• Title/Summary/Keyword: RT-PCR-hybridization

검색결과 123건 처리시간 0.032초

Estrogen Regulate Neuroprotection and PDI Gene Expression in Ischemic Rat Brain

  • Yu, Seong-Jin;Kim, Do-Rim;Kim, Jee-Yun;Youm, Mi-Young;Lee, Chae-Kwan;Kang, Sung-Goo
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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    • pp.69-69
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    • 2003
  • Neuroprotective strategies have been appeared to be effective in a variety of stroke models. One of the major focuses has been related to the activities of estrogen. $17\beta$-estradiol valerate(EV) has been reported to exert neuroprotective effects when administered before an ischemic insult. The purpose of this study was to determine whether EV can protect against brain injury via estrogen receptor. Chronic and acute pretreatment can reduce the ischemic damage of focal cerebral ischemia in OVX rat, indicating that EV may be a new therapeutic class of drugs to prevent neuronal damage associated with cerebral ischemia. RNAs were extracted from the hippocampus of ovariectomized female rat with or without EV. Differential gene expression profiles were revealed(Bone morphogenetic protein type 1A receptor, Protein disulphide isomerase, cytochrome bc-1 complex core P, thiol-specific antioxidant protein). RT-PCR and in situ hybridization were used to validate the relative expression pattern obtained by the cDNA array. This Study was supported by the Korea Science and Engineering Foundation(KOSEF) through the Biohealth Products Research Center(BPRC), Inje University, Korea

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Expression Profiling of Genes involved in the Control of Pluripotency Using CDNA Microarray

  • Lee, Young-Jin;Hong, Seok-Ho;Nah, Hee-Young;Chae, Jai-Hyung;Jung, Ho-Sun;Kim, Beom-Sue;Kim, Chul-Geun
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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    • pp.18-24
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    • 2001
  • To identify genes implicated in the control of pluripotency as well as characteristics of stem cells, we analyzed expression profiles of genes derived from mouse morulas, blastocysts, embryonic stem cells, mesenchymal stem cells, and uterus tissue cDNA microarray. Comparative analyses of their expression profiles identified putative clones that expressed specifically in specific samples or not in a specific sample. The expression pattern of these candidate clones was analyzed using RT-PCR and non-radioactive in situ hybridization. Functional annotation of these clones on pluripotency and stem cells and molecular mechanisms underlying many facets of mammalian development and differentiation.

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Expression Profiling of Genes involved in the Control of Pluripotency Using cDNA Microarray

  • Lee, Young-Jin;Hong, Seok-Ho;Nah, Hee-Young;Chae, Ji-Hyung;Jung, Ho-Sun;Kim, Beom-Sue;Kim, Chul-Geun
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2001년도 추계학술대회 및 정기총회
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    • pp.12-21
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    • 2001
  • To identify genes implicated in the control of pluripotency as well as characteristics of stem cells, we analyzed expression profiles of genes derived from mouse morulas, blastocysts, embryonic stem cells, mesenchymal stem cells, and uterus tissue using cDNA microarray. Comparative analyses of their expression profiles identified putative clones that expressed specifically in specific samples or not in a specific sample. The expression pattern of these condidate clones was analyzed using RT-PCR and non-radioactive in situ hybridization. Functional annotation of these clones on pluripotency and stem cell plasticity is in ongoing. These studies may further our understanding on the nature of the stem cells and molecular mechanisms underlying many facets of mammalian development and differentiation.

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복제한우 폐조직에서 특이 유전자 발현에 관한 연구 (Studies on the Specific Gene Expression in Lung Tissue of the Cloned Hanwoo)

  • 김상환;정덕원;이호준;황수연;민관식;윤종택
    • Reproductive and Developmental Biology
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    • 제33권1호
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    • pp.19-24
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    • 2009
  • This study was conducted to investigate the specific expression genes in the cloned bovine tissues. Donor cells, cloned tissues were analysed by RAPD-RFLP method. The results were detected three genes (CH-U7B, CH-U7M and CH-U7P) in the cloned fetus. It was found a single copy genes by southern hybridization. Sequence analysis of CH-U7M gene was shown 99% homology to a previously reported EST from a cloned bovine fetus. The putative ORF was encode a protein of hydrophobicity index 0.03. Semi-quantitative RT-PCR by using the CH-LS001 specific primer was remarkably detected in the lung tissue of cloned fetus. Further investigation of these genes may provide one of the key information to explain the early death, abnormal fetus, large off-spring and the low pregnancy rate in the production of cloned bovine.

화학요법제 5-flurouracil의 조혈 및 면역독성 억제를 위한 조성물 개발 연구 (Inhibitive Effect of KH against Toxicity Induced by 5-Flurouracil(FU))

  • 양동식;홍민영;성현제;윤유식
    • 한국한의학연구원논문집
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    • 제8권2호통권9호
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    • pp.109-119
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    • 2002
  • The objective on this study was to investigate effect of KH, which was composed of 9 kinds of oriental herbs tonifing the blood, against toxicity of 5-flurouracil(FU) through animal study. Reduction of WBC and platelet after treating S-FU was significantly recovered by KH. KH also rehabilitated immune gene expression of interleukin-2(IL2) and $Interferon-{\gamma}\;(IFN-{\gamma})$ by RT-PCR. In addition, in situ hybridization of spleen showed that KH treatment increased mRNA expression of IL2. In addition to the immune action, antitumor activity on 5-FU was not affected by KH treatment. In conclusion, our study demonstrated that KH alleviated damage of hematopoiesis system induced by f-FU without toss of antitumor activity.

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Expression of Murine Asb-9 During Mouse Spermatogenesis

  • Lee, Man Ryul;Kim, Soo Kyoung;Kim, Jong Soo;Rhim, Si Youn;Kim, Kye-Seong
    • Molecules and Cells
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    • 제26권6호
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    • pp.621-624
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    • 2008
  • We previously showed that Asb-4 and Asb-17 is uniquely expressed in developing male germ cells. A recent report showed that Asb-9 is specifically expressed in the kidney and testes; however, detailed expression patterns in developing germ cells have not been shown. Northern blot analysis in various tissues demonstrated that mAsb-9 was strongly expressed in the testes. Expression analysis by RT-PCR and Northern blot in developing mouse testes indicates that mAsb-9 is expressed from the fourth week after birth to adulthood, with the highest expression in round spermatids. Expression sites were further localized by in situ hybridization in the testes. Pachytene spermatocytes and spermatids expressed mAsb-9 but spermatogonia and generated spermatozoa did not. This study reveals that mAsb-9 could be a specific marker of active spermatogenesis and would be useful for studies of male germ cell development.

Choristoneura fumiferana Granulovirus pk-1: A Baculoviral Protein Kinase

  • Giannopoulos, Paresa N.;Nassoury, Nasha;Lamontagne, Lucie;Guertin, Claude;Rashidan, Kianoush Khajeh
    • BMB Reports
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    • 제38권4호
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    • pp.457-467
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    • 2005
  • Open reading frame (ORF) 3 on the Choristoneura fumiferana granulovirus (ChfuGV), located in the 11 kb fragment of the BamHI genomic bank encodes a predicted 32-kDa putative kinase protein. Bioinformatics analysis on the predicted amino acid sequence of ChfuGV PK-1 revealed the existence of 11 catalytic subdomains. Sequence analysis within the 5'-untranslated region (5'-UTR) of ChfuGV pk-1 indicates the presence of both putative early and late promoter motifs, indicating that pk-1 may be expressed throughout the infection cycle. Promoter sequence analysis reveals that pk-1 is deprived of a TATA box and appears instead to be regulated by other cis-acting transcriptional regulatory elements. Temporal transcription analysis by RT-PCR confirms the appearance of transcripts detected from 2 h p.i. until 72 h p.i. Northern blot hybridization characterizes pk-1 transcription as a 1.2 kb transcript. Homology comparisons reveal that ChfuGV PK-1 protein is most closely related to Phthorimaea operculalla granulovirus (PoGV) with 80% amino acid identity.

A Toxicogenomic Study to Assess Methylmercury-induced Neurotoxicity

  • Kim, Youn-Jung;Yun, Hye-Jung;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2003년도 추계국제학술대회
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    • pp.177-177
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    • 2003
  • Methylmercury (MeHg) is a well-known neurotoxicant that causes severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, suppressive subtractive hybridization (SSH) was performed to identify differentially expressed genes on human neuroblastoma cell line, SH-SY5Y treated with DMSO and MeHg (6.25 uM) for 6 hr. Differentially expressed cDNA clones were sequenced and were screened by dot blot to eliminate false positive clones. 13 of 35 screened genes were confirmed using real time RT-PCR. These genes include EB1,90-kDa heat-shock protein, chromosome condensation-related SMC-associated protein and brain peptide Al, etc. Analysis of these genes may provide an insight into the neurotoxic effects of MeHg in human neuronal cells and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

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Transcriptional profiles of Rhizobium vitis-inoculated and salicylic acid-treated 'Tamnara' grapevines based on microarray analysis

  • Choi, Youn Jung;Yun, Hae Keun
    • Journal of Plant Biotechnology
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    • 제43권1호
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    • pp.37-48
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    • 2016
  • The transcriptional profiles of 'Tamnara' grapevine (Vitis labruscana L.) to Rhizobium vitis were determined using 12,000 gene oligonucleotide microarray chips constructed with 6,776 unigenes based on the EST sequencing. Among them, 95 clones were up-regulated more than three times and 90 were down-regulated more than 5-times in the R. vitis-inoculated grapevines relative to the control vines. Treatment of salicylic acid showed that 337 clones were upregulated and 52 clones were down regulated in grapevines. Microarray analysis, reverse transcription-polymer chain reaction, and slot blot hybridization analysis revealed that 5, 14, and 64 clones were up-regulated and 10, 12, and 61 clones were down-regulated in wounded, salicylic acid-treated, and R. vitis-inoculated 'Tamnara' grapevine leaves, respectively. The expression patterns of ${\beta}$-1,3-glucanase, proline-rich protein, and lipoxygenase genes of 'Tamnara' moderately resistant to R. vitis were similar to those of resistant 'Concord' and 'Delaware' grapevines. However, chalcone synthase genes in 'Tamnara' grapevines showed similar expression patterns to susceptible grapevines 'Neomuscat' and 'Rizamat'. Further expression studies with various clones for each gene should be conducted to elucidate their roles in resistant responses against pathogens or other stimuli in grapevines. These results could provide better resources for understanding the mechanism of defense responses against crown gall disease and clues for identifying new genes that may play a role in defense against R. vitis in grapevines.

Expression Profiling of Genes involved in the Control of Pluripotency Using cDNA Microarray

  • Lee, Young-Jin;Hong, Seok-Ho;Nah, Hee-Young;Chae, Ji-Hyung;Jung, Ho-Sun;Kim, Beom-Sue;Kim, Chul-Geun
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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    • pp.18-24
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    • 2001
  • To identify genes implicated in the control of pluripotency as well as characteristics of stem cells, we analyzed expression profiles of genes derived from mouse morulas, blastocysts, embryonic stem cells, mesenchymal stem cells, and uterus tissue using cDNA microarray. Comparative analyses of their expression profiles identified putative clones that expressed specifically in specific samples or not in a specific sample. The expression pattern of these candidate clones was analyzed using RT-PCR and non-radioactive in situ hybridization. Functional annotation of these clones on pluripotency and stem cell plasticity is in ongoing. These studies may further our understanding on the nature of the stem cells and molecular mechanisms underlying many facets of mammalian development and differentiation.

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