• 제목/요약/키워드: tissue-specific gene expression

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Functional properties of an alternative, tissue-specific promoter for rice NADPH-dependent dihydroflavonol reductase

  • Kim, Joonki;Lee, Hye-Jung;Tyagi, Wricha;Kovach, Michael;Sweeney, Megan;McCouch, Susan;Cho, Yong-Gu
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.163-163
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    • 2017
  • A deletion analysis of the Oryza sativa dihydroflavonol reductase (DFR) promoter defined a 25 bp region (-386 to -362) sufficient to confer pericarp-specific expression of ${\beta}$ -glucuronidase(GUS) reporter gene in transgenic rice. Site-specific mutagenesis of these conserved sequences and subsequent expression analysis in calli which transiently expressed the mutated promoter::GUS gene showed that both bHLH (-386 to -381) and Myb (-368 to -362) binding sites in the DEL3 (-440 to 70) promoter were necessary for complete expression of the GUS gene including the tissue-specific expression of DFR::GUS gene. The GUS gene was expressed well in the mutated Myb (-368 to -362) binding site, but not as strong as in normal condition, implying that the Myb is also necessary to express GUS gene fully. Also, we found the non-epistatic relation between Rc and DFR. There were no changes of expression patterns GUS under the Rc and rc genotypes. Thus, DFR expression might be independent of the presence of functional Rc gene and suggested that Rc and Rd (DFR) share the same pathway controlling the regulation of flavonoid synthesis but not a direct positive transcriptional regulator of DFR gene.

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Tissue-specific gene expression analysis of silkworm (Bombyx mori) by quantitative real-time RT-PCR

  • Park, Seung-Won;Kang, Seok-Woo;Goo, Tae-Won;Kim, Seong-Ryul;Lee, Gwang-Gill;Paik, Soon-Young
    • BMB Reports
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    • 제43권7호
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    • pp.480-484
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    • 2010
  • The Bombyx mori Microarray Database (BmMDB; http://silkworm.swu.edu.cn/microarray) provides information for tissue-specific gene expression by using the whole-genome oligonucleotide microarray in the silkworm. We analyzed the tissue-specific expression patterns in the silk gland, fat body, and midgut five days of fifth instar larvae during the development of B. mori. To verify the tissue-specific expression, analysis was conducted using quantitative Real-time RT-PCR and the highly expressed endogenous Actin RNA as an intrinsic reference. Finally, we confirmed five genes, (sw15872, sw00692, sw20990, sw05300,and sw2250), out of 18 candidates expressed in two different tissues, which was consistent with the data published by Dr. Xiang's group, thereby supporting the BmMDB. Further studies for promoter regions of candidate genes can be applied in creating transgenic silkworms as biomedical insects for use in producing biomaterials, and to serve as well-characterized models for understanding the mechanism for the genetic regulation of tissue-specific development.

The Brassica rapa Rubber Elongation Factor Promoter Regulates Gene Expression During Seedling Growth in Arabidopsis thaliana and Brassica napus

  • Hong, Joon Ki;Lim, Myung-Ho;Kim, Jin A;Kim, Jung Sun;Lee, Seung Bum;Suh, Eun Jung;Lee, Soo In;Lee, Yeon-Hee
    • Plant Breeding and Biotechnology
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    • 제2권3호
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    • pp.289-300
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    • 2014
  • A tissue-specific and developmentally expressed gene was isolated from Chinese cabbage (Brassica rapa L. ssp. pekinensis), designated BrREF (B. rapa Rubber elongation factor). BrREF transcripts were expressed at high levels in seedlings and at low levels in flower buds and roots. To study the activity of this promoter, the 2.2 kb upstream sequence of BrREF gene was fused to a β-glucuronidase (GUS) reporter gene and was introduced into Arabidopsis thaliana and B. napus by Agrobacterium-mediated transformation. Strong expression of GUS driven by the BrREF promoter was detected in the cotyledons and hypocotyls of transgenic plant seedlings, but GUS expression was weak in roots, excluding the root tips. GUS expression in the cotyledons and hypocotyls decreased dramatically as the seedlings matured and was not detected in the tissues of mature plants. During floral development, GUS expression was observed in immature anthers. These findings suggest that the BrREF promoter can modulate the tissue-specific and developmental expression of gene at the early stages of growth and development.

Tissue- and maturity-dependent expression pattern of androgen receptor mRNA in goldfish, Carassius auratus

  • Choi, Cheol-Young;Kim, Soon-Hag;Kim, Bong-Seok
    • 한국어병학회지
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    • 제16권2호
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    • pp.111-118
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    • 2003
  • Androgen plays an important role in the regulation of gonadotropin production in vertebrates . We have investigated the transcriptional pattern of androgen receptor (AR) in a variety of tissues in maturing male and female goldfish by RT-PCR. Specific primer for AR was designed based on goldfish AR gene from the GenBank (accession number AY090897). AR was shown 10 be maturity- and tissue-dependent gene expression pattern in goldfish. In immature male goldfish, significantly higher transcript level of AR was observed in the pituitary und testis , compared [0 brain and liver. Mature male goldfish showed a similar expression pattern to immature male goldfish. Interestingly. when compare to male goldfish, female goldfish showed AR mRNA expression that was found 10 be weak in pituitary, and very low expression in brain. They could not be found 10 have expression in any other tissues. Taken together. the- transcriptional analysis of AR depending on the tissue, sex. and maturity of a goldfish provides the opportunity for the study of goldfish reproductive physiology ,The results provided for the first time a comparison of the tissue distribution of AR mRNA in sexually maturating male and female goldfish.

Hemocyte-specific Promoter for the Development of Transgenic Silkworm, Bombyx mori

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong-Ryul;Choi, Gwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • 제25권1호
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    • pp.111-114
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    • 2012
  • In previous studies we have shown that a sw17255 gene was expressed in hemocyte-specific tissues of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). It was verified that the sw17255 core promoter region contains elements that regulate the expression of this gene in hemocyte tissue; the selected promoter region spans nucleotides -1 to -2,112 upstream of the start codon. Each of the luciferase reporter gene expression vectors under the control of 4 different kinds of promoter candidates, (-2,112/-1), (-1,640/-1), (-1,169/-1) and (-579/-1), and the control reporter plasmid DNA, were introduced into B. mori larval coelom by direct injection using a syringe. The promoter candidate [E] (-579/-1) showed more than 1.67 fold transcriptional activity compared to control promoter activity. Higher productivity of an expressed gene in the transgenic silkworm by this promoter combination could be achieved in the near future. The foreign recombinant protein could be easily harvested from the blood of the transgenic silkworm.

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.

Genes expression monitoring using cDNA microarray: Protocol and Application

  • Muramatsu Masa-aki
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2000년도 국제심포지움 및 추계학술대회
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    • pp.31-41
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    • 2000
  • The major issue in the post genome sequencing era is determination of gene expression patterns in variety of biological systems. A microarray system is a powerful technology for analyzing the expression profile of thousands of genes at one experiment. In this study, we constructed cDNA microarray which carries 2,304 cDNAS derived from oligo-capped mouse cDNA library. Using this hand-made microarray we determined gene expression in various biological systems. To determine tissue specific genes, we compared Nine genes were highly-expressed in adult mouse brain compared to kidney, liver, and skeletal muscle. Tissue distribution analysis using DNA microarray extracted 9 genes that were predominantly expressed in the brain. A database search showed that five of the 9 genes, MBP, SC1, HiAT3, S100 protein-beta, and SNAP25, were previously known to be expressed at high level in the brain and in the nervous system. One gene was highly sequence similar to rat S-Rex-s/human NSP-C, suggesting that the gene is a mouse homologue. The remaining three genes did not match to known genes in the GenBank/EMBL database, indicating that these are novel genes highly-expressed in the brain. Our DNA microarray was also used to detect differentiation specific genes, hormone dependent genes, and transcription-factor-induced genes. We conclude that DNA microarray is an excellent tool for identifying differentially expressed genes.

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구강편평상피암종에서 DCC 유전자의 역할 (ROLE OF DCC(DELETED IN COLORECTAL CANCER) GENE IN ORAL SQUAMOUS CELL CARCINOMA)

  • 고성규;한세진;김경욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제34권5호
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    • pp.518-524
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    • 2008
  • Chromosome 18q alteration plays a key role in colorectal tumorigenesis, and loss of heterozygosity at 18q is associated with a poor prognosis in colon cancer. DCC(Deleted in Colorectal Cancer) is a putative tumor- suppressor gene at 18q21 that encodes a transmembrane protein with structural similarity to neural cell adhesion molecule that is involved in both epithelial and neuronal cell differentiation. DCC is implicated in regulation of cell growth, survival and proliferation. Thus, tumor progression in squamous cell carcinoma, stomach cancer, colorectal cancer correlates with downregulation of DCC expression. The mechanism for DCC suppression is associated with hypermethylation of the DCC gene promoter region. Hence, the goal of this study is to identify the promoter methylation responsible for the down-regulation of DCC expression in oral squamous cell carcinoma. 12 of tissue specimens for the study are excised and gathered from 12 patients who are diagnosed as SCC in department of OMS, dental hospital, dankook university. To find expression of DCC in each tissue samples, immunohistochemical staining, RT-PCR gene analysis and methylation specific PCR are processed. The results are as follows. 1. In the DCC gene RT-PCR analysis, 5(41.6%) of 12 specimens of oral squamous cell carcinoma did not expressed DCC gene. 2. In the promoter methylation specific PCR analysis, 5(41.6%) of 12 specimens showed promoter methylation of DCC gene. 3. In the immunohistochemical staining of poor differentiated and invasive oral squamous cell carcinoma, loss of DCC expression was observed. These findings suggest that methylation of the DCC gene may play a role in loss of gene expression in invasive oral squamous cell carcinoma.

Association of DNA Methylation Levels with Tissue-specific Expression of Adipogenic and Lipogenic Genes in Longissimus dorsi Muscle of Korean Cattle

  • Baik, M.;Vu, T.T.T.;Piao, M.Y.;Kang, H.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권10호
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    • pp.1493-1498
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    • 2014
  • Epigenetic factors, such as DNA methylation status, may regulate adipogenesis and lipogenesis, thus affecting intramuscular fat (IMF) deposition in longissimus dorsi muscle (LM) of beef cattle. In Korean cattle steers, the LM consists mainly of muscle tissue. However, the LM tissue also contains IMF. We compared the gene expression levels between the IMF and muscle portions of the LM after tissue separation. Real-time polymerase chain reaction analysis showed that the mRNA levels of both adipogenic peroxisome proliferator-activated receptor gamma isoform 1 (PPARG1) and lipogenic fatty acid binding protein 4 (FABP4) were higher (p<0.01) in the IMF than in the muscle portion of the LM. We determined DNA methylation levels of regulatory regions of the PPARG1 and FABP4 genes by pyrosequencing of genomic DNA. DNA methylation levels of two of three CpG sites in the PPARG1 gene promoter region were lower (p<0.05) in the IMF than in the muscle portion of the LM. DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion. Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene. Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.