• Title/Summary/Keyword: reverse-transcription polymerase chain reaction

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Identification of Nuclear Receptors by RT-PCR in F9 Cells Induced by Ginsenosides

  • Youl-Nam Lee;Shi
    • Journal of Ginseng Research
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    • v.21 no.3
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    • pp.147-152
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    • 1997
  • Ginsenosides $Rh_1$ and $Rh_2$ Induced the differentiation of F9 teratocarcinoma stem cells. These agents are structurally similar to the steroid hormones, therefore, we speculated that the steroid receptor (s) or novel nuclear receptor (s) could be involved in the differentiation process induces by them. Based on this speculation, we tried to alone new nuclear receptors with reverse transcription-polymerase chain reaction (RT-PCR) method by isolating RNA from F9 teratocarcinoma cells induced by ginsenosides. By using RT-PCR with degenerated primers from highly conserved DNA binding domain of nuclear receptors, we identified several nuclear receptors. In northern blot analysis we found that these clones are transcriptionally regulated by ginsenoside Rhl or Rh2 treatment. Further characterizations of these clones are needed to identify the mechanism of gene expression, which has an important role in the differentiation of F9 cells induced by ginsenosides.

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First Report on Poinsettia mosaic virus in Korea

  • Chung, B.N.;Lee, E.K.;Jeong, M.I.;Kim, H.R.
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.220-223
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    • 2004
  • Most plants of commercial poinsettia cultivars grown from cuttings develop mosaic and chlorotic dot symptoms on leaves. Reverse transcription-polymerase chain reaction (RT-PCR) test showed that they were infected with Poinsettia mosaic virus (PnMV). In a survey of commercially grown poinsettias conducted in Korea, PnMV was detected in ten of ten poinsettia cultivars sampled and in 100% of 178 samples tested. The virus has isometric particles and about 29 nm in diameter. Crystalline virus particles were observed in cytoplasm of cells of diseased plants by transmission electron microscopy. Nucleotide sequence of coat protein gene of PnMV- Kl showed 97.3% homology with that of a German isolate. This is the first report on PnMV in Korea.

Cloning and Expression of a Serine Proteinase Gene Fragment from Acanthamoeba culbertsoni

  • Park, Ki-Won;Kim, Tong-Soo;Na, Byoung-Kuk;Song, Chul-Yong
    • BMB Reports
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    • v.31 no.3
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    • pp.303-306
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    • 1998
  • Serine proteinase cDNA fragment from protozoan parasite Acanthamoeba culbertsoni was amplified by the reverse transcription-polymerase chain reaction (RTPCR) using degenerate oligonucleotide primers derived from conserved serine proteinase sequences. The amplified DNA fragment was subcloned and sequenced. The sequence analysis and alignment showed significant sequence similarity to other eukaryotic serine proteinases and conservation of the His, Asp, and Ser residues that form the catalytic triad. The cDNA fragment was cloned into the pGEMEX-1 expression vector and expressed in Escherichia coli. A resulting fusion protein of 56 kDa had proteolytic activity. The fusion protein reacted with sera of mice immunized with purified serine proteinase of A. culbertsoni in Western blot. Immune recognition of the fusion protein by mouse antisera suggested that the fusion protein may be valuable as a diagnostic reagent.

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Cloning of various bioreactive genes from cartilage tissues of Scyliorhinus torazame (두툽상어 연골 조직에서 생리 활성 유전자들의 cDNA 클로닝)

  • 김지태;김명순;장은령;김영진;김규원
    • Journal of Life Science
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    • v.10 no.5
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    • pp.533-541
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    • 2000
  • Compared to mammal including human, many bioreactive genes that regulate various biological events has not been cloned and characterized yet in fishes, especially shark, Scyliorhinus torazame. In orther to isolate genes that regulate physiological processes in cartilaginors fishes, we performed reverse transcription-polymerase chain reaction (RT-PCR) using the RNA of cartilage tissues of Scyliofhinus torazame. The cloned partial genes were 86%, 80%, 73%, 84%, 75%, 79% identical to $\alpha$- actin, 90-kDa heat-shock protein, methyle-neterahydrofolate dehydrogenase-methenyltertrahydrofolate cyclohudrolase-formyltetrahydrofolate synthetase, ubiquitin, glutamine synthetase and connective tissue growth factor genes of human, respectively. They also have similar nucleotide sequence homologues with those of another species. These partial bioreactive genes elucidated in this study may support to studies of phylogenetic analysis based on evolutionary relationships between shark and other species.

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Identification of Differentially Regulated Genes in Bovine Blastocysts using an Annealing Control Primer System

  • Park, Sae-Young;Hwang, Kyu-Chan;Cui, Xiang-Shun;Shin, Mi-Ra;Kim, Eun-Young;Lee, Won-Don;Kim, Nam-Hyung;Park, Sepill;Lim, Jin-Ho
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.229-229
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    • 2004
  • The identification of embryo-specific genes would provide insights into early embryonic development. However, the current methods employed to identify the genes that are expressed at a specific developmental stage are labor intensive and suffer from high rates of false positives. Here we employed a new and accurate reverse transcription-polymerase chain reaction (RT-PCR) technology that involves annealing control primers (ACPs) to identify the genes that are specifically or prominently expressed in bovine early blastocysts and hatched blastocysts produced in vitro. (omitted)

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Chrysanthemum stunt viroid in Dendranthema grandiflorum

  • Chung, Bong-Nam;Park, Gug-Seoun;Kim, Hyun-Ran;Kim, Jeong-Soo
    • The Plant Pathology Journal
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    • v.17 no.4
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    • pp.194-200
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    • 2001
  • Chrysanthemum stunt viroid (CSVd) ws identified in chrysanthemum cv. Chunkwang showing symptoms of stunt with leaf distortion (K1) and stunt with chlorosis of leaves (K2) collected from the main cultivation area of Masan, Kyongnam province in Korea. The specific RNAs related with the diseased chrysanthemums were detected. Full-length 354 bp CSVd cDNAs were amplified from infected tissue by reverse transcription and polymerase chain reaction using a pair of primers specific for CSVd sequence. The amplified cDNA products were analyzed by agarose gel electrophoresis and the specific cDNAs were cloned. Nucleotide sequences of the two CSVd isolates K1 and K2 varied. Phylogenetic analysis of the nucleotide sequences of CSVd isolates indicated that K1 was closely related with J2 and Am 2 isolates. K1 and K2 were transmitted by grafting to Dendranthema grandiflorum cv. Mistletoe, Gynura aurantiaca, and Lycopersicon esculentum cv. Rutgers. This is the first report of CSVd in D. grandiflorum in Korea.

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Development of a PCR Diagnostic System for Iris yellow spot tospovirus in Quarantine

  • Shin, Yong-Gil;Rho, Jae-Young
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.440-444
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    • 2014
  • Iris yellow spot virus (IYSV) is a plant pathogenic virus which has been reported to continuously occur in onion bulbs, allium field crops, seed crops, lisianthus, and irises. In South Korea, IYSV is a "controlled" virus that has not been reported, and inspection is performed when crops of the genus Iris are imported into South Korea. In this study, reverse-transcription polymerase chain reaction (RT-PCR) and nested PCR inspection methods, which can detect IYSV, from imported crops of the genus Iris at quarantine sites, were developed. In addition, a modified positive plasmid, which can be used as a positive control during inspection, was developed. This modified plasmid can facilitate a more accurate inspection by enabling the examination of a laboratory contamination in an inspection system. The inspection methods that were developed in this study are expected to contribute, through the prompt and accurate inspection of IYSV at quarantine sites to the plant quarantine in South Korea.

Identification of Differentially Expressed Genes by Proto-oncogene Protein DEK using Annealing Control Primers

  • Kim, Dong-Wook;Lee, Jae-Hwi;Seo, Sang-Beom
    • Biomolecules & Therapeutics
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    • v.16 no.3
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    • pp.184-189
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    • 2008
  • The proto-oncogene protein DEK has been implicated in various human disease including cancer. We have shown that DEK induces caspase-dependent apoptosis in Drosophila by regulating histone acetylation. Reverse transcription-polymerase chain reaction (RT-PCR) method based on annealing control primers was used to screen and identify differentially expressed genes (DEGs) in DEK overexpressed HeLa cells. Among the genes identified, clusterin and fibrillarin have major role in apoptosis pathway regulation. TFIIIC and RPS24 are implicated in HAT mediated transcriptional initiation and cololectal cancer, respectively. To further analyze DEK's role in apoptosis, multiplex PCR was performed. Caspase-3, -7, and -10 and proapoptotic gene bid were newly identified as possible target genes regulated by DEK expression.

Analysis and Detection of Coast Protein Gene of Barley Yellow Mosaic Virus and Barley Mield Mosaic Virus by RT-PCR (RP-PCR을 이용한 보리누른모자이크바이러스 (BaYMV)와 보리마일드모자이크바이러스(BaMMV)의 외피단백질 유전자 검정 및 해석)

  • 이귀재
    • Korean Journal Plant Pathology
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    • v.14 no.4
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    • pp.314-318
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    • 1998
  • Using the reverse transcription polymerase chain reaction (RT-PCR), a rapid and sensitive assay method for the detection and identification of barley yellow mosaic virus (BaYMV) and barley mild mosaic virus (BaMMV) was adapted. Two units of primers from each virus were selected and used for the determination of two different viruses. PCR fragments of BaYMV (ca. 0.9kb) and BaMMV (ca. 0.8kb) were obtained from the designed method for the assay of BaYMV and BaMMV coat protein. PT-PCR fragments were cloned using vector pT7 Blue and the sequences of the selected clones were analyzed. coat protein of BaYMV and that of BaMMV consisted of 297 amino acids (891 nucleotides) and 251 amino acids (753 nucleotides), respectively. The snalysis of coat protein genes from these two viruses showed that 45.6% of nucleotides sequence ad 34.9% of amino acid in BaYMV were homologous to those in BaMMV.

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Monitoring of viruses (IHHNV, TSV, IMNV, YHV, and CMNV) in cultured whiteleg shrimp (Litopenaeus vannamei) between 2018 and 2019 (2018-2019년 양식산 흰다리새우의 바이러스 (IHHNV, TSV, IMNV, YHV, CMNV) 모니터링)

  • Kokkattunivarthil, Shyam;Kim, Wi-Sik
    • Journal of fish pathology
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    • v.33 no.1
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    • pp.71-75
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    • 2020
  • A survey was conducted to investigate viral infections in 184 whiteleg shrimp (Litopenaeus vannamei) collected from nine farms and one wholesale fish vendor during 2018 and 2019. Gill and abdominal muscle of shrimp were tested for the presence of five viruses, viz. infectious hypodermal and haematopoietic necrosis virus, taura syndrome virus, infectious myonecrosis virus, yellow head virus genotype 1, and covert mortality nodavirus by reverse transcription-polymerase chain reaction (RT-PCR) and PCR. These viruses were not detected in any of 184 samples, screened under the study.