• Title/Summary/Keyword: Differential expression analysis

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Transcript profiling of expressed sequence tags from intramuscular fat, longissimus dorsi muscle and liver in Korean cattle (Hanwoo)

  • Lim, Da-Jeong;Lee, Seung-Hwan;Cho, Yong-Min;Yoon, Du-Hak;Shin, Youn-Hee;Kim, Kyu-Won;Park, Hye-Sun;Kim, Hee-Bal
    • BMB Reports
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    • v.43 no.2
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    • pp.115-121
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    • 2010
  • A large data set of Hanwoo (Korean cattle) ESTs was analyzed to obtain differential gene expression results for the following three libraries: intramuscular fat, longissimus dorsi muscle and liver. To better understand the gene expression profiles, we identified differentially expressed genes (DEGs) via digital gene expression analysis. Hierarchical clustering of genes was performed according to their relative abundance within the six separate groups (Hanwoo fat versus non-Hanwoo fat, Hanwoo muscle versus non-Hanwoo muscle and Hanwoo liver versus non-Hanwoo liver), producing detailed patterns of gene expression. We determined the quantitative traits associated with the highly expressed genes. We also provide the first list of putative regulatory elements associated with differential tissue expression in Hanwoo cattle. In addition, we conducted evolutionary analysis that suggests a subset of genes accelerated in the bovine lineage are strongly correlated with their expression in Hanwoo muscle.

Differential Gene Expression in the Pathogenic Strains of Actinobacillus pleuropneumoniae Serotypes 1 and 3

  • Xie, Fang;Zhang, Mingjun;Li, Shuqing;Du, Chongtao;Sun, Changjiang;Han, Wenyu;Zhou, Liang;Lei, Liancheng
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.789-797
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    • 2010
  • The limited information on differential gene expression in the different serotypes of Actinobacillus pleuropneumoniae has significantly hampered the research on the pathogenic mechanisms of this organism and the development of multivalent vaccines against A. pleuropneumoniae infection. To compare the gene expressions in the A. pleuropneumoniae strains CVCC259 (serotype 1) and CVCC261 (serotype 3), we screened the differentially expressed genes in the two strains by performing representational difference analysis (RDA). Northern blot analyses were used to confirm the results of RDA. We identified 22 differentially expressed genes in the CVCC259 strain and 20 differentially expressed genes in the CVCC261 strain, and these genes were classified into 11 groups: (1) genes encoding APX toxins; (2) genes encoding transferrin-binding protein; (3) genes involved in lipopolysaccharide (LPS) biosynthesis; (4) genes encoding autotransporter adhesin; (5) genes involved in metabolism; (6) genes involved in the ATP-binding cassette (ABC) transporter system; (7) genes encoding molecular chaperones; (8) genes involved in bacterial transcription and nucleic acid metabolism; (9) a gene encoding protease; (10) genes encoding lipoprotein/membrane protein; and (11) genes encoding various hypothetical proteins. This is the first report on the systematic application of RDA for the analysis of differential gene expression in A. pleuropneumoniae serotypes 1 and 3. The determination of these differentially expressed genes will serve as an indicator for future research on the pathogenic mechanisms of A. pleuropneumoniae and the development of a multivalent vaccine against A. pleuropneumoniae infection.

Comparison of Expression Profiles between Trophozoite and Cyst of Acanthamoeba castellanii

  • Moon, Eun-Kyung;Kong, Hyun-Hee
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.313-318
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    • 2012
  • Acanthamoeba is an opportunistic pathogen known to cause granulomatous amoebic encephalitis and amebic keratitis. Acanthamoeba exhibits life cycle consisting of trophozoite and cyst, and the cyst is highly resistant to variable antibiotics and therapeutic agents. To understand the encystation mechanism of Acanthamoeba, the expression profiles of trophozoite and cyst were compared by gene ontology (GO) analysis. Ribosomal proteins and cytoskeletal proteins were highly expressed in trophozoite. In cyst, various protease, and signal transduction - and protein turnover - related proteins were highly expressed. These results correlated with eukaryotic orthologous groups (KOG) assignment and microarray analysis of Acanthamoeba trophozoite and cyst ESTs. The information of differential expression profiles of trophozoite and cyst would provide important clues for research on encystation mechanism of cyst forming protozoa including Acanthamoeba.

Identification of Differentially Expressed Genes in Nickel[li]-Treated Normal Rat Kidney Cells

  • Koh, Jae-Ki;Lee, Sang-Han
    • Environmental Mutagens and Carcinogens
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    • v.24 no.2
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    • pp.85-90
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    • 2004
  • Nickel(II) compounds are carcinogenic metals which induce genotoxicity and oxidative stress through the generation of reactive oxygen species. In search of new molecular pathways toward understanding the molecular mechanism of nickel(II)-induced carcinogensis, we performed mRNA differential display analysis using total RNA extracted from nickel(II) acetate-treated normal rat kidney cells (NRK-52E). Cells were exposed for 3 days to 160 and 240 uM nickel(II) concentrations. cDNAs corresponding to mRNAs for which expression levels were altered by nickel(II) were isolated, sequenced, and followed by a GenBank Blast homology search. Specificity of differential expression of cDNAs was determined by RT-PCR and Western blot analysis. Two of them (SH3BGRL3 and FHIT) were down-regulated and one (metallothionein) was up-regulated by nickel(II) treatment. The expression of these mRNAs were nickel(II) concentration-dependent. The levels of FHIT and metallothionein proteins were also consistent with the results for mRNAs. Overall, although the fundamental questions related to function of these genes in nickel(II)-mediated carcinogenicity are not answered, our study suggests that they can be interesting candidates for studies of molecular mechanisms of nickel(II) carcinogenesis.

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Identification of An Antibacterial Gene by Differential Display from Lipopolysaccharide-Stimulated Dung Beetle, Copris tripartitus

  • Suh, Hwa-Jin;Kim, Yeon-Ju;Bang, Hea-Son;Yun, Eun-Young;Kim, Seong-Ryul;Park, Kwan-Ho;Kang, Bo-Ram;Kim, Ik-Soo;Jeon, Jae-Pil;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.17 no.2
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    • pp.223-228
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    • 2008
  • A novel beetle antimicrobial protein from stimulated Copris tripartitus and the corresponding gene were isolated in parallel through differential display-PCR and expression in Escherichia coli. To find cDNA clones responsible for bacteria resistance, the suppression subtractive hybridization and GeneFishing differentially expressed genes system were employed in the dung beetle, Copris tripartitus immunized with lipopolysaccaride. One cDNA clone from eight subtracted clones was selected through dot blot analysis and confirmed by northern blot analysis. The 516-bp, selected cDNA clone was determined by 5' and 3' rapid amplication of cDNA ends and cloned into the GST fusion expression vector pGEX-4T-1 for expression of the protein. The expressed protein was predicted 14.7 kDa and inhibited the growth of gram-negative bacteria such as Escherichia coli and Pseudomonas aeruginosa. These results implied that the expressed protein is related to immune defense mechanism against microorganism.

cDNA Microarray Analysis of the Differential Gene Expression in the Neuropathic Pain and Electroacupuncture Treatment Models

  • Ko, Je-Sang;Na, Doe-Sun;Lee, Young-Han;Shin, Soon-Young;Kim, Ji-Hoon;Hwang, Byung-Gil;Min, Byung-Il;Park, Dong-Suk
    • BMB Reports
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    • v.35 no.4
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    • pp.420-427
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    • 2002
  • Partial nerve injury is the main cause of neuropathic pain disorders in humans. Acupuncture has long been used to relieve pain. It is known to relieve pain by controlling the activities of the autonomic nervous system. Although the mechanism of neuropathic pain and analgesic effects of electroacupuncture (EA) have been studied in a rat model system, its detailed mechanism at the molecular level remains unclear. To identify genes that might serve as either markers or explain these distinct biological functions, a cDNA microarray analysis was used to compare the expression of 8,400 genes among three sample groups. Messenger RNAs that were pooled from the spinal nerves of 7 normal. 7 neuropathic pain, and 7 EA treatment rat models were compared. Sixty-eight genes were differentially expressed more than 2-fold in the neuropathic rat model when compared to the normal, and restored to the normal expression level after the EA treatment. These genes are involved in a number of biological processes, including the signal transduction, gene expression, and nociceptive pathways. Confirmation of the differential gene expression was performed by a dot-blot analysis. Dot-blotting results showed that the opioid receptor sigma was among those genes. This indicates that opioid-signaling events are involved in neuropathic pain and the analgesic effects of EA. The potential application of these data include the identification and characterization of signaling pathways that are involved in the EA treatment, studies on the role of the opioid receptor in neuropathic pain, and further exploration on the role of selected identified genes in animal models.

Altered Protein Expression in Peach (Prunus persica) Following Fruit Bagging

  • Zhang, Wei;Zhao, Xiaomeng;Shi, Mengya;Yang, Aizhen;Hua, Baoguang;Liu, Yueping
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.32-45
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    • 2016
  • Fruit bagging has been widely practiced in peach cultivation to produce high quality and unblemished fruit. Moreover, fruit bagging has been utilized to study the effect of shading on the quality of fruit. We conducted a proteomic analysis on peach fruit to elucidate the biochemical and physiological events that characterize the effect of bagging treatment. Comparative analysis of 2D electrophoresis (2-DE) gels showed that relative protein levels differed significantly at 125 DAFB (days after full bloom), as well as at 133 DAFB in fruit that had been bagged until 125 DAFB, followed by exposure to sunlight. Most of the proteins with altered expression were identified by MALDI TOF/TOF. Twenty-one proteins with differential expression among the groups were identified at 125 DAFB, while thirty proteins with differential expression among the groups were identified at 133 DAFB. The analysis revealed that expression of proteins involved in photosynthesis, stress responses, and biochemical processes influencing metabolism were altered during bagging treatment, suggesting that regulation of the synthesis of carbohydrates, amino acids, and proteins influenced fruit size, solid/acid ratio, and peel color. This work provides the first characterization of proteomic changes in peach in response to fruit bagging treatment. Identifying and tracking protein changes may allow us to better understand the mechanisms underlying the effects of bagging treatment.

Restoration of the adipogenic gene expression by naringenin and naringin in 3T3-L1 adipocytes

  • Dayarathne, Lakshi A.;Ranaweera, Sachithra S.;Natraj, Premkumar;Rajan, Priyanka;Lee, Young Jae;Han, Chang-Hoon
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.55.1-55.17
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    • 2021
  • Background: Naringenin and its glycoside naringin are well known citrus flavonoids with several therapeutic benefits. Although the anti-adipogenic effects of naringenin and naringin have been reported previously, the detailed mechanism underlying their anti-adipogenesis effects is poorly understood. Objectives: This study examined the anti-adipogenic effects of naringenin and naringin by determining differential gene expression patterns in these flavonoids-treated 3T3-L1 adipocytes. Methods: Lipid accumulation and triglyceride (TG) content were determined by Oil red O staining and TG assay. Glucose uptake was measured using a 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose fluorescent d-glucose analog. The phosphorylation levels of AMP-activated protein kinase (AMPK) and acetyl Co-A carboxylase (ACC) were observed via Western blot analysis. Differential gene expressions in 3T3-L1 adipocytes were evaluated via RNA sequencing analysis. Results: Naringenin and naringin inhibited both lipid accumulation and TG content, increased phosphorylation levels of both AMPK and ACC and decreased the expression level of 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR) in 3T3-L1 adipocytes. RNA sequencing analysis revealed that 32 up-regulated (> 2-fold) and 17 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Scd1, Mogat1, Dgat, Lipin1, Cpt1a, and Lepr, were normalized to the control level in naringenin-treated adipocytes. In addition, 25 up-regulated (> 2-fold) and 25 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Fabp5, Scd1, Srebf1, Hmgcs1, Cpt1c, Lepr, and Lrp1, were normalized to the control level by naringin. Conclusions: The results indicate that naringenin and naringin have anti-adipogenic potentials that are achieved by normalizing the expression levels of lipid metabolism-related genes that were perturbed in differentiated 3T3-L1 cells.

Classification of Genes Based on Age-Related Differential Expression in Breast Cancer

  • Lee, Gunhee;Lee, Minho
    • Genomics & Informatics
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    • v.15 no.4
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    • pp.156-161
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    • 2017
  • Transcriptome analysis has been widely used to make biomarker panels to diagnose cancers. In breast cancer, the age of the patient has been known to be associated with clinical features. As clinical transcriptome data have accumulated significantly, we classified all human genes based on age-specific differential expression between normal and breast cancer cells using public data. We retrieved the values for gene expression levels in breast cancer and matched normal cells from The Cancer Genome Atlas. We divided genes into two classes by paired t test without considering age in the first classification. We carried out a secondary classification of genes for each class into eight groups, based on the patterns of the p-values, which were calculated for each of the three age groups we defined. Through this two-step classification, gene expression was eventually grouped into 16 classes. We showed that this classification method could be applied to establish a more accurate prediction model to diagnose breast cancer by comparing the performance of prediction models with different combinations of genes. We expect that our scheme of classification could be used for other types of cancer data.

Proteomic Analysis of Colonic Mucosal Tissue from Tuberculous and Ulcerative Colitis Patients

  • Kwon, Seong-Chun;Won, Kyung-Jong;Jung, Seoung-Hyo;Lee, Kang-Pa;Lee, Dong-Youb;Park, Eun-Seok;Kim, Bok-Yung;Cheon, Gab-Jin;Han, Koon-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.3
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    • pp.193-198
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    • 2012
  • Changes in the expression profiles of specific proteins leads to serious human diseases, including colitis. The proteomic changes related to colitis and the differential expression between tuberculous (TC) and ulcerative colitis (UC) in colon tissue from colitis patients has not been defined. We therefore performed a proteomic analysis of human TC and UC mucosal tissue. Total protein was obtained from the colon mucosal tissue of normal, TC, and UC patients, and resolved by 2-dimensional electrophoresis (2-DE). The results were analyzed with PDQuest using silver staining. We used matrix-assisted laser desorption ionization time-of-flight/time-of-flight spectrometry (MALDI TOF/TOF) to identify proteins differentially expressed in TC and UC. Of the over 1,000 proteins isolated, three in TC tissue and two in UC tissue displayed altered expression when compared to normal tissue. Moreover, two proteins were differentially expressed in a comparative analysis between TC and UC. These were identified as mutant ${\beta}$-actin, ${\alpha}$-enolase and Charcot-Leyden crystal protein. In particular, the expression of ${\alpha}$-enolase was significantly greater in TC compared with normal tissue, but decreased in comparison to UC, implying that ${\alpha}$-enolase may represent a biomarker for differential diagnosis of TC and UC. This study therefore provides a valuable resource for the molecular and diagnostic analysis of human colitis.