대한 평가)

  • Hong Mi-Young;Kim Ji-Young;Lee Young Mi;Lee Michael
    • Toxicological Research
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    • v.21 no.2
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    • pp.99-105
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    • 2005
  • Photo-mutagenic compounds have been known to alter skin cancer rates by acting as initiators or by affecting subsequent steps in carcinogenesis. The objectives of this study are to investigate the utility of photo-chromosomal aberration (photo-CA) assay for detecting photo-clastogens, and to evaluate its ability to predict rodent photocarcinogenicity. Photo-CA assay was performed with five test substances that demonstrated positive results in photo-carcinogenicity tests: 8-Methoxypsoralen (photoactive substance that forms DNA adducts in the presence of ultraviolet A irradiation), chlorpromazine (an aliphatic phenothiazine an alpha-adrenergic blocking agent), lomefloxacin (an antibiotic in a class of drugs called fluoroquinolones), anthracene (a tricyclic aromatic hydrocarbon a basic substance for production of anthraquinone, dyes, pigments, insecticides, wood preservatives and coating materials) and Retinoic acid (a retinoid compound closely related to vitamin A). For the best discrimination between the test substance-mediated genotoxicity and the undesirable genotoxicity caused by direct DNA absorption, a UV dose-response of the cells in the absence of the test substances was firstly analyzed. All 5 test substances showed a positive outcome in photo-CA assay, indicating that the photo-CA test is very sensitive to the photo-genotoxic effect of UV irradiation. With this limited data-set, an investigation into the predictive value of this photo-CA test for determining the photo-carcinogenicity showed that photo-CA assay has the high ability of a test to predict carcinogenicity. Therefore, the photo-CA test using mammalian cells seems to be a sensitive method to evaluate the photo-carcinogenic potential of new compounds.

Forensic Characterization of Four New Bovine Tri-nucleotide Microsatellite Markers in Korean Cattle (Hanwoo)

  • Sim, Yong Teak;Na, Jong Gil;Lee, Chul-Sang
    • Journal of Animal Science and Technology
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    • v.55 no.2
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    • pp.87-93
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    • 2013
  • We identified four new bovine tri-nucleotide microsatellite loci and analyzed their sequence structures and genetic parameters in 105 randomly selected Korean cattle (Hanwoo). Allele numbers of the loci B17S0808, B15S6253, B8S7996, and B17S4998 were 10, 11, 12, and 29, respectively. These alleles contained a simple or compound repeat sequences with some variations. Allele distributions of all these loci were in Hardy-Weinberg equilibrium (P > 0.05). Observed heterozygosity and expected heterozygosity ranged from 0.54 (B15S6253) to 0.92 (B17S4998) and from 0.599 (B15S6253) to 0.968 (B17S4998), respectively, and two measures of heterozygosity at each locus were highly correlated. Polymorphism information content (PIC) for these 4 loci ranged from 0.551 (B15S6253) to 0.932 (B17S4998), which means that all these loci are highly informative (PIC > 0.5). Other genetic parameters, power of discrimination (PD) and probability of exclusion (PE) ranged from 0.783 (B15S6253) to 0.984 (B17S4998) and from 0.210 (B15S6253) to 0.782 (B17S4998), respectively. Their combined PD and PE values were 0.9999968 and 0.98005176, respectively. Capillary electrophoresis revealed that average peak height ratio for a stutter was 13.89% at B17S0808, 26.67% at B15S6253, 9.09% at B8S7996, and 43.75% at B17S4998. Although the degree of genetic variability of the locus B15S6253 was relatively low among these four microsatellite markers, their favorable parameters and low peak height ratios for stutters indicate that these four new tri-nucleotide microsatellite loci could be useful multiplex PCR markers for the forensic and population genetic studies in cattle including Korean native breed.

Discrimination of Korean Ginseng Cultivars by Sequence Tagged Sites (STS) Markers (STS 마커를 이용한 고려인삼 품종 및 육성계통 판별)

  • Jo, Ick Hyun;Shin, Mi Ran;Kim, Young Chang;Lee, Seung Ho;Kim, Jang Uk;Moon, Ji Young;Noh, Bong Soo;Kang, Sung Taek;Lee, Dong Jin;Hyun, Dong Yun;Kim, Dong Hwi;Kim, Kee Hong;Bang, Kyong Hwan
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.5
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    • pp.353-360
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    • 2013
  • Korean ginseng (P. ginseng C. A. Meyer) is one of the most important medicinal plant in the world. Understanding genetic variability among the assortment of Korean ginseng is important for breeding. The aim of this study was to molecularly characterize Korean ginseng cultivar and breeding lines through the use of eight previously reported STS markers (MFGp183, MFGp130, MFGp110, UFGp74, UFGp163, MFGp108, MFGp81 and UFGp156). All STS markers produced interpretable electropherograms from 31 accessions consisting of 11 Korean ginseng cultivars and 20 breeding lines. When eight STS markers were combined, we identified to total 19 genetic patterns; in particular, nine cultivars (Chunpoong, Yunpoong, Gopoong, Gumpoong, Sunpoong, Sunone, Cheongseon, Sunhyang, Cheonryang) and 5 breeding lines (G08012, G04079, G04075, G08036, G04110) in ginseng samples can be discriminated from the others. Together with other available markers, these STS markers will contribute to the management of ginseng genetic resources and the protection of breeders' rights.

Genetic Variations of D16S539 Locus in the Korean Population (한국인에서 D16S539 유전좌의 유전적 다형성)

  • Shin, Kyoung-Jin;Yang, Yun-Seok;Choi, Jong-Hoon;Yang, Woo-Ick;Cho, Sang-Ho;Kim, Chong-Youl
    • Journal of Oral Medicine and Pain
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    • v.25 no.4
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    • pp.395-402
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    • 2000
  • The D165539 locus was investigated to collect population genetic data in the Korean population. The selected subject was unrelated 293 Korean people. DNA was extracted from the samples and PCR was performed with fluorescent primer. The amplified fragment was analysed by automated DNA sequencer and it's application software. Among the Korean population, 7 allele and 18 geneotype were observed and allele No. 9 is mostly frequent(0.2679) and then allele No. 11(0.2679), allele No. 9(0.2645). The observed heterozygosity and the expected heterozygosity is 0.7466, 0.7829 each. The polymorphism information content(PIC) is 0.7466. The power of discrimination(PD) and the mean exclusion chance(MEC) are calculated to be 0.9190 and 0.5775.

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Comparative Molecular Analysis of Freshwater Centric Diatoms with Particular Emphasis on the Nuclear Ribosomal DNA of Stephanodiscus (Bacillariophyceae)

  • Ki, Jang-Seu
    • ALGAE
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    • v.24 no.3
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    • pp.129-138
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    • 2009
  • DNA-based discrimination of species is a powerful way for morphologically otherwise similar species, like centric diatoms. Here, the author sequenced long-range nuclear ribosomal DNAs, spanning from the 18S to the D5 region of the 28S rDNA, of Stephanodiscus, particularly including a Korean isolate. By comparisons, high DNA similarities were detected from the rDNAs of nine Stephanodiscus (>99.4% in 18S rDNA, >98.0% in 28S rDNA). Their genetic distances, however, were significantly different (Kruskal-Wallis test, p < 0.01) compared to two related genera, namely Cyclotella and Discostella. In addition, genetic distances of 18S rDNAs were significantly different (Student’s t-test, p = 0.000) against those of the 28S rDNAs according to individual genera (Cyclotella, Discostella, and Stephanodiscus). Phylogenetic analyses showed that Stephanodiscus and Discostella showed a sister taxon relationship, and their clade was separated from a cluster of Cyclotella (1.00 PP, 100% BP). This suggests that Stephanodiscus has highly conserved sequences of both 18S and 28S rDNA; however, Stephanodiscus is well-separated from other freshwater centric diatoms, such as Cyclotella and Discostella, at the generic level.

Validation of Photo-comet Assay as a Model for the Prediction of Photocarcinogenicity

  • Kim, Ji-Young;Koh, Woo-Suk;Lee, Mi-Chael
    • Toxicological Research
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    • v.22 no.4
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    • pp.423-429
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    • 2006
  • Recent reports on the photocarcinogenicity and photogerotoxicity of many compounds led to an increasing awareness for the need of a standard approach to test for photogenotoxicity. The comet assay has been recently validated as a sensitive and specific test system for the quantification of DNA damage. Thus, the objectives of this study are to investigate the utility of photo-comet assay for detecting photo-mutagens, and to evaluate its ability to predict rodent photo-carcinogenicity. Photo-comet assays were performed using L5178Y $Tk^{+/-}$ mouse lymphoma cells on five test substances (8-methoxypsoralen, chlorpromazine, lomefloxacin, anthracene and retinoic acid) that demonstrated positive results in photocarcinogenicity tests. For the best discrimination between the test substance-mediated DNA damage and the undesirable DNA damage caused by direct UV absorption, a UV dose-response of the cells in the absence of the test substances was firstly fnalized. Out of 5 test substances, positive comet results were obtained for chlorpromazine, lomefloxacin, anthracene and retinoic acid while 8-methoxypsoralen found negative. An investigation into the predictive value of this photo-comet assay for determining the photocarcinogenicity showed that photo-comet assay has relatively high sensitivity. Therefore, the photo-comet assay with mammalian cells seems to be a good and sensitive predictor of the photocarcinogenic potential of new substances.

Discrimination of Panax ginseng Roots Cultivated in Different Areas in Korea Using HPLC-ELSD and Principal Component Analysis

  • Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Min-Kyung;Lee, Jae-Woong;Lee, Youn-Hyung;Yang, Deok-Chun;Baek, Nam-In
    • Journal of Ginseng Research
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    • v.35 no.1
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    • pp.31-38
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    • 2011
  • In order to distinguish the cultivation area of Panax ginseng, principal component analysis (PCA) using quantitative and qualitative data acquired from HPLC was carried out. A new HPLC method coupled with evaporative light scattering detection (HPLC-ELSD) was developed for the simultaneous quantification of ten major ginsenosides, namely $Rh_1$, $Rg_2$, $Rg_3$, $Rg_1$, Rf, Re, Rd, $Rb_2$, Rc, and $Rb_1$ in the root of P. ginseng C. A. Meyer. Simultaneous separations of these ten ginsenosides were achieved on a carbohydrate analytical column. The mobile phase consisted of acetonitrile-water-isopropanol, and acetonitrile-water-isopropanol using a gradient elution. Distinct differences in qualitative and quantitative characteristics for ginsenosides were found between the ginseng roots produced in two different Korean cultivation areas, Ganghwa and Punggi. The ginsenoside profiles obtained via HPLC analysis were subjected to PCA. PCA score plots using two principal components (PCs) showed good separation for the ginseng roots cultivated in Ganghwa and Punggi. PC1 influenced the separation, capturing 43.6% of the variance, while PC2 affected differentiation, explaining 18.0% of the variance. The highest contribution components were ginsenoside $Rg_3$ for PC1 and ginsenoside Rf for PC2. Particularly, the PCA score plot for the small ginseng roots of six-year old, each of which was light than 147 g fresh weight, showed more distinct discrimination. PC1 influenced the separation between different sample sets, capturing 51.8% of the variance, while PC2 affected differentiation, also explaining 28.0% of the variance. The highest contribution component was ginsenoside Rf for PC1 and ginsenoside $Rg_2$ for PC2. In conclusion, the HPLC-ELSD method using a carbohydrate column allowed for the simultaneous quantification of ten major ginsenosides, and PCA analysis of the ginsenoside peaks shown on the HPLC chromatogram would be a very acceptable strategy for discrimination of the cultivation area of ginseng roots.

DNA Fingerprinting Analysis of Natural Monument Gingko Trees Using Microsatellite Markers (Microsatellite 마커를 이용한 은행나무 천연기념물의 DNA 지문 분석)

  • Lee, Jei-Wan;Lee, Min-Woo;Ahn, Ji-Young;Hong, Kyung-Nak;Baek, Seung-Hoon;Kim, Sang-Chul
    • Journal of Korean Society of Forest Science
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    • v.106 no.4
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    • pp.408-416
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    • 2017
  • This study describes DNA fingerprinting analysis of twenty-three natural monument individuals of Ginkgo biloba using eight microsatellite markers. The average number of observed alleles was 6.875, and the expected heterozygosity and the observed heterozygosity were 0.711 and 0.710, respectively. This results were similar to those of the previous studies on Ginkgo trees analyzed by same markers in China and Japan. PIC value and PD were calculated at 0.677 and 0.9999 respectively, indicating a high individual identification efficiency. In fact, all of the natural monument ginkgo trees and additionally analyzed thirteen general ginkgo tress were identified by genotype comparison. PI and PD calculated in three markers (Ging06, Gb60, Gb61) with the highest PIC values calculated in natural monument ginkgo trees were $8.045{\times}10^{-5}$ and 99.99%, respectively. Thus, these three markers could be preferentially used in DNA fingerprinting for identifying ginkgo tree individuals. The results in this study will be useful for management of natural monument ginkgo trees, proliferation of their progeny and genetic identification of individuals selected in breeding process.