• Title/Summary/Keyword: restriction mapping

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Genome Mapping of an Extreme Thermophile, Thermus caldophilus GK24

  • Park, Jong Hoon;Park, Byung Chul;Koch, Suk Hoon;Kim, Joong Soo;Koh, Jeong Heon;Yang, Moon Hee;Kim, Yong Sung;Kim, Cheorl Ho;Kim, Myoung Hee;Kwon, Suk Tae;Lee, Dae-Sil
    • Genomics & Informatics
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    • v.1 no.1
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    • pp.50-54
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    • 2003
  • Genome of an extreme thermophile, Thermus caldophilus GK24 has been analyzed to construct the genomic map. The genomic DNAs encapsulated in agarose gel were digested with SspI, EcoRI, SpeI, and HpaI restriction endonucleases, and then the resulting genomic DNA fragments were analyzed by pulsed-field gel electrophoresis. Its restriction map has been constructed by analyzing sizes of the restriction fragments obtained from both complete and partial digestions. The circular form of its genome was composed of about 1.98 Mbp and a megaplasmid. The genomic loci for the genes of xylose isomerase, thioredoxin, tRNA-16S rRNA, 23S rRNA, L5 ribosomal protein, ADP-glucose pyrophosphorylase, DNA-ligase, and Tca DNA polymerase were determined by both Southern hybridization and PCR.

Molecular Cloning of ATPase $\alpha$-Subunit Gene from Mitochondria of Korean Ginseng (Panu ginseng C.A. Meyer) (고려인삼(Panax ginseng C.A. Meyer) ATPase $\alpha$-subunit 유전자의 Cloning)

  • Park, Ui-Sun;Choi, Kwan-Sam;Kim, Kab-Sig;Kim, Nam-Won;Choi, Kwang-Tae
    • Journal of Ginseng Research
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    • v.19 no.1
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    • pp.56-61
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    • 1995
  • Molecular cloning and restriction mapping on ATPase $\alpha$-subunit gene (atpA) were carried out to obtain genomic information concerned with the gene structure and organization in Korean ginseng mitochondria. Two different clones containing the homologous sequence of atpA gene were selected from SalI and PstI libraries of mitochondrial DNA (mtDNA) of Korean ginseng. The sizes of mtDNA fragments inserted in SalI and PstI clones were 3.4 kb and 13 kb, respectively. Southern blot analysis with [$^{32}P$] labelled Oenothera atPA gene probe showed that atpA gene sequence was located in 2.0 kb XkaI fragment in PstI clone and in 1.7 kb XbaI fragment in SalI clone. A partial sequening ascertained that the SalI clone included about 1.2 kb fragment from SalI restriction site to C-terminal sequence of this gene but about 0.3 kb N-terminal sequence of open reading frame was abscent. The PstI fragment was enough large to cover the full sequence of atpA gene. The same restriction pattern of the overlapped region suggests that both clones include the same fragment of atiA locus. Data of Southern blot analysis and partial nucleotide sequencing suggested that mtDNA of Korean ginseng has a single copy of atpA gene. Key words ATPase a-subunit, mitochondrial DNA, Panax ginseng.

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Microsatellite Sequences of Mammals and Their Applications in Genome Analysis in Pigs - A Review

  • Behl, Rahul;Sheoran, Neelam;Behl, Jyotsna;Tantia, M.S.;Vijh, R.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1822-1830
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    • 2002
  • The microsatellites are the short tandem repeats of 1 to 6 bp long monomer sequences that are repeated several times. These short tandem repeats are considered to be generated by the slipped strand mispairing. Based on the unique capability of alternating purine-pyrimidine residues to form Z-DNA, the possible role of the microsatellites in gene regulation has been proposed. The microsatellites are highly polymorphic, follow Mendelian inheritance and are evenly distributed throughout the genomes of eukaryotes. They are easy to isolate and the polymerase chain reaction based typing of the alleles can be readily automated. These properties make them the preferred markers for comparison of the genetic structure of the closely related breeds/populations; very high-resolution genetic mapping and parentage testing etc. The microsatellites have rapidly replaced the restriction fragment length polymorphism (RFLP) and the random amplified polymorphic DNA (RAPD) in most applications in the population genetics studies in most species, including the various farm animals viz. cattle, buffalo, goat, sheep and pigs etc. More and more reports are now available describing the use of microsatellites in pigs ranging from measurement of genetic variation between breeds/populations, developing high resolution genetic maps to identifying and mapping genes of biological and economic importance.

Genome Mapping Technology And Its Application In Plant Breeding (작물 육종에서 분자유전자 지도의 이용)

  • 은무영
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.07a
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    • pp.57-86
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    • 1995
  • Molecular mapping of plant genomes has progressed rapidly since Bostein et al.(1980) introduced the idea of constructing linkage maps of human genome based on restriction fragment length polymorphism (RFLP) markers. In recent years, the development of protein and DNA markers has stimulated interest for the new approaches to plant improvement. While classical maps based on morphological mutant markers have provided important insights into the plant genetics and cytology, the molecular maps based on molecular markers have a number of inherent advatages over classical genetic maps for the applications in genetic studies and/or breeding schemes. Isozymes and DNA markers are numerous, discrete, non-deleterious, codominant, and almost entirely free of environmental and epistatic interactions. For these reasons, they are widely used in constructing detailed linkage maps in a number of plant species. Plant breeders improve crops by selecting plants with desirable phenotypes. However a plant's phenotyes is often under genetic control, positioning at different "quantitative trait loci" (QTLs) together with environmental effects. Molecular maps provide a possible way to determine the effect of the individual gene that combines to produce a quantitative trait because the segregation of a large number of markers can be followed in a single genetic cross. Using market-assisted selection, plants that contain several favorable genes for the trait and do not contain unfavourable segments can be obtained during early breeding processes. Providing molecular maps are available, valuable data relevant to the taxonomic relationships and chromosome evolution can be accumulated by comparative mapping and also the structural relationships between linkage map and physical map can be identified by cDNA sequencing. After constructing high density maps, it will be possible to clone genes, whose products are unknown, such as semidwarf and disease resistance genes. However, much attention has to be paid to level-up the basic knowledge of genetics, physiology, biochemistry, plant pathology, entomology, microbiology, and so on. It must also be kept in mind that scientists in various fields will have to make another take off by intensive cooperation together for early integration and utilization of these newly emerging high-techs in practical breeding. breeding.

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Molecular Characterization and Chromosomal Mapping of the Porcine AMP-activated Protein Kinase ${\alpha}2$ (PRKAA2) Gene

  • Lee, Hae-Young;Choi, Bong-Hwan;Lee, Jung-Sim;Jang, Gul-Won;Lee, Kyung-Tai;Chung, Ho-Young;Jeon, Jin-Tea;Cho, Byung-Wook;Lee, Jun-Heon;Kim, Tae-Hun
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.615-621
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    • 2007
  • AMP-activated protein kinase alpha 2 (PRKAA2) plays a key role in regulation of fatty acid and cholesterol metabolism. This study investigated the porcine PRKAA2 gene as a positional candidate for intramuscular fat and backfat thickness traits in pig chromosome 6. A partial fragment of the porcine PRKAA2 gene, amplified by PCR, contained a putative intron 3 including a part of exon 3 and 4, comparable with that of human PRKAA2 gene. Within the fragment, several single nucleotide polymorphisms were identified using multiple sequence alignments. Of these, TaqI restriction enzyme polymorphism was used for genotyping various pig breeds including Korean reference family. Using linkage and physical mapping, the porcine PRKAA2 gene was mapped in the region between microsatellite markers SW1881 and SW1680 on chromosome 6. Allele frequencies were quite different among pig breeds. The full length cDNA of the porcine PRKAA2 (2,145 bp) obtained by RACE containing 1,656 bp open reading frame of deduced 552 amino acids, had sequence identities with PRKAA2 of human (98.2%), rat (97.8%), and mouse (97.5%). These results suggested that the porcine PRKAA2 is a positional candidate gene for fat deposition trait at near telomeric region of the long arm of SSC 6.

Cloning of a Alkaline Protease Gene from Xanthomonas sp. YL-37 (Xanthomonas sp. YL-37의 Alkaline Protease 유전자의 클로닝)

  • 이대희;김수경;이승철;윤병대;황용일
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.145-149
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    • 1995
  • For the purpose of developing a new biodegradable detergent, we have isolated a gene encoding wide-range temperature applicable alkaline protease from Xanthomonas sp. YL-37 (Lee et al., 1994, Kor. J. Appl. Microbiol. Biotechnol.). An alkaline protease gene was isolated from the gene bank that was prepared from the chromosomal DNA of Xanthomonas sp. YL-37. From the results of agarose gel electrophoresis and a restriction enzyme mapping, a 2.7 kb DNA fragment containing the alkaline protease gene was inserted in the plasmid pUC9. Extracellular activity of a clone having alkaline protease gene was detected on SDS-polyacrylamide gel with activity staining assay. The molecular weight of alkaline protease was determined to be about 64 kDa from 11% SDS-PAGE analysis. Alkaline protease activity, produced from E. coli which harboring the plasmid, showed no difference at reaction temperature 20, 30 and 40$\circ$C, respectively. This result showed that alkaline protease produced from E. coli harboring the plasmid was apparently the same as that of Xanthomonas sp. YL-37.

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Molecular Cloning and Expression of a Xylanase Gene from Alkalophilic Bacillus sp.

  • Yu, Ju-Hyun;Kang, Yun-Sook;Park, Young-Seo;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.251-255
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    • 1991
  • A 16 kilobase (kb) HindIII fragment of alkalophilic Bacillus sp. YC-335 containing a gene for xylanase synthesis was inserted at the HindIII site of pBR322 and cloned in Escherichia coli HB101. After subcloning of recombinant plasmid pYS52, the 1.5 kb fragment was found to code for xylanase activity, and the hybrid plasmid was named pYS55. The DNA insert of the plasmid was subjected to restriction enzyme mapping, which showed that pYS55 had single site for PuvII and SstI in the 1.5 kb insert fragment. Southern hybridization analysis revealed that the cloned gene was hybridized with chromosomal DNA from alkalophilic Bacillus sp. YC-335. About 64% of the enzyme activity was observed in the extracellular and periplasmic space of E. coli HB10l carrying pYS55.

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Isolation and partial haracterization of rat LDH A-genomic sequences

  • Lee, Mi-Young;Yim, Sun-Young;Lee, Seung-Ki
    • Archives of Pharmacal Research
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    • v.13 no.1
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    • pp.69-73
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    • 1990
  • As a part of the study to elucidate the mechanism by which transcription of LDH A-gene is regulated by cAMP, we aimed to isolated rat LDH A gene and characterize cAMP-reponsive element (CRE). We have screened $1.2{\times}10^6$ recombinant phages of rat Charon 4A genomic library and isolated 33 positive clones among which we identified 12 different LDH A gene-related clones. By the results of restriction enzyme mapping, Southern blotting, and nucleotide sequence analyses, we concluded that the 12 LDH A gene-related clones were intronless and frequently mutated LDH A-pseudogenes. In this report, we present the characteristic features of the 12 rat liver LDH A-pseudogenes.

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Isolation of Proteinase Inhibitor II Genes from Potato (감자로부터 단백질분해효소 억제제 II 유전자의 분리)

  • 이종섭
    • Journal of Plant Biology
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    • v.32 no.2
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    • pp.79-87
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    • 1989
  • Southern hybridization of genomic DNAs with radioactively labeled cDNA of tomato proteinase inhibitor II revealed that proteinase inhibitor II proteins in potato plants are encoded by a family of about 10 related sequences. Screening of potato EcoRI genomic library with the cDNA resulted in isolation of 13 recombinant phage clones which carry 3 different genomic regions. Of these clones, clones 8, 18, and 39 were subjected to restriction mapping and subcloning. Further characterization of the subclones of clones 8, 18 and 39 indicated that two inhibitor II genes are present on a 8.0 kb EcoRI fragment of clone 8, one on 3.3 and 0.8 kb EcoRI fragments of clone 18 and two genes on a 13.5 kb EcoRI fragment of clone 39.

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Automatic Mapping of Korean Wordnet "KorLex" to Semantic Classes of Sejong Dictionary (세종 의미 부류와 KorLex 명사 어휘 의미망 자동 맵핑)

  • So, Gilja;Yoon, Aesun;Kwon, Hyuk-Chul
    • Annual Conference on Human and Language Technology
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    • 2009.10a
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    • pp.92-96
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    • 2009
  • 인간이 가진 개념을 지식베이스화하려는 시도 중 하나로 의미망이 구축되고 있다. 한국어를 대상으로 한 어휘 의미망 중 프린스턴 대학의 WordNet을 대역한 KorLex는 1,2단계에서 한국어 어휘의미의 특성을 반영하여 개념 및 의미구조를 재구조화하고 있다. 그러나 현재 KorLex의 동의어 집합을 구성하는 어휘 의미에는 논항정보를 따로 구성할 수 없었다. 본 연구는 세종 전자 사전 격틀정보내의 선택제약조건(selectional restriction)으로 사용되고 있는 의미 부류와 KorLex의 명사 어휘 의미망을 자동 맵핑하는 방안을 제안함으로써 KorLex에서 세종 전자 사전 격틀정보를 활용할 수 있는 가능성을 제공한다.

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