• Title/Summary/Keyword: phenotypic microarray

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A New Isolation and Evaluation Method for Marine-Derived Yeast spp. with Potential Applications in Industrial Biotechnology

  • Zaky, Abdelrahman Saleh;Greetham, Darren;Louis, Edward J.;Tucker, Greg A.;Du, Chenyu
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1891-1907
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    • 2016
  • Yeasts that are present in marine environments have evolved to survive hostile environments that are characterized by high exogenous salt content, high concentrations of inhibitory compounds, and low soluble carbon and nitrogen levels. Therefore, yeasts isolated from marine environments could have interesting characteristics for industrial applications. However, the application of marine yeast in research or industry is currently very limited owing to the lack of a suitable isolation method. Current methods for isolation suffer from fungal interference and/or low number of yeast isolates. In this paper, an efficient and non-laborious isolation method has been developed and successfully isolated large numbers of yeasts without bacterial or fungal growth. The new method includes a three-cycle enrichment step followed by an isolation step and a confirmation step. Using this method, 116 marine yeast strains were isolated from 14 marine samples collected in the UK, Egypt, and the USA. These strains were further evaluated for the utilization of fermentable sugars (glucose, xylose, mannitol, and galactose) using a phenotypic microarray assay. Seventeen strains with higher sugar utilization capacity than the reference terrestrial yeast Saccharomyces cerevisiae NCYC 2592 were selected for identification by sequencing of the ITS and D1/D2 domains. These strains belonged to six species: S. cerevisiae, Candida tropicalis, Candida viswanathii, Wickerhamomyces anomalus, Candida glabrata, and Pichia kudriavzevii. The ability of these strains for improved sugar utilization using seawater-based media was confirmed and, therefore, they could potentially be utilized in fermentations using marine biomass in seawater media, particularly for the production of bioethanol and other biochemical products.

Gene Expression Profiling in Hepatic Tissue of two Pig Breeds

  • Jang, Gul-Won;Lee, Kyung-Tai;Park, Jong Eun;Kim, Heebal;Kim, Tae-Hun;Choi, Bong-Hwan;Kim, Myung Jick;Lim, Dajeong
    • Journal of Animal Science and Technology
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    • v.54 no.6
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    • pp.383-394
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    • 2012
  • Microarray analyses provide information that can be used to enhance the efficiency of livestock production. For example, microarray profiling can potentially identify the biological processes responsible for the phenotypic characteristics of porcine liver. We performed transcriptome profiling to identify differentially expressed genes (DEGs) in liver of pigs from two breeds, the Korean native pigs (KNP) and Yorkshire pigs. We correctly identified expected DEGs using factor analysis for robust microarray summarization (FARMS) and robust multi-array average (RMA) strategies. We identified 366 DEGs in liver (p<0.05, fold-change>2). We also performed functional analyses, including gene ontology and molecular network analyses. In addition, we identified the regulatory relationship between DEGs and their transcription factors using in silico and qRT-PCR analysis. Our findings suggest that DEGs and their transcription factors may have a potential role in adipogenesis and/or lipid deposition in liver tissues of two pig breeds.

Molecular Profiling of Clinical Features in Breast Cancer Using Principal Component Analysis (주성분 분석 방법을 이용한 유방암의 임상적 특징과 관련된 유전자 분석)

  • Han, Mi-Ryung;Lee, Seok-Ho;Han, Won-Shik;Kim, Mi-Hyeon;Noh, Dong-Young;Kim, Ju-Han
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2004.11a
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    • pp.29-35
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    • 2004
  • 유방암 환자의 임상정보(clinical features)와 cDNA microarray 기술을 이용하여 얻은 유전자 발현 프로파일은 유방암 예후 인자를 찾는 데에 매우 중요하다. 본 논문에서는 임상정보와 유전자 발현 정보를 접목해서 분석하는 방법으로써 주성분 분석(Principal Component Analysis)을 이용하였다. 이 방법은 다변량 자료의 차원을 줄이는 방법으로써, 대용량 실험 데이터로 인해 발생하는 문제점을 해결하기 위하여 많이 쓰이고 있다. 본 연구에서는 주성분 분석을 이용하여 먼저 한국인 유방암 환자 73명의 cDNA microarray 데이터 차원을 줄이고, 이를 통해 얻어진 주성분(Principal Components)과 임상정보 데이터와의 상관관계를 보았다. One-way ANOVA를 이용한 상관관계 분석 결과의 P-value는 permutation test를 통해 검증하였다. 동일한 방법을 estrogen receptor(ER)(+) 환자 20명과 ER(-) 환자 31명에 적용해본 결과, ER(-) 환자 중에서 재발과 관련된 유전자를 찾을 수 있었다. 주성분 분석을 molecular phenotypic profiles of clinical features에 이용한 결과 발견된 유전자는 유방암의 재발과 관련된 예후 인자로서 의미가 있다.

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Identification of Genes Differentially Expressed in Wild Type and Purkinje Cell Degeneration Mice

  • Xiao, Rui;Park, Youngsook;Dirisala, Vijaya R.;Zhang, Ya-Ping;Um, Sang June;Lee, Hoon Taek;Park, Chankyu
    • Molecules and Cells
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    • v.20 no.2
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    • pp.219-227
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    • 2005
  • Purkinje cell degeneration (pcd) mice are characterized by death of virtually all cerebellar Purkinje cells by postnatal day 30. In this study, we used DNA microarray analysis to investigate differences in gene expression between the brains of wild type and pcd mice on postnatal day 20, before the appearance of clear-cut phenotypic abnormalities. We identified 300 differentially expressed genes, most of which were involved in metabolic and physiological processes. Among the differentially expressed genes were several calcium binding proteins including calbindin-28k, paravalbumin, matrix gamma-carboxyglutamate protein and synaptotagamins 1 and 13, suggesting the involvement of abnormal $Ca^{2+}$ signaling in the pcd phenotype.

1q21.1 microdeletion identified by chromosomal microarray in a newborn with upper airway obstruction

  • Kim, Yoon Hwa;Yang, Ju Seok;Lee, Young Joo;Bae, Mi Hye;Park, Kyung Hee;Lee, Dong Hyung;Shin, Kyung-Hwa;Kim, Seung Chul
    • Journal of Genetic Medicine
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    • v.15 no.1
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    • pp.34-37
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    • 2018
  • A 1q21.1 microdeletion is an extremely rare chromosomal abnormality that results in phenotypic diversity and incomplete penetrance. Patients with a 1q21.1 microdeletion exhibit neurological-psychiatric problems, microcephaly, epilepsy, facial dysmorphism, cataract, and thrombocytopenia absent radius syndrome. We reported a neonate with confirmed intrauterine growth restriction (IUGR), micrognathia, glossoptosis, upper airway obstruction, facial dysmorphism, and eye abnormality at birth as well as developmental delay at the age of 1 year. These clinical manifestations, except for the IUGR and upper airway obstruction, in the neonate indicated a 1q21.1 microdeletion. Here, we report a rare case of a 1q21.1 microdeletion obtained via paternal inheritance in a newborn with upper airway obstruction caused by glossoptosis and tracheal stenosis.

Gene Expression Profiles Related with TCDD-Induced Hepatotoxicity

  • Ryu, Yeon-Mi;Kim, Ki-Nam;Kim, Yu-Ri;Sohn, Sung-Hwa;Seo, Sang-Hui;Lee, Seung-Ho;Kim, Hye-Won;Won, Nam-Hee;Kim, Meyoung-Kon
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.164-171
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    • 2005
  • Toxicological studies have an object of detecting adverse effects of a chemical on an organism based on observed toxicity marker (i.e., serum biochemical markers and chemical-specific gene expression) or phenotypic outcome. To date, most toxicogenomic studies concentrated on hepatic toxicity. cDNA microarray analysis enable discrimination of the responses in animals exposed to different classes of hepatotoxicants. In an effort to further characterize the mechanisms of 2, 3, 7, 8,-Tetrachlorodibenzo-p-dioxin (TCDD or dioxin)-mediated toxicity, comprehensive temporal-responsive microarray analyses were performed on hepatic tissue from Sprague-Dawley rats treated with TCDD. Hepatic gene expression profiles were monitored using custom DNA chip containing 490 cDNA clones related with toxicology. Gene expression analysis identified 26 features which exhibited a significant change. In this study, we observed that the genes related with oxidative stress in rats exposed to Dioxin, such as CYPIIA3 and glutathione S-transferase, were up-regulated at 24hr after exposure. In this study, we carried out to discover novel evidence for previously unknown gene expression patterns related to mechanism of hepatic toxicity in rats exposed to dioxin, and to elucidate the effects of dioxin on the gene expression after exposure to dioxin.

DNA microarray analysis of RNAi plant regulated expression of NtROS2a gene encoding cytosine DNA demethylation (시토신 탈메틸화 관련 NtROS2a 유전자 발현을 제어한 RNAi 식물의 DNA microarray 분석)

  • Choi, Jang Sun;Lee, In Hye;Jung, Yu Jin;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.231-239
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    • 2016
  • To study the transcript levels of epigenetically regulated genes in tobacco, we have developed a transgenic line OX1 overexpressing NtROS2a gene encoding cytosine DNA demethylation and a RNAi plant line RNAi13. It has been reported that salt- and $H_2O_2$-stress tolerance of these transgenic lines are enhanced with various phenotypic characters (Lee et al. 2015). In this paper, we conducted microarray analysis with Agilent Tobacco 4 x 44K oligo chip by using overexpression line OX1, RNAi plant line RNAi 13, and wild type plant WT. Differentially expressed genes (DEGs) related to metabolism, nutrient supply, and various stressed were up-regulated by approximately 1.5- to 80- fold. DEGs related to co-enzymes, metabolism, and methylation functional genes were down-regulated by approximately 0.03- to 0.7- fold. qRT-PCR analysis showed that the transcript levels of several candidate genes in OX1 and RNAi lines were significantly (p < 0.05) higher than those in WT, such as genes encoding KH domain-containing protein, MADS-box protein, and Zinc phosphodiesterase ELAC protein. On the other hand, several genes such as those encoding pentatricopeptide (PPR) repeat-containing protein, histone deacetylase HDAC3 protein, and protein kinase were decreased by approximately 0.4- to 1.0- fold. This study showed that NtROS2a gene encoding DNA glycosylase related to demethylation could regulate adaptive response of tobacco at transcriptional level.

Exposing Zebrafish to Silver Nanoparticles during Caudal Fin Regeneration Disrupts Caudal Fin Growth and p53 Signaling

  • Yeo, Min-Kyeong;Pak, Se-Wha
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.311-317
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    • 2008
  • Zebrafish were exposed to commercial silver nanoparticles (${\sim}$10-20 nm) at 0.4 and 4 ppm during cadual fin regeneration. The silver was in the $Ag^+$ ionic form. Fin regeneration was slow in the group exposed to the lower concentration. The cadual fin, gill, and muscle were assayed after 48 hours and subjected to histological analysis. In all tissues sampled, fish exposed to nanoparticles exhibited infiltration, large mitochondria with empty matrices, and accumulation of nano-sized silver in blood vessels. The results suggested mitochondrial damage and induction of inflammation. Microarray analysis of RNA from young zebrafish (52 hours post-fertilization) that were exposed to nanometer-sized silver particles, showed alteration in expression of the p53 gene pathway related to apoptosis. Gene expression changes in the nanoparticle-treated zebrafish led to phenotypic changes, reflecting increased apoptosis.

Molecular Cytogenetic Characterization of Supernumerary Marker Chromosomes by Chromosomal Microarray (염색체 마이크로어레이를 이용한 표지염색체의 분자세포유전학적 특성)

  • Bae, Mi-Hyun;Yoo, Han-Wook;Lee, Jin-Ok;Hong, Maria;Seo, Eul-Ju
    • Journal of Genetic Medicine
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    • v.8 no.2
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    • pp.119-124
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    • 2011
  • Purpose: Supernumerary marker chromosome (SMC) could be associated with various phenotypic abnormalities based on the chromosomal origin of SMCs. The present study aimed to determine the genomic contents of SMCs using chromosomal microarray and to analyze molecular cytogenetic characterizations and clinical phenotypes in patients with SMCs. Materials and Methods: Among patients with SMCs detected in routine chromosomal analysis, SMCs originating from chromosome 15 were excluded from the present study. CGH-based oligonucleotide chromosomal microarray was performed in 4 patients. Results: The chromosomal origins of SMCs were identified in 3 patients. Case 1 had a SMC of 16.1 Mb in 1q21.1-q23.3. Case 2 showed 21 Mb gain in 19p13.11-q13.12. Case 3 had a 4.5 Mb-sized SMC rearranged from 2 regions of 2.5 Mb in 22q11.1-q11.21 and 2.0 Mb in 22q11.22-q11.23. Conclusion: Case 1 presented a wide range of phenotypic abnormalities including the phenotype of 1q21.1 duplication syndrome. In case 2, Asperger-like symptoms are apparently related to 19p12-q13.11, hearing problems and strabismus to 19p13.11 and other features to 19q13.12. Compared with cat-eye syndrome type I and 22q11.2 microduplication syndrome, anal atresia in case 3 is likely related to 22q11.1-q11.21 while other features are related to 22q11.22-q11.23. Analyzing SMCs using high-resolution chromosomal microarray can help identify specific gene contents and to offer proper genetic counseling by determining genotype-phenotype correlations.

Isolation and Identification of a New Gene Related to Salt Tolerance in Chinese Cabbage (배추에서 신규 염 저항성 관련 유전자 분리 및 검정)

  • Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.748-755
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    • 2013
  • This study was conducted to find a salt tolerance gene in Brassica rapa. In order to meet this objective, we analyzed data from a KBGP-24K oligo chip [BrEMD (Brassica rapa EST and microarray database)] of the B. rapa ssp. pekinensis 'Chiifu' under salt stress (250 mM NaCl). From the B. rapa KBGP-24K microarray chip analysis, 202 salt-responsive unigenes were primarily selected under salt stress. Of these, a gene with unknown function but known full-length sequence was chosen to closely investigate the gene function. The selected gene was named BrSSR (B. rapa salt stress resistance). BrSSR contains a 285 bp open reading frame encoding a putative 94-amino acid protein, and a DUF581 domain. The pSL94 vector was designed to over-express BrSSR, and was used to transform tobacco plants for salt tolerance analysis. T1 transgenic tobacco plants that over-expressed BrSSR were selected by PCR and DNA blot analyses. Quantitative real-time RT PCR revealed that the expression of BrSSR in transgenic tobacco plants increased by approximately 3.8-fold. Similar results were obtained by RNA blot analysis. Phenotypic characteristics analysis showed that transgenic tobacco plants with over-expressed BrSSR were more salt-tolerant than the wild type control under 250 mM NaCl for 5 days. Based on these results, we hypothesized that the over-expression of BrSSR may be closely related to the enhancement of salt tolerance.