• Title/Summary/Keyword: DNA Marker

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Development of Molecular Markers for the authentication of Zanthoxyli Pericarpium by the analysis of rDNA-ITS DNA barcode regions (rDNA-ITS DNA 바코드 부위 분석을 통한 산초(山椒) 기원종 감별용 유전자 마커 개발)

  • Kim, Wook Jin;Ji, Yunui;Lee, Young Mi;Kang, Young Min;Choi, Goya;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.30 no.3
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    • pp.41-47
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    • 2015
  • Objectives : Due to the morphological similarity of the pericarp and description of multi-species in National Pharmacopoeia of Korea and China, the Zanthoxylum Pericarpium is difficult to authenticate adulterant in species levels. Therefore, we introduced the sequence analysis of DNA barcode and identification of single nucleotide polymorphism(SNP) to establish a reliable tool for the distinction of Zanthoxylum Pericarpium from its adulterants. Methods : To analyze DNA barcode region, genomic DNA was extracted from twenty-four specimens of authentic Zanthoxylum species and inauthentic adulterant and the individual internal transcribed spacer regions (rDNA-ITS and ITS2) of nuclear ribosomal RNA gene were amplified using ITS1, ITS2-S2F, and ITS4 primer. For identification of species-specific sequences, a comparative analysis was performed using entire DNA barcode sequences. Results : In comparison of four Zanthoxylum ITS2 sequences, we identified 16, 4, 6, and 4 distinct species-specific nucleotides enough to distinguish Z. schinifolium, Z. bungeanum, Z. piperitum, and Z. simulans, respectively. The sequence differences were available genetic marker to discriminate four species. Futhermore, phylogenetic relationship revealed a clear classification between different Zanthoxylum species showing 4 different clusters. These results indicated that comparative analysis of ITS2 DNA barcode was an useful genetic marker to authenticate Zanthoxylum Pericarpium in species levels. Conclusions : The marker nucleotides, enough to distinguish Z. schinifolium, Z. piperitum, Z. bungeanum, and Z. simulans, were obtained at 30 SNP marker nucleotides from ITS2 sequences. These differences could be used to authenticate official Zanthoxylum Pericarpium from its adulterants as well as discriminating each four species.

Development of a Female-associated SCAR Marker in Schisandra nigra Max. (Schisandra nigra Max.에서 암그루에 연관된 SCAR 마커의 개발)

  • Han, Hyo Shim;Jung, Jae Sung
    • Journal of Life Science
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    • v.31 no.6
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    • pp.537-542
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    • 2021
  • Schisandra nigra Max., a dioecious plant native to Jeju Island in Korea, is cultivated on a small scale for fruit production. As fruit-producing female individuals are generally considered to be more valuable than male, early identification of plant sex at the seedling stage is important. In this study, a sequence-characterized amplified region (SCAR) marker associated with a female-specific region in the genome of S. nigra was investigated. Of 120 randomly amplified polymorphic DNA (RAPD) primers, one primer (OPB-03) consistently amplified a 749 bp band in female plants. The female-specific PCR product was isolated and cloned, and the nucleotide sequences were then determined. Southern hybridization performed using the female-specific fragment as a probe produced positive signals only in genomic DNA from the female plants. This result revealed that the 749 bp segment of DNA was present in the genome of female plants but absent in the genome of male plants. A SCAR primer pair was designed based on the RAPD marker to amplify a 436 bp fragment in the genomic DNA of female plants. This primer pair amplified the expected size of DNA fragment in female plants and four monoecious individuals collected from a natural population. The SCAR marker identified in this study can be used to distinguish female-flowering individuals at the seedling stage.

Discrimination of Korean Native Chicken Populations Using SNPs from mtDNA and MHC Polymorphisms

  • Hoque, M.R.;Lee, S.H.;Jung, K.C.;Kang, B.S.;Park, M.N.;Lim, H.K.;Choi, K.D.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.12
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    • pp.1637-1643
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    • 2011
  • Korean native chickens are a very valuable chicken population in Korea and their prices are higher than that of commercial broilers. In order to discriminate two commercial Korean native chicken populations (CCP1 and CCP2), single nucleotide polymorphisms (SNPs) from mitochondrial (mt) DNA D-loop sequences and LEI0258 marker polymorphisms in the major histocompatibility complex (MHC) region were investigated. A total of 718 birds from nine populations were sampled and 432 mtDNA sequences were obtained. Of these, two commercial Korean native chicken populations (363 birds) were used for investigation of their genetic relationship and breed differentiation. The sequence data classified the chickens into 20 clades, with the largest number of birds represented in clade 1. Analysis of the clade distribution indicated the genetic diversity and relation among the populations. Based on the mtDNA sequence analysis, three selected SNPs from mtDNA polymorphisms were used for the breed identification. The combination of identification probability (Pi) between CCP1 and CCP2 using SNPs from mtDNA and LEI0258 marker polymorphisms was 86.9% and 86.1%, respectively, indicating the utility of these markers for breed identification. The results will be applicable in designing breeding and conservation strategies for the Korean native chicken populations and also used for the development of breed identification markers.

Molecular Authentication of Pinelliae Tuber from its adulterants by the analysis of DNA barcodes, matK and rbcL genes (matK와 rbcL DNA 바코드 분석을 통한 반하(半夏) 및 반하(半夏) 유사 한약재 유전자 감별)

  • Lee, Young Mi;Moon, Byeong Cheol;Ji, Yunui;Kim, Wook Jin;Kim, Ho Kyoung
    • The Korea Journal of Herbology
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    • v.28 no.6
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    • pp.53-58
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    • 2013
  • Objectives : Pinelliae Tuber has been used as a typical unauthentic herbal medicines. Due to the morphological similarity between Pinelliae Tuber and adulterants, the correct authentication is very difficult. Therefore, we introduced DNA barcode to establish a powerful tool for the authentication of Pinelliae Tuner from adulterants. Methods : To obtain DNA barcode regions, genomic DNA was extracted from nineteen specimens of Pinellia ternata, Pinellia pedatisecta, Pinellia tripartita, and Typhonium flagelliforme, and matK and rbcL genes were amplified. For identification of species specific sequences and analysis phylogenetic relationship, a comparative analysis were performed by the ClastalW and UPGMA based on entire sequences of matK and rbcL genes, respectively. Results : In comparison of two DNA barcode sequences, we elucidated the phylogenetic relationship showing distinct four groups depending on species and identified 40 and 20 species specific nucleotides enough to distinguish each species from matK and rbcL gene, respectively. The sequence differences at the corresponding positions were avaliable genetic marker nulceotides to discriminate the correct species among analyzed four species. These results indicated that phylogentic and comparative analysis of matK and rbcL genes are useful genetic markers to authenticate Pinelliae Tubers. Conclusions : The marker nucleotides enough to distinguish P. ternata, P. tripatrita, P. peditisecta, and T. flagelliform, were observed at 40 positions in matK gene and 20 positions in rbcL gene sequence, respectively. These differences can be used to authenticate Pinelliae Tuber from adulterants as well as discriminate each four species.

Identification of DNA Markers Related to Resistance to Herbicide Containing Mesotrione in Tongil Type Rice (통일형 벼에서 메소트리온계 제초제 저항성 연관 DNA marker 탐색)

  • Lee, Ji-Yoon;Cho, Jun-Hyeon;Lee, Jong-Hee;Cho, Su-Min;Kwon, Young-Ho;Park, Dong-Soo;Song, You-Chun;Ko, Jong-Min
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.387-395
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    • 2018
  • This study was conducted to identify DNA markers related to resistance to herbicide containing mesotrione in Tongil type rice. Two Tongil type elite lines; Milyang154 and Suweon382, showed resistance to mesotrione, whereas the others were susceptible at 20 days after mesotrione application, and severe growth inhibition was observed in the remaining 13 lines. As a result of analysis of mesotrione resistance using 190 $F_2$ populations derived from a cross of Hanareum2 (susceptible) and Milyang154 (resistant), the mesotrione resistance locus was shown to be a single dominant gene with a 3:1 segregation ratio ($X^2=1.19$, P=0.31). To identify a DNA marker closely linked to the mesotrione resistance gene, bulked segregant analysis (BSA) was adopted. The DNA marker RM3501 was identified on chromosome 2 with a recombinant value of 0.53 to the mesotrione resistance gene. Mst1(t) was located between SSR (simple sequence repeat) markers RM3501 and RM324 with a physical map distance of 10.2 Mb-11.4 Mb on chromosome 2. The band pattern of agarose gel electrophoresis of the SSR marker RM3501 showed the same segregation pattern with respect to mesotrione treatment in 20 Tongil type varieties and a $BC_2F_2$ segregation population derived from a cross between Unkwang (resistant) and Hanareum2 (susceptible). Thus, the RM3501 DNA marker could be used in breeding programs for Marker Assisted Selection in mesotrione resistant rice breeding.

Analysis of Genetic Characteristics of Korean Native Chicken Using DNA Marker (DNA Marker를 이용한 한국 재래닭의 유전특성 분석)

  • 이학교;이성진;황규춘;정일정;박용호;손시환;신영수;오봉국;한재용
    • Korean Journal of Poultry Science
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    • v.23 no.4
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    • pp.177-183
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    • 1996
  • This study was conducted to analyze genetic characteristics of Korean Native Chicken three lines classified on the basis of the feather color and appearance (Red, Yellow, and Black) using DNA fingerprinting method. To estimate the genetic relatedness among breeds and similarities within breeds, we collected blood samples from Korean Native Chicken (KNC), Rhode Island Red (RIR), White Leghorn (WL), and Cornish(CN) and obtained genomic DNA from the blood of 10 individuals randomly selected within the breeds and lines. The genomic DNA samples were digested with restriction enzymes (Hinf J, Hae Ill) and hybridized with various probes (Jeffreys' probes 33.15, 33.6 and M13) after Southern transfer. Genetic similarities within breeds were characterized by band sharing (BS) value, estimated by the DFP band pattern between the pair of lanes. BS values within WL, RIR, and KNC were 0.82, 0.70 and 0.56, respectively. Relative genetic diversity (BS value) of KNC was higher than those two breeds (WL, RIR). Estimation of genetic similarity between KNC lines and control breed (RIR) was 0.32, whereas similarity within KNC lines (6 groups) was 0.50. In this analysis, KNC was showed to have a highly genetic diver-sity at the DNA level, and to be closer in genetic distance to RIR (0.67) than any other breeds.

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Nonlinear matching measure for the analysis of on-off type microarray image (온-오프 형태의 DNA 마이크로어레이 영상 분석을 위한 비선형 정합도)

  • Ryu Mun ho;Kim Jong dae
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.30 no.3C
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    • pp.112-118
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    • 2005
  • In this paper, we propose a new nonlinear matching measure for automatic analysis of the on-off type DNA microarray images in which the hybridized spots are detected by the template matching method. The proposed measure is obtained by binary-thresholding over the whole template region and taking the number of white pixels inside the spotted area. This measure is compared with the normalized covariance in terms of the classification ability of the successfulness of the locating markers. The proposed measure is evaluated for the scanned images of HPV DNA microarrays where the marker locating is a critical issue because of the small number of spots. The targeting spots of HPV DNA chips are designed for genotyping 22 types of the human papilloma virus(HPV). The proposed measure is proven to give more discriminative response reducing the miss cases of the successful marker locating.