• Title/Summary/Keyword: 염기 단편

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Isolation and Characterization of a Chitin Synthase Gene Fragments from Pleurotus sajor-caju (여름느타리의 Chitin synthase 유전자 단편분리 및 발현 특성 분석)

  • Jeong, Mi-Jeong;Park, Soo-Chul;Kim, Bum-Gi;Yoo, Young-Bok;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.354-360
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    • 1998
  • We isolated three amplified DNA fragments from P. sajor-caju by Polmerase chain reaction (PCR) using the chithin synthase specific primers. Since the sequence analysis of the these fragments showed significant homology to the other known chitin synthase gene, we regarded these cloned fragments as PsCHS1, PsCHS2, and PsCHS3 according to their size. The PsCHS3, which showed the highest sequence homology (83% identity in amino acid level with ChsI of Rhizopus oligosporus in conserved region), was selected to see expression pattern of the corresponding gene. The result of RT-PCR using internal primer of the PsCHS3 fragment revealed that PsCHS3 gene was only expressed in cap and mycelium but not in stipe. In order to see whether the PsCHS3 gene was to be induced by wounding, the comparison of the mRNA level of this gene between wounded and unwounded mature cap showed at least two times induction of this gene by wounding treatment.

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Development of molecular marker for species authentication of Dendranthema indicum (L.) Des Moul. and D. boreale (Makino) Ling ex Kitam. (감국(Dendranthema indicum (L.) Des Moul.) 및 산국(D. boreale (Makino) Ling ex Kitam.)의 종판별 분자마커 개발)

  • Byeon, Jihui
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.66-66
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    • 2018
  • 국화과(Compositae) 다년생 초본인 산국속(Dendranthema)은 국내 약 13여종이 자생하는 것으로 알려져 있으며, 이 중 감국(D. indicum (L.) Des Moul.)과 산국(D. boreale (Makino) Ling ex Kitam.), 구절초(D. zawadskii var. latilobum (Maxim.) Kitam.)가 주로 차 또는 한약재 등의 원료로 이용되고 있다. 차로 이용되는 꽃은 산국이 감국에 비해 상대적으로 작아서 구분이 가능하지만 시중에는 건조된 형태로 가공 유통되므로 육안으로 구분이 쉽지 않고, 산국 유래 제품들은 국내에서 감국 또는 국화로 혼용해서 표기되어 유통되고 있어 그 기원을 명확히 정립할 필요가 있다. 이에 본 연구는 감국과 산국의 분자유전학적 판별을 위해 DNA 바코드 후보 유전자를 활용하여 염기서열분석으로 확보된 SNP 및 InDel 정보를 바탕으로 CAPS 마커를 개발하고자 수행되었다. 감국과 산국 모두 trnL-trnF intergenic spacer 구간에서 약 1kb의 PCR 산물이 확인되었고, 이들 염기서열에서 분석한 2 SNP 및 3 InDel을 대상으로 CAPS 마커 개발을 위한 제한효소 사이트를 탐색하였다. Gap을 포함한 774bp (감국/산국=A/G) 위치의 SNP에서 BstUI(GC^GC)처리로 CAPS 마커로 전환 가능함이 확인되었고, 이에 감국과 산국의 PCR 산물에 제한효소를 처리한 결과, 제한효소 인식 사이트가 존재하는 산국에서 두 개의 DNA 단편이 확인되었다. 위 결과는 다양한 형태로 가공 유통되는 감국과 산국의 판별을 위한 마커로 활용될 수 있으며, 본 연구에 활용된 기술은 추후 건강기능식품 개발을 위한 원료표준화 확립 연구에 유용할 것으로 판단된다.

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Characteristics of β-casein Gene using the PCR Technique in Korean Native Goat (PCR 기법을 이용한 한국재래산양 β-casein 유전자의 특성)

  • Kim, Ji-Ae;Ryoo, Seung-Heui;Yu, Sung-Lan;Lee, Jun-Heon;Seo, Gil-Woong;Kim, Sun-Kyun;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
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    • v.29 no.2
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    • pp.43-52
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    • 2002
  • This study was performed to provide the basic data for preservation and improvement of genetic resources according to finding genetic construction obtained from analysis of genetic characteristics of $\beta$-casein gene in Korean Native goat and Saanen using the PCR-RFLP. This study confirmed the amplified products of 481bp fragments obtained from the amplification of $\beta$-casein loci by PCR. The $\beta$-casein AB genotype showed 481, 284 and 197bp, and $\beta$-casein BB genotype showed 284 and 197bp fragments in Korean Native goat and Saanen. The frequencies of $\beta$-casein genotype in Korean Native goat were 6.25 and 93.75% for AA and AB and the frequencies of $\beta$-casein genotype in Saanen were 57.14 and 42.86% for AA and AB types. The frequencies of $\beta$-casein A and B alleles were 0.031 and 0.969 in Korean Native goat and the frequencies of $\beta$-casein A and B alleles are 0.286 and 0.714 in Saanen, respectively. The nucleotide sequence of $\beta$-casein gene of Korean Native goat was 97.71% higher homology with 11 nucleotide sequences difference of that of goat reported in GeneBank (M90556). Therefore, this study of molecular genetic characteristics by the analysis of genetic polymorphism and sequencing for $\beta$-casein gene should be used as basic and applying data for preservation and improvement of genetic resources in Korean Native goat breeding.

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Identification of Hanwoo Using 3'-tailed Primer Associated with Single Nucleotide Polymorphism(SNP) in Melanocortin 1 Receptor(MC1R) gene (소 모색관련 MC1R 유전자의 SNP와 관련한 3'-tailed primer를 이용한 한우육의 판별)

  • Kim, T.J.;Park , S.D.;Lee , J. I.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.897-902
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    • 2004
  • To improve the methods used for the identification of Hanwoo, we performed a PCR using 3 -tailed primer associated with single nucleotide polymorphism(SNP) in Melanocortin 1 receptor(MCIR) gene. MCIR plays an important role in melanin synthesis and the SNP within MCIR was used as a target for PCRRFLP studies previously. A forward 3 -tailed primer, which matches with the template DNA of Hanwoo but not with others(blackhaired; Holstein and Black angus) at the site of 594th base sequence, and one reverse primer were designed for this study. When use this primer set, a size of 343bp was amplified by PCR only in Hanwoo, not in Holstein and Black angus. This result suggests that the PCR using our 3 -tailed primer would be very accurate, easy, reproducible and economic method to discriminate between Hanwoo meat and other black-haired ones.

Genetic Diversity and Phylogenetic Relationship in Korean Strains of Lentinus lepideus Based on PCR Polymorphism (PCR 다형성 분석에 의한 한국산 잣버섯의 유전적 다양성 및 유연관계)

  • Lee, Jae-Seong;Cho, Hae-Jin;Yoon, Ki-Nam;Alam, Nuhu;Lee, Kyung-Lim;Shim, Mi-Ja;Lee, Min-Woong;Lee, Yun-Hae;Jang, Myoung-Jun;Ju, Young-Chul;Cheong, Jong-Chun;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, U-Youn;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.105-111
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    • 2010
  • Lentinus lepideus, known as train wrecker fungus, has been used for nutritional and medicinal purposes. Recently, commercial cultivation technique and a new cultivar of the mushroom were developed. To investigate the genetic diversity and phylogenetic relationship for identifying the mushroom strains and cultivar, one commercial and 13 strains of Lentinus lepideus from different geographical regions of Korea were analyzed by ITS regions of rDNA and RAPD of genomic DNA. Three strains of Lentinus edodes were also used for the analysis. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 173 to 179 bp and 203 to 205 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical with 156 base pairs. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into four clusters, while 3 strains of L. edodes was divided into a new cluster. Ten primers out of 20 arbitrary primers used in the RAPD-PCR efficiently amplified the genomic DNA. The numbers of amplified DNA bands varied with the primers and strains, with polymorphic DNA fragments in the range from 0.2 to 2.6 kb. The results showed that phylogenetic relationship among Korean strains of Lentnus lepideus is high, but genetic diversity is low.

Performance Comparison of Keyword Extraction Methods for Web Document Cluster using Suffix Tree Clustering (Suffix Tree를 이용한 웹 문서 클러스터의 제목 생성 방법 성능 비교)

  • 염기종;권영식
    • Proceedings of the Korea Inteligent Information System Society Conference
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    • 2002.11a
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    • pp.328-335
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    • 2002
  • 최근 들어 인터넷 기술의 발달로 웹 상에 많은 자료들이 산재해 있습니다. 사용자가 원하는 정보를 검색하기 위해서 키워드 검색을 이용하고 있는데 이러한 키워드 검색은 사용자들이 입력한 단편적인 정보에 바탕하여 검색하고 검색된 결과들을 자체적인 기준으로 순위를 매겨 나열식으로 제시하고 있다. 이러한 경우 사용자들의 생각과는 다르게 결과가 제시될 수 있다. 따라서 사용자들의 검색 시간을 줄이고 편리하게 검색하기 위한 환경의 필요성이 높아지고 있다. 본 논문에서는 Suffix Tree 알고리즘을 사용하여 관련있는 문서들을 분류하고 각각의 분류된 클러스터에 제목을 생성하기 위하여 문서 빈도수, 단어 빈도수와 역문서 빈도수, 카이 검정, 공통 정보, 엔트로피 방법을 비교 평가하여 제목을 생성하는데 어떠한 방법이 가장 효과적인지 알아보기 위해 비교 평가해본 결과 문서빈도수가 TF-IDF보다 약 10%정도 성능이 좋은 결과를 보여주었다.

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Characterization of Bovine Lymphocyte Antigen DRB3 exon2 Gene of Korean Native Cattle (한우의 BoLA DRB3 exon2 유전자의 특성)

  • Kang, Ho Bum;Ryoo, Seung Heui;Lee, Sang Hoon;Jeon, Byung Soon;Sang, Byung Chan
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.79-88
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    • 1998
  • This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.

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Comparative Molecular Phylogenetic Relationships in Different Strains of Pleurotus spp. (느타리속 버섯 계통의 분자생물학적 유연관계의 비교연구)

  • Cho, Hae-Jin;Lee, Jae-Seong;Yoon, Ki-Nam;Alam, Nuhu;Lee, Kyung-Lim;Shim, Mi-Ja;Lee, Min-Woong;Cheong, Jong-Chun;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, U-Youn;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.112-119
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    • 2010
  • Pleurotus spp. have been used for edible and medicinal purposes in Asian countries for a long time. The fruiting bodies of the Pleurotus ostreatus, Pleurotus citrinopileatus and Pleurotus salmoneostramineus contained many physiologically beneficial substances for human health. Therefore, it is necessary to study the genetic diversity of Pleurotus mushroom cultivars commercially cultivated in Korea. Eleven strains of Pleurotus spp. were collected from different geographical regions in South-East Asia and ITS regions of rDNA and RAPD of genomic DNA were analyzed. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 167 to 254 bp and 156 to 213 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into 4 clusters. Eleven Pleurotus species were also analyzed by RAPD with 20 arbitrary primers. Ten of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.1 to 2.0kb. The results revealed that genetic diversity of selected strains of P. ostreatus, P. citrinopileatus and P. salmoneostramineus is low.

Molecular Cloning and Characterization of the secY Homolog from Streptomyces lividans TK24 (Streptomyces lividans Tk24에서 secY homolog의 클로닝과 분석)

  • 김순옥;서주원
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.110-116
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    • 1998
  • The secY gene of Streptomyces lividans TK24 was cloned by the PCR method with synthetic oligonucleotide primers designed on the basis of the conserved regions of Ll5-secY-adk operon from E. coli, B. subtilis, and M luteus. The deduced amino acid sequences of the SecY are highly homologous to those of other known SecY. It has 46%, 43%, 57%, 44%, 42%,56%, 90% similarity to Escherichia coli, Bacillus subtilis, Micrococcus luteus, Bacillus licheniformis Staphylococcus carnosus, Brevibacterium flavum, Streptomyces scabies, respectively and almost the same with Streptomyces coelicolor, The gene organization of Ll5- SecY-Adk is also similar to those of other bacteria. SecY and Adk are very likely translationally coupled that is overlapping stop codon of SecY and start codon of Adk with one base pair, which is common structure among high GC content strains of gram positive bacteria.

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Organization of Antibiotic Resistance Gene Cluster of Multi-Drug Plasmid in Clinically Isolated Salmonella Enteritidis Strain (식중독환자에서 분리한 Salmonella Enteritidis 다제내성 플라스미드의 내성유전자 집락의 구조해석)

  • Jung, Seo-Yeon;Son, Chang-Kyu;Kwak Kyung-Tak;Kim, Byung-Chun;Park, Wan
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.299-305
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    • 2002
  • Clinically isolated Salmonella Enteritidis strain has a multi drug resistance plasmid, which confers ampicillin, chloramphe-nicol, sulfonamide, streptomycin and tetracycline, named pCAST2. We cloned a 7 kb Sacl fragment of pCAST2 which has sulfonamide, streptomycin and tetracycline resistance genes. The 7 kb SacI fragment showed the organization of sulII-strA-strB-tetR-tetA gene cluster which is different from the other clusters reported previously. In this study, we presented the method to detect this cluster by PCR analysis and showed that this cluster was found in Salmonella strains occurred sporadically at Kyungpook province in 2002.