• Title/Summary/Keyword: mRNA differential display

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Molecular Analysis of the Border Cell Differentiation in Root Cap of Pisum sativum L. (완두(Pisum sativum L.) 근관의 생장과 관련된 표피세포의 분화와 유전자 발현)

  • 우호영;장매희
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.169-173
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    • 1995
  • Border cells are differentiated cells which originate from meristematic cells in The root cap. Experimentally border cells can be released from the root cap by a physical treatment, for example dipping the root tip in the waters After 20-25 hours of release, the new border cell layer forms in the root cap. During the border cell differentiation, new gene expressions were observed in the root cap of pea which was determined by mRNA differential display These new gene expressions may be involved in the border cell differentiation Border cells had unique gene expressions which were determined by mRNA differential display, This suggests that border cells are differentiated cells which are different from the other tissues (ie., leaves, stems, roots or root caps).

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Isolation and Identification of Genes Regulated by Iron Using mRNA Differential Display (mRNA differential display를 이용한 철에 의해 조절되는 유전자들의 분리 및 동정)

  • Lee, Jung-Lim;Park, Jong-Hwan;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.42 no.2
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    • pp.123-127
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    • 1999
  • Iron is an essential nutrient but potentially toxic element in human. To identify the effects of iron on the gene expression of mammalian cell, we have isolated several genes that are regulated by iron using the RNA differential display method. RNAs were isolated from HeLa cells treated with iron supplement or iron chelator. A total of 24 genes were isolated and of these, four genes were identified by DNA sequencing and northern blot.

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Identification of Differentially Expressed Genes in the Longissimus Dorsi Muscle Tissue between Duroc and Erhualian Pigs by mRNA Differential Display

  • Pan, P.W.;Zhao, S.H.;Yu, M.;Liu, B.;Xiong, T.A.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1066-1070
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    • 2003
  • In order to identify differentially expressed mRNAs (which represent possible candidates for significant phenotypic variances of muscle growth, meat quality between introduced European and Chinese indigenous pigs) in the longissimus dorsi muscle tissue between adult Duroc and Erhualian pigs, mRNA differential display was performed. Five 3' anchor primers in combination with 20 different 5' arbitrary primers (100 primer sets) were used and nearly 5,000 cDNA bands were examined, among which 10 differential display cDNAs were obtained, cloned and sequenced. Six of the 10 cDNAs showed similarity to identified genes from GenBank and the other 4 had no matches in GenBank. Differential expression was tested by Northern blot hybridization and could be confirmed for 2 cDNAs. The method used in this study provides a useful molecular tool to investigate genetic variation that occurs at the transcriptional level between different breeds.

Differential Display Analysis of Gene Expression Induced under DCA Treatment in Rat Liver

  • Choi, Soon-Yong;Park, Ock-Jin
    • BMB Reports
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    • v.29 no.3
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    • pp.272-275
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    • 1996
  • The expression of genes induced by Dichloroacetate (DCA) treatment was analyzed by mRNA differential display. Purified total RNAs from rat liver treated with saline or DCA (100 mg/100 g b.w.) were reverse transcribed by using a set of oligonucleotide primers. The PCR products were resolved on a denaturing sequencing gel. PCR band representing mRNA expressed specifically in DCA-treated liver was excised and reamplified by PCR. A 120-bp c-DNA clone named IC1 was isolated and the DNA sequence of IC1 was analyzed. IC1 revealed 50% homology with 3' end of a mouse fibroblast growth factor mRNA This result indicates that DCA induces the expression of a gene which has a 50% homology with a Mouse fibroblast growth factor, and expression of this gene might be involved in non genotoxic process caused by DCA.

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Cloning and Characterization of Liver cDNAs That Are Differentially Expressed between Chicken Hybrids and Their Parents

  • Sun, Dong-Xiao;Wang, Dong;Yu, Ying;Zhang, Yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1684-1690
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    • 2005
  • Using mRNA differential display technique, we investigated differential gene expression in hybrids relative to their parents in a diallel cross involving four chicken breeds in order to provide an insight into the molecular basis of heterosis in chicken. The results indicated that there was extensive differential gene expression between chicken F1 hybrids and their parents which was classified into four kinds of patterns as following: (1) bands only detected in hybrid F1; (2) bands only absent in hybrid F1; (3) bands only detected in parent P1 or P2; (4) bands absent in parent P1 or P2. Forty-two differentially expressed cDNAs were cloned and sequenced, and their expression patterns were confirmed by Reverse-Northern dot blot. Sequence analysis and database searches revealed that genes showed differential expression between hybrid and parents were regulatory and functional genes involved in metabolism, mRNA splicing, transcriptional regulation, cell cycles and protein modification. These results indicated that hybridization between two parents can cause changes in expression of a variety of genes. In conclusion, that the altered pattern of gene expression in hybrids may be responsible for heterosis in chickens.

Increased mRNA Related Ovarian Maturation during Induction of Maturational Competence in Red Seabream, Pagrus major (참돔, Pagrus major의 성숙능력 유도시 증가된 난성숙 관련 mRNA)

  • Choi, Cheol-Young;Chang, Young-Jin;Takashima, Fumio
    • Development and Reproduction
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    • v.4 no.1
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    • pp.125-131
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    • 2000
  • This study has used differential display-PCR, to amplify genes transcribed during the ovarian maturation induced by human chorionic gonadotropin (hCG). The cDNA expressed at the times of acquisition of oocyte maturational competence in red seabream (Pagrus major) following treatment with hCG was amplified and cloned. A full-length of cDNA for p. major was isolated using differential display-PCR and 5'RACE. This cDNA clone contained 2,662 nucleotides including the open reading frame that encoded 434 amino acids. Homology analyses, using the GenBank and EMBL general database searches, indicated that the nucleotides sequence of the cDNA does not have high homology with any other genes. This cDNA was judged to be a gene, which induction of maturational competence coincides with increase of mRNA related ovarian maturation. Consensus sequences which were consistent with protein kinase C phosphorylation sites and casein kinase II phosphorylation sites were identified. in vitro, the transcription level of mRNA related ovarian maturation increased between 9hr and 24hr following treatment of ovarian follicles with hCG. It was also increased after GtH-II (300 ng/ml) stimulation. Furthermore, in vivo, mRNA related ovarian maturation was rarely expressed prior to the acquisition of oocyte maturational competence, but was strongly expressed after the acquisition of oocyte maturational competence, suggesting that the hCG induction of maturational competence is brought about by the de novo synthesis of the mRNA related ovarian maturation in p. major.

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Differential Display of mRNA in the Preimplantation Mouse Embryos by Reverse Transcriptase Polymerase Chain Reaction (역전사 연쇄중합반응에 의한 착상전 생쥐난자에서의 상이한 mRNA의 발현조사에 의한 새로운 유전자의 크로닝법)

  • 김진회;박흠대;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.3
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    • pp.199-206
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    • 1994
  • We present here a new PCR-based cloning technique that allows the different PCR products during mouse embryogenesis. Recently, mRNA differential display described by Liang & Pardee (Science 257, 1992) and re-confirmed by Zimermann & Schultz (PNAS 91,1994). This method will detect the appropriate changes in the temporal patterns of expression or in the transition from maternal control to zygotic control as well as the functional difference of embryo with polyspermy or monospermy, the difference of expression between successfully hatched blastocyst and blastocyst failed to hatching, response to agents, and cell cycle regulation. By this methods, we have cloned an eDNA, which showed mouse 2 cell specific expression. Genomic DNA digested with EcoRI showed approximately 15 kb and then showed higher expression in fetal liver rather than adult liver. Furthermore, this gene is likely to have 2 mRNA by alternative splicing.

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Approach for Cloning and Characterization of Blue/White Flower Color Specific cDNA Clones from Two Commelina Species

  • Lee Gunho;Yeon Mooshik;Hur Yoonkang
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.45-50
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    • 2005
  • To clone blue and white flower color specific genes, mRNA differential display was carried out with two different Commelina species, C. communis Linne for blue color and C. coreana Leveille for. leucantha Nakai for white color. Fifty two and 100 cDNA clones specific for blue or white flower color, respectively, were ranging from 200 to 700 bp in size. From the reverse northern blot analysis, 12 and 7 positive clones were selected for blue and white flower, respectively. These clones appear to be novel cDNAs for these Commelina plants, but not color specific. This finding was supported by the northern blot analysis. However, two clones, B18 and B19, derived from blue flowered Commelina were highly expressed than in the white Commelina species, implying that further study will be valuable. The results indicated that both mRNA display experiment and dot blot analysis may not sensitive enough to clone color-determining gene from the plant, leading to explore more advanced method, like high-density colony array study (HDCA).

Molecular Cloning of Differentially Expressed Genes in First Trap Leaf of Dionaea muscipula by Fluorescent Differential Display (형광 Differential Display법에 의한 파리지옥풀 포충잎트랩 특이발현 유전자 탐색)

  • Kang, Kwon-Kyoo;Lee, Keun-Hyang;Park, Jin-Heui;Hong, Kyong-Ei
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.307-313
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    • 2003
  • Fluorescent differential display (FDD) is a method for identifying differentially expressed genes in eukaryotic cells. The mRNA FDD technology works by systematic amplification of the 3' terminal regions of mRNAs. This method involve the reverse transcription using anchored primers designed to bind 5'boundary of the poly A tails, followed by polymerase chain reaction (PCR) amplification with additional upstream primers of arbitrary sequences. The amplified cDNA subpopulations are separated by denaturing polyacrylamide electrophoresis. To identify the genes involved in the development of first trap leaf, we applied a FDD method using mRNAs from leaf base, first trap leaf and flower tissue, respectively. We screened several genes that expressed specifically in first trap leaf. Nucleotide sequence analysis of these genes revealed that these were protease inhibitor (PI), myo-inositol-1-phosphate synthase and lipocalin-type prostaglandin D synthase. Northern blot analysis showed that these genes were expressed specifically in first trap leaf (in vivo and in vitro). FDD could prove to be useful for simultaneous scanning of transcripts from multiple cDNA samples and faster selection of differentially expressed transcripts of interest.