• Title/Summary/Keyword: reference gene

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Studies on the pathogenicity of Salmonella typhimurium isolated from slaughtered pigs in Chonnam area (전남지역 도축돈에서 분리된 Salmonella typhimurium의 병원성에 관한 연구)

  • 정대영;박종태;고홍범
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.39-50
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    • 2003
  • Non-typhoidal Salmonella serovars remain a potential threat to human health and many animals including beef cattle, broiler chickens, and pigs which possible sources of non-typhoidal salmonellosis in human. In this study, the cecal contents of slaughtered pigs were examined for Salmonella serovar prevalence. The characteristics of the isolates, including antimicrobial resistance patterns and virulence genes, were studied along with the reference strain S typhimurium ATCC 13311. Out of 640 sample, 137 Salmonella(21.4%) were isolated and their serovar were identified S typhimurium 83 strains(60.6%), S agona 10 strains(7.3%), S schwarzengrund 4 strains(2.9%), S derby 4 strains(2.9%), S ayinde 1 strains(0.7%), and untypable 35 strains(25.5%). All 83 S typhimurium strains(100%) were multi-drug resistance to at least 7 antibiotics, and 20 strains(24.1%) of 83 isolates were R-type ACSSuT. Examination of virulent gene by PCR revealed that 73 S typimurium field isolates(88%) have a invA gene and 24 strains(28.9%) have a spvC gene. Consequently, S typhimurium infection in slaughtered pigs was relatively to appear high prevalence in their herds which suggested that it should be necessary for herd health monitoring and surveillance.

Multiplex PCR Detection for 3 Events of Genetically Modified Maize, DAS-59122-7, TC6275, and MIR604

  • Ahn, Ji-Hye;Kim, Jae-Hwan;Kim, Su-Youn;Lee, Woo-Young;Park, Sun-Hee;Kim, Hae-Yeong
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.569-572
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    • 2008
  • A multiplex polymerase chain reaction (PCR) method was developed to simultaneously detect 3 events of genetically modified (GM) maize. The event-specific primers were used to discriminate the following 3 events of GM maize (DAS-59122-7, TC6275, and MIR604) using multiplex PCR method. The zein gene was used as an endogenous maize reference gene in the multiplex PCR detection. The primer pair Zein-FIR producing a 99 bp amplicon was used to amplify the zein gene. The primer JI-Das-F1/R1 for DAS-59122-7, JI-TC6275-F3/R3 for TC6275, and JI-MIR F1/R1 for MIR604 yielded an amplicon of 130, 162, and 197 bp, respectively. The detection limit of multiplex PCR was 1% for DAS-59122-7, TC6275, and MIR604 for one reaction.

Characterization of Pseudomonas syringae pv. syringae, Causal Agent of Citrus Blast of Mandarin in Montenegro

  • Ivanovic, Zarko;Perovic, Tatjana;Popovic, Tatjana;Blagojevic, Jovana;Trkulja, Nenad;Hrncic, Snjezana
    • The Plant Pathology Journal
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    • v.33 no.1
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    • pp.21-33
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    • 2017
  • Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin (Citrus reticulata) in Montenegro, using multilocus sequence analysis of gyrB, rpoD, and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB, rpoD, and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control.

Development and Validation of the Custom Human cDNA Microarray (KISTCHIP-400) for Monitoring Expression of Genes involved in Hormone Disruption

  • Kim, Youn-Jung;Chang, Suk-Tai;Yun, Hye-Jung;Jeon, Hee-Kyung;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.180-180
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    • 2003
  • Transcript profiling is a particularly valuable tool in the field of steroid receptor biology, as these receptors are ligand-activated transcription factors and therefore exert their initial effects through altering gene expression in responsive cells. Also, an increased awareness of endocrine disrupting chemicals (EBCs) and their potential to affect wildlife and humans has produced a demand for practical screening methods to identify endocrine activity. Here we developed an in-house cDNA microarray, named KISTCHIP-400, with 401 clones, hormone related genes, factors, and ESTs, based on public database and research papers. Theses clones contained estrogen, androgen, thyroid hormone St receptors, sex hormone signal transduction & regulation, c-fos, c-myc, ps2 gene, metabolism related genes etc. And to validate the KISTCHIP-400, we investigated gene expression profiles with reference hormones, 10$\^$-8/ M 17be1a-estradiol, 10$\^$-7/ M testosterone, 10$\^$-7/ M progesterone, and thyroxin in MCF-7 cell line. Although it is in first step of validation, low doses and combinations of EDCs need to be tested. Our preliminary results that indicate the developed microarray may be a useful laboratory tool for screening EDCs and elucidating endocrine disrupting mechanism.

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Identification of Virulence Factors in Vibrio vulnificus by Comparative Transcriptomic Analyses between Clinical and Environmental Isolates Using cDNA Microarray

  • Kim, In-Hwang;Kim, Byung-Soo;Lee, Kyung-Shin;Kim, Ik-Joong;Son, Jee-Soo;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1228-1235
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    • 2011
  • We compared the gene expression among four clinical and five environmental V. vulnificus isolates, using a cDNA microarray containing 131 genes possibly associated with pathogenicity, transport, signal transduction, and gene regulations in the pathogen. cDNAs from total RNAs of these isolates were hybridized into the cDNA microarray using the cDNA of the wild-type strain MO6-24/O as a reference. We focused on selecting differentially expressed (DE) genes between clinical and environmental isolates using a modified t-statistic. We could detect two statistically significant DE genes between virulent isolates and less-virulent isolates with a marginal statistical significance (p-value of 0.008). These were genes putatively encoding pilin and adenlyate cylase. Real time-PCR confirmed that these two selected genes transcribed in significantly higher levels in virulent isolates than in less-virulent isolates. Mutants with lesions in the gene encoding pilin showed significantly higher $LD_{50}$ values than that of wild type.

Applications of Genetically Modified Tools to Safety Assessment in Drug Development

  • Kay, Hee-Yeon;Wu, Hong-Min;Lee, Seo-In;Kim, Sang-Geon
    • Toxicological Research
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    • v.26 no.1
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    • pp.1-8
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    • 2010
  • The process of new drug development consists of several stages; after identifying potential candidate compounds, preclinical studies using animal models link the laboratory and human clinical trials. Among many steps in preclinical studies, toxicology and safety assessments contribute to identify potential adverse events and provide rationale for setting the initial doses in clinical trials. Gene modulation is one of the important tools of modern biology, and is commonly employed to examine the function of genes of interest. Advances in new drug development have been achieved by exploding information on target selection and validation using genetically modified animal models as well as those of cells. In this review, a recent trend of genetically modified methods is discussed with reference to safety assessments, and the exemplary applications of gene-modulating tools to the tests in new drug development were summarized.

Development of Species-Specific Primers for PCR Identification of Lactobacillus hilgardii and Lactobacillus farciminis in Kimchi

  • Lee, Myung-Ki;Ku, Kyung-Hyung;Kim, Young-Jin;Kim, Kyung-Hee;Kim, Yu-Ri;Yang, Hye-Jung
    • Preventive Nutrition and Food Science
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    • v.15 no.2
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    • pp.159-166
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    • 2010
  • The aim of this study was to develop species-specific primer sets for kimchi Lactobacillus. Known gene sequences of Lactobacillus 16S rRNA were collected from the NCBI Gene bank, and 69 primer sets were designed using the homologous gene sequence. Six species of kimchi Lactobacilli were used as reference strains: Lactobacillus brevis KCTC3102, Lactobacillus farciminis KCTC3681, Lactobacillus fermentum KCTC3112, Lactobacillus hilgardii KCTC3500, Lactobacillus plantarum KCTC3099, and Lactobacillus sanfranciscensis KCTC3205. PCR amplification and gel electrophoresis were performed to identify the accuracy and specificity of the developed primer set. The results show that the primer set of 5'-aagcctgcgaaggcaag-3' & 5'-aggccaccggctttg-3', 5'-acatactatgcaaatctaagagattagacg-3' & 5'-actgagaatggctttaagagattagcttac-3' resulted in a specific PCR band on L. hilgardii, and primer set of 5'-ctaataccgcataacaactactttcacat-3' & 5'-aacttaataaaccgcctacattctctttac-3' on L. farciminis. The results indicate that the developed primer sets can provide a useful tool for the identification and differentiation of L. hilgardii and L. farciminis from other Lactobacillus species of kimchi.

Application of DNA Microarray Technology to Molecular Microbial Ecology

  • Cho Jae-Chang
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.22-26
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    • 2002
  • There are a number of ways in which environmental microbiology and microbial ecology will benefit from DNA micro array technology. These include community genome arrays, SSU rDNA arrays, environmental functional gene arrays, population biology arrays, and there are clearly more different applications of microarray technology that can be applied to relevant problems in environmental microbiology. Two types of the applications, bacterial identification chip and functional gene detection chip, will be presented. For the bacterial identification chip, a new approach employing random genome fragments that eliminates the disadvantages of traditional DNA-DNA hybridization is proposed to identify and type bacteria based on genomic DNA-DNA similarity. Bacterial genomes are fragmented randomly, and representative fragments are spotted on a glass slide and then hybridized to test genomes. Resulting hybridization profiles are used in statistical procedures to identify test strains. Second, the direct binding version of microarray with a different array design and hybridization scheme is proposed to quantify target genes in environmental samples. Reference DNA was employed to normalize variations in spot size and hybridization. The approach for designing quantitative microarrays and the inferred equation from this study provide a simple and convenient way to estimate the target gene concentration from the hybridization signal ratio.

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Analysis of Hemocyte-specific Gene Expression from Bombyx mori

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong-Ryul;Kang, Seok-Woo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.1
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    • pp.137-141
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    • 2011
  • A previous data was provided information for tissuespecific expression genes by means of whole-genome oligonucleotide microarray in the silkworm. We analyzed the tissue-specific expression patterns in the hemocyte tissue on 5 days of 5th instar larvae during the development of $B.$ $mori$. Total 5 candidates pick out from the $Bombyx$ $mori$ Microarray Database (BmMDB; http://silkworm.swu.edu.cn/microarray). To verify the hemocyte-specific expression, we analyzed by semi-quantitative and real-time quantitative RT-PCR using the highly expressed endogenous $Actin$ RNA as an intrinsic reference. In this study, we confirmed that one gene-sw17255- out of 5 candidates expressed in the hemocyte tissue, which was consistent with the previous data. Circulating hemocytes in the body fluid of the $B.$ $mori$ are most powerful target organ for producing biomaterials. We need further studies to find hemocyte-specific promoter region from sw17255 gene. Finally, this result can be applied in creating transgenic silkworms as a biomedical insect.

Functional Annotation and Analysis of Korean Patented Biological Sequences Using Bioinformatics

  • Lee, Byung Wook;Kim, Tae Hyung;Kim, Seon Kyu;Kim, Sang Soo;Ryu, Gee Chan;Bhak, Jong
    • Molecules and Cells
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    • v.21 no.2
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    • pp.269-275
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    • 2006
  • A recent report of the Korean Intellectual Property Office(KIPO) showed that the number of biological sequence-based patents is rapidly increasing in Korea. We present biological features of Korean patented sequences though bioinformatic analysis. The analysis is divided into two steps. The first is an annotation step in which the patented sequences were annotated with the Reference Sequence (RefSeq) database. The second is an association step in which the patented sequences were linked to genes, diseases, pathway, and biological functions. We used Entrez Gene, Online Mendelian Inheritance in Man (OMIM), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO) databases. Through the association analysis, we found that nearly 2.6% of human genes were associated with Korean patenting, compared to 20% of human genes in the U.S. patent. The association between the biological functions and the patented sequences indicated that genes whose products act as hormones on defense responses in the extra-cellular environments were the most highly targeted for patenting. The analysis data are available at http://www.patome.net