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ITS Primers with Enhanced Specificity to Detect the Ectomycorrhizal Fungi in the Roots of Wood Plants

  • Kim, Dong-Hun;Chung, Hung-Chae;Ohga, Shoji;Lee, Sang-Sun
    • Mycobiology
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    • v.31 no.1
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    • pp.23-31
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    • 2003
  • With universal primer ITS1-F, the specific DHJ2 primer was developed to detect the Ectomycorrhizal(ECM) root tips in soil and to identify the species of ECM fungi, as based on DNA sequences of rDNA stored in GeneBank of NCBI. This primer was designed with the common sites of rDNA of Amanita and Boletus, and was also designed with several DNA programs provided by NCBI. The DNA fragments synthesized by PCR were calculated to be 1,000 to 1,200 bps of DNA located to 18s to 28s rDNA to contain two variable sites of ITS, indicating much diversities for specific species or ecotypes of ECM fungi. The primer DHJ2 reacted with the genomic DNA's extracted from the tissues of basidiocarp at the rate of 73 of 80 fungi collected produced single bands with a 1,100 bps length. The DNA fragment synthesized with the genomic DNA that extracted from eight ECM tips of Pinus densiflora was confirmed and analysized to the rDNAs of ECM in full sequences, and informed to be a ECM fungal species in the forest.

Specific Detection of Xanthomonas oryzae pv. oryzicola in Infected Rice Plant by Use of PCR Assay Targeting a Membrane Fusion Protein Gene

  • Kang, Man-Jung;Shim, Jae-Kyung;Cho, Min-Seok;Seol, Young-Joo;Hahn, Jang-Ho;Hwang, Duk-Ju;Park, Dong-Suk
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1492-1495
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    • 2008
  • Successful control of Xanthomonas oryzae pv. oryzicola, the causal agent of bacterial leaf streak, requires a specific and reliable diagnostic tool. A pathovar-specific PCR assay was developed for the rapid and accurate detection ofthe plant pathogenic bacterium Xanthomonas oryzae pv. oryzicola in diseased plant. Based on differences in a membrane fusion protein gene of Xanthomonas oryzae pv. oryzicola and other microorganisms, which was generated from NCBI (http://www.ncbi.nlm.nih.gov/) and CMR (http://cmr.tigr.org/) BLAST searches, one pair of pathovar-specific primers, XOCMF/XOCMR, was synthesized. Primers XOCMF and XOCMR from a membrane fusion protein gene were used to amplity a 488-bp DNA fragment. The PCR product was only produced from 4 isolates of Xanthomonas oryzae pv. oryzicola among 37 isolates of other pathovars and species of Xanthomonas, Pectobacterium, Pseudomonas, Burkholderia, Escherichia coli, and Fusarium oxysporum f.sp. dianthi. The results suggested that the assay detected the pathogen more rapidly and accurately than standard isolation methods.

Spermatogonia 단계에 특이적으로 발현하는 유전자 동정

  • 옥도원;김진회
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.48-48
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    • 2003
  • 본 실험은 spermatogonia 단계에 발현하는 유전자를 찾기 위하여 suppression subtractive hybridization를 수행하였다. 기존에 mouse에서는 spermatogonia 특이적인 유전자들이 밝혀져 있기 때문에 pig에 특이적인 유전자를 찾기 위하여 pig 250days testis와 pig 60days testis를 재료로 하여 실험하였다. SSH를 통하여 254days testis에 특이적으로 발현되는 후보유전자를 7개 찾았고 25days testis와 60days testis 의 Northern blot을 통하여 25days에 과발현하고 60days에 발현의 양이 대폭 줄어드는 spermatogonia 유전자로 생각되는 후보유전자 2개를 선택하여 pig tissue northern blot, genomic DNA southern blot, RT-PCR 그리고 In-situ hybridization을 수행하였다. Tissue northern blot과 RT-PCR을 통하여 후보자 1번은 간과 폐, 난소, 정소에서 발현하고, 후보유전자 15번은 난소와 정소에서만 특이적으로 발현함을 알았다. DNA sequence analysis와 NCBI Blast search를 통하여 후보자 1번은 다른 종에서 밝혀진 유전자였고 후보유전자 15번은 어느 종에서도 밝혀지지 않은 새로운 유전자였다. Degenerated primer를 통하여 후보자 1번의 pig full sequence를 밝히고 NCBI에 등록하였다. 그리고 In-situ hybridization을 통하여 후보유전자득이 20일째 testis의 Leydic cell에서 많이 발현되고 adult testis에서는 발현이 감소하는 결과를 얻었다. 이것으로 보아 위의 두 후보유전자는 spermatogonia에 직접 관련된 유전자이기 보다는 spermatogonia의 발달에 영향을 주는 leydic cell 특이발현을 가진 유전자로 사료되어진다.

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Genomic and evolutionary analysis with gluten proteins of major food crops in the Triticeae tribe

  • Kim, Sang Heon;Seo, Yong Weon
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.86-86
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    • 2017
  • Prolamins are the main seed storage proteins in cereals. Gluten proteins seem to be prolamins because their primary structure have the meaningful quantity of proline and glutamine amino acid residues. Gluten proteins are found in crops such as wheat (Triticum aestivum), barley (Hordeum vulgare), and rye (Secale cereale) which are major food crops in the Triticeae tribe. Glutenin and gliadin, hordein, and secalin are typical gluten proteins found in wheat, barley, and rye, respectively. Gluten affect grain quality so that many researches, such as isolation or characterization of their genes, have been carried out. To improve the quality of grains in the Triticeae tribe, it is necessary to understand the relationship within their gluten proteins and their evolutionary changes. The sequences of nucleotides and amino acids of gluten protein including glutenins, gliadins, hordeins, and secalins were retrieved from NCBI (https://www.ncbi.nlm.nih.gov/) and Uniprot (http://www.uniprot.org/). The sequence analysis and the phylogenetic analysis of gluten proteins were performed with various website tools. The results demonstrated that gluten proteins were grouped with their homology and were mostly corresponded with the previous reports. However, some genes were moved, duplicated, or disappeared as evolutionary process. The obtained data will encourage the breeding programs of wheat, barley, rye, and other crops in the Triticeae tribe.

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GTVseq: A Web-based Genotyping Tool for Viral Sequences

  • Shin, Jae-Min;Park, Ho-Eun;Ahn, Yong-Ju;Cho, Doo-Ho;Kim, Ji-Han;Kee, Mee-Kyung;Kim, Sung-Soon;Lee, Joo-Shil;Kim, Sang-Soo
    • Genomics & Informatics
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    • v.6 no.1
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    • pp.54-58
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    • 2008
  • Genotyping Tool for Viral SEQuences (GTVseq) provides scientists with the genotype information on the viral genome sequences including HIV-1, HIV-2, HBV, HCV, HTLV-1, HTLV-2, poliovirus, enterovirus, flavivirus, Hantavirus, and rotavirus. GTVseq produces alternative and additive genotype information for the query viral sequences based on two different, but related, scoring methods. The genotype information produced is reported in a graphical manner for the reference genotype matches and each graphical output is linked to the detailed sequence alignments between the query and the matched reference sequences. GTVseq also reports the potential 'repeats' and/or 'recombination' sequence region in a separated window. GTVseq does not replace completely other well-known genotyping tools such as NCBI's virus sequence genotyping tool (http://www.ncbi. nlm.nih.gov/projects/genotyping/formpage.cgi), but provides additional information useful in the confirmation or for further investigation of the genotype(s) for the newly isolated viral sequences.

Screening of Fruiting Body Formation-Specific Genes from the Medicinal Mushroom Cordyceps militaris MET7903 (약용버섯번데기 동충하초 MET7903의 특이적 자실체형성 유전자의 선별)

  • Yun, Bangung;Chung, Ki-Chul
    • Journal of Mushroom
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    • v.2 no.3
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    • pp.145-148
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    • 2004
  • This study was carried out to screen the fruiting body formation-specific genes from the medicinal mushroom Cordyceps militaris. A cDNA synthesized using total RNA from 4 stages of mushroom development, mycelium, primordium, immature fruiting body and mature fruiting body. Differential expression gene screening was performed by DD-PCR(Differential Display Arbitrary Primer PCR) with cDNA, we sequenced partial 6 genes using pGEM cloning vector. The DNA Sequence of the six DD-PCR products derived from differentially expressed genes was compared to that in the GenBank database by using the NCBI BLAST search to identify similarities to known sequences. Sequence analysis showed that six of DD-PCR products have unknown sequence.

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IntoPub: A Directory Server for Bioinformatics Tools and Databases

  • Jung, Dong-Soo;Kim, Ji-Han;Lee, Sang-Hyuk;Lee, Byung-Wook
    • Interdisciplinary Bio Central
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    • v.3 no.3
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    • pp.12.1-12.3
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    • 2011
  • Bioinformatics tools and databases are useful for understanding and processing various biological data. Numerous resources are being published each year. It is not a trivial task to find up-to-date relevant tools and databases. Moreover, no server is available to provide comprehensive coverage on bioinformatics resources in all biological fields. Here, we present a directory server called IntoPub that provides information on web resources. First, we downloaded XML-formatted abstracts containing web URLs from the NCBI PubMed database by using 'ESearch-EFetch' function in the NCBI E-utilities. The information is obtained from abstracts in the PubMed by extracting 'www' or 'http' prefixes. Then, we cu-rate the downloaded abstracts both in automatic and manual fashion. As of July 2011, the IntoPub database has 12,118 abstracts containing web URLs from 174 journals. Our anal-ysis shows that the number of abstracts containing web resources has increased signifi-cantly every year. The server has been tested by many biologists from several countries to get opinion on user satisfaction, usefulness, practicability, and ease of use since January 2010. In the IntoPub web server, users can easily find relevant bioinformatics resources, as compared to searching in PubMed. IntoPub will continue to update and incorporate new web resources from PubMed and other literature databases. IntoPub, available at http://into.kobic.re.kr/, is updated every day.

Construction of the Aging Related Gene Database using Text-mining (서지분석을 통한 노화 관련 유전자 정보 데이터베이스 구축)

  • Yu, Seok Jong;Park, Junho;Yoo, Jaesoo
    • Proceedings of the Korea Contents Association Conference
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    • 2013.05a
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    • pp.41-42
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    • 2013
  • 최근 노령화가 급속히 진행되면서 노화에 대한 연구가 활발히 진행되고 있다. 하지만 노화현상은 광범위한 표현형을 지니고 있는 생명현상으로 이에 대한 체계적인 연구를 지원하기 위한 웹포털 사이트가 필요한 실정이다. 특히 노화에 따른 질병과의 연관성 및 관련 유전자에 대한 정보를 수집하고 이를 체계적으로 분석할 수 있는 통합정보시스템은 향후 노화연구를 지원하기 위한 가장 핵심적인 요소라고 할 수 있다. 본 연구에서는 기존 노화와 관련된 461개의 유전자를 기반으로 관련된 질병과의 연관성을 OMIM 데이터베이스를 활용하여 분석하였다. 또한 관련 단백질의 기능을 GO데이터베이스 분석을 통해 유전자의 기능을 분석하였다. Pubmed에서 제공하는 노화관련 논문들의 MeSH 정보 분석을 통해서 노화와 관련된 용어를 분석하였다. 노화와 관련된 64개의 유전자를 키워드로 NCBI의 pubmed 데이터베이스로부터 관련논문을 다운로드 받아 생물학적 상호작용 정보를 추출했다. 생물학적 상호작용은 NCBI에서 제공하는 Metamap 데이터베이스를 기반으로 각각의 생물학적 용어를 정의했다. 현재 노화 유전자 64개에 대해 128,729개의 생물학적 상호작용 정보를 추출했고, 8대 노인성만성질환에 대해 301,176개의 생물학적 상호작용 정보를 추출하였다.

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Screening of Genes Which are Able to Affect Kalanchoe Vegetative Reproduction (Kalanchoe 식물의 영양 번식에 영향을 줄 수 있는 유전자들의 선발)

  • Jung, Yu-Chul;Chung, Young-Jae;Kim, Dong-Giun
    • Journal of Life Science
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    • v.21 no.6
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    • pp.865-874
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    • 2011
  • The genus Bryophyllum is best known for many of its species having the ability to produce plantlets on their leaves. This phenomenon is also known as vegetative reproduction. Differential expressed gene (DEG) detecting technique was applied in order to survey the genes involved in the process of asexual reproduction for plantlet formation. Based on homology search using the NCBI database after screening of genes, 38 genes were identified from a total of 69 DEGs. Most of these DEGs were related to cell division, to intercellular signal transduction, and to hormone (cytokinin and ethylene) signaling.