• Title/Summary/Keyword: Genomic hybridization

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Quantitative analysis using decreasing amounts of genomic DNA to assess the performance of the oligo CGH microarray

  • Song Sunny;Lazar Vladimir;Witte Anniek De;Ilsley Diane
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2006.02a
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    • pp.71-76
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    • 2006
  • Comparative genomic hybridization (CGH) is a technique for studying chromosomal changes in cancer. As cancerous cells multiply, they can undergo dramatic chromosomal changes, including chromosome loss, duplication, and the translocation of DNA from one chromosome to another. Chromosome aberrations have previously been detected using optical imaging of whole chromosomes, a technique with limited sensitivity, resolution, quantification, and throughput. Efforts in recent years to use microarrays to overcome these limitations have been hampered by inadequate sensitivity, specificity and flexibility of the microarray systems. The oligonucleotide CGH microarray system overcomes several scientific hurdles that have impeded comparative genomic studies of cancer. This new system can reliably detect single copy deletions in chromosomes. The system includes a whole human genome microarray, reagents for sample preparation, an optimized microarray processing protocol, and software for data analysis and visualization. In this study, we determined the sensitivity, accuracy and reproducibility of the new system. Using this assay, we find that the performance of the complete system was maintained over a range of input genomic DNA from 5 ug down to 0.15 ug.

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Genomic Organization of Penicillium chrysogenum chs4, a Class III Chitin Synthase Gene

  • Park, Yoon-Dong;Lee, Myung-Sook;Kim, Ji-Hoon;Jun Namgung;Park, Bum-Chan;Bae, Kyung-Sook;Park, Hee-Moon
    • Journal of Microbiology
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    • v.38 no.4
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    • pp.230-238
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    • 2000
  • Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5'flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

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High Resolution Genomic Profile of Neuro2a Murine Neuroblastoma Cell Line by Array-based Comparative Genomic Hybridization (고집적어레이 기반의 비교유전체보합법(CGH)을 통한 신경아세포종 Neuro2a 세포의 유전체이상 분석)

  • Do, Jin-Hwan;Kim, In-Su;Ko, Hyun-Myung;Choi, Dong-Kug
    • Journal of Life Science
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    • v.19 no.4
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    • pp.449-456
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    • 2009
  • Murine Neuro-2a (N2a) cells have been widely used for the investigation of neuronal differentiation, trophic interaction and neurotoxic effects of various compounds and their associated mechanisms. N2a cells have many genomic variations such as gains or losses in DNA copy number, similar to other neuroblastoma cells, and no systematic or high-resolution studies of their genome-wide chromosomal aberrations have been reported. Presently, we conducted a systematic genome-wide determination of chromosomal aberrations in N2a cells using a high-throughput, oligonucleotide array-based comparative genomic hybridization (oaCGH) technique. A hidden Markov Model was employed to assign each genomic oligonucleotide to a DNA copy number state: double loss, single loss, normal, gain, double gain and amplification. Unlike most neuroblastoma cells, Mycn amplification was not observed in N2a cells. In addition, these cells showed gain only in the neuron-derived neurotrophic factor (NF), while other neurotrophic factors such as glial line-derived NF and brain-derived NF presented normal copy numbers. Chromosomes 4, 8, 10, 11 and 15 displayed more than 1000 aberrational oligonucleotides, while chromosomes 3, 17, 18 and 19 displayed less than 20. The largest region of gain was located on chromosome 8 and its size was no less than 26.7 Mb (Chr8:8427841-35162415), while chromosome 4 had the longest region of single deletion, with a size of 15.1 Mb (Chr4:73265785-88374165).

Development of Repetitive DNA Probes for Genetic Analysis of Phytophthora capsici (Phytophthora capsici의 유전적 특성 분석을 위한 Repetitive DNA Probe의 개발)

  • Song, Jeong-Young;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.66-72
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    • 2002
  • To develop DNA markers for analysis of genetic characteristics of Phytophthora capsici population, randomly selected clones from HindIII-digested genomic DNA library of P. capsici 95CY3119 were surveyed by hybridizing to Southern blots of HindIII-digested total genomic DNA of P. capsici. Probe DNAs inserted into selected individual clones strongly hybridized with HindIII digests of P. capsici. Among probes examined, PC9 revealed the repetitive and highly polymorphic bands to HindIII digests of inter-and intra-field P. capsici isolates. Genetic diversity of individual isolates was also clearly revealed in cluster analysis based on its band patterns. The other probe, PC22, was hybridized only to DNA from P. capsici and this was highly repetitive. However, there was no response to other Phytophthora species and Pythium sp. These DNA probes could be used as very useful markers in analysing genetic diversity and identification for P. capsici population throughout the world.

Actinodura roseorufa에서 생산되는 UK-58,852로부터 PKS type I 에 관련된 생합성 유전자의 분리 및 분석

  • Kim, Ja-Yong;Lee, Ju-Ho;Kim, Dae-Hui;Kim, Dong-Hyeon;Song, Jae-Gyeong;Lee, Hui-Chan
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.660-664
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    • 2000
  • To clone genes related UK-58,852 production, genomic DNA of strain Actinodura roseorufa was used for the construction of genomic library using pOJ446 cosmid vector. The genomic library was screened rising dehydratase PCR product and eryA gene as a DNA hybridization probe. pHD54 was isolated, which contained an approximately 35kb of inserted DNA. BamHI, SmaI and sonicater fragments hybridized to eryA probe. All of pHD54 BgmHI, SmaI and sonicater fragments were subcloned into pGEM7 and some fragments which hybridized to eryA probe were sequenced. The nucleotide sequence was analysed using BLAST program. The sequence identities were observed in KS,AT, KR, ER and PKS loading domains. Also oxidoreductase showed similarity to rifamycin module10, and dTDP-D-glucose 4,6 dehydratase and TDP-D-glucose synthase involved in biosynthesis of sugar showed similarity to Streptomyces argillaceus.

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Genomic Alterations in Korean Laryngeal Squamous Cell Carcinoma: Array-Comparative Genomic Hybridization (한국인 후두 편평 상피 세포암의 유전체 이상분석: Array 비교 유전체 보합법)

  • Cho, Yoon-Hee;Park, Soo-Yeun;Lee, Dong-Wook;Kim, Han-Su;Lee, Ja-Hyun;Park, Hae-Sang;Chung, Sung-Min
    • Korean Journal of Head & Neck Oncology
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    • v.24 no.2
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    • pp.155-161
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    • 2008
  • Head and neck squamous cell carcinoma(HNSCC) still has poor outcome, and laryngeal cancer is the most frequent subtype of HNSCC. Therefore, there is a need to develop novel treatments to improve the outcome of patients with HNSCC. It is critical to gain further understanding on the molecular and chromosomal alteration of HNSCC to identify novel therapeutic targets but genetic etiology of squamous cell carcinoma of the larynx is so complex that target genes have not yet been clearly identified. Array based CGH(array-CGH) allows investigation of general changes in target oncogenes and tumor suppressor genes, which should, in turn, lead to a better understanding of the cancer process. In this study, We used genomic wide array-CGH in tissue specimens to map genomic alterations found in laryngeal squamous cell carcinomas. As results, gains of MAP2, EPHA3, EVI1, LOC389174, NAALADL2, USP47, CTDP1, MASP1, AHRR, and KCNQ5, with losses of SRRM1L, ANKRD19, FLJ39303, ZNF141, DSCAM, GPR27, PROK2, ARPP-21, and B3GAT1 were observed frequently in laryngeal squamous cell carcinoma tissue specimens. These data about the patterns of genomic alterations could be a basic step for understanding more detailed genetic events in the carcinogenesis and also provide information for diagnosis and treatment in laryngeal squamous cell carcinoma. The high resolution of array-CGH combined with human genome database would give a chance to find out possible target genes which were gained or lost clones.

Immunohistochemistry for the Detection of Swine hepatitis E virus in the liver

  • Ha, Seung-Kwon;Chae, Chan-hee
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2003.10a
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    • pp.28-28
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    • 2003
  • Hepatitis E virus (HEV), previously referred to as enterically transmitted non-A, non-B hepatitis, is responsible for sporadic infections as well as large epidemics of acute viral hepatitis in developing countries. The disease generally affects young adults and reportedly has a mortality rate of up to 20% in infected pregnant women. HEV was once considered to be a member of the family Caliciviridae, but the unique genomic organization of HEV has led to the removal of HEV from the family and it was provisionally classified in an unassigned family of HEV-like viruses. In situ hybridization provides any cellular detail and histological architecture.[1] However, use of in situ hybridization is largely restricted to the laboratories because this technique is the greater technical complexity and expense compared with immunohistochemistry. Therefore, the objective of this study is to develop the immunohistochemistry for the detection of swine HEV from formalin-fixed, paraffin-embedded hepatic tissues. (omitted)

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Characterization of a New Leuconostoc Species Isolated from Fresh Garlic

  • Lee, Se-Hi;Choi, Jong-Hoon;Kim, Youn-Soon;Kyung, Kyu-Hang
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.416-419
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    • 2005
  • Unknown bacterium isolated from garlic was characterized using phenotypic methods, phylogenetic analysis, DNA-DNA hybridization, and cultural methods. The strain was identified as typical leuconostoc; Gram-positive, non-sporeforming, heterofermentative, catalase-negative and spherical. Although its 16S rRNA gene sequence showed high homology to Leuconostoc argentinum DSM $8581^T$(99.8%), DNA-DNA hybridization experiments indicated it represents novel genomic species in the genus Leuconostoc. The garlic-specific leuconostoc was more resistant to antimicrobial activity of garlic compared to other common laboratory lactic acid bacteria, and was even stimulated by low concentrations (1-2%) of garlic extract supplemented in trypticase soy broth. Growth stimulation was concentration-dependent when tested with residual aqueous layer after solvent extraction of fresh whole garlic extract.

Development of PCR-microplate Hybridization Assay for Detection of Mycobacterium tuberculosis

  • Lee, In-Soo;Cho, Een-Jin;Cho, Sang-Nae;Kim, Tae-Ue;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.295-300
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    • 2009
  • Tuberculosis caused by Mycobacterium tuberculosis (MTB) still remains to be the most dreadful infectious disease affecting almost every country. In the present study, we developed a simple and rapid but accurate and sensitive assay method for detecting MTB using microplate hybridization assay. For this, a selective region of the rpoB gene was used to design PCR primers, and MTB and Mycobacterium genus-specific probe molecules. The specificity of the assay was confirmed using fifteen different mycobacterial reference strains and twelve different non-mycobacterial reference strains, and the sensitivity was determined to be 100 fg using genomic DNA of MTB reference strain, H37Rv. Subsequently, a total of 62 sputum samples with diverse smear scores and culture positive results were used to evaluate the kit performance. In brief, the specificity and the sensitivity of the assay were 100% and 98.4%, respectively.

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Development of Wheat Breeding Material Mediated wide Hybridization Response to Climate Change

  • Seong-Wook Kang;Ji-Yoon Han;Seong-Woo Cho
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.293-293
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    • 2022
  • This study is to develop new wheat breeding material through wide hybridization with wild species harboring useful characteristics such as salt, heat, and drought tolerance. Leymus mollis, wild rye was used to improve wheat genetic quality. L. mollis, is a perennial plant harboring tolerance against salt, heat, and drought because L. mollis distributes on the seaside. The F1 hybrids were produced by crossing between common wheat (Triticum aestivum L., Chinese Spring) and L. mollis. Genomic in situ hybridization revealed that the F1 hybrids have L. mollis genome. For the evaluation of salt and drought tolerance, seeds from the F2 were used. Under 2% NaCl solution, the F3 wheat-Leymus addition plants with salt tolerance showed more tillering and longer roots than other F3 plants without salt tolerance. Also, the F3 plants with salt tolerance showed better shallow-rooted than other F3 plants without salt tolerance. Finally, the F3 plants with salt tolerance made seed-setting under 2% NaCl condition, but other F3 plants without salt tolerance were not. Under drought conditions, the F3 plants with drought tolerance showed longer culm and spike length than other F3 plants without drought tolerance and even those of Chinese Spring under well-water conditions. We evaluated and selected the F3 plants with salt or drought tolerance for generation advancement.

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