• Title/Summary/Keyword: reverse-transcription-PCR

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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.

Occurrence and Detection of Rice black-streaked dwarf virus in Korea

  • Lee, Bong-Choon;Hong, Yeon-Kyu;Hong, Sung-Jun;Park, Sung-Tae;Lee, Key-Woon
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.172-173
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    • 2005
  • Until now, occurrence of Rice black-streaked dwarf virus (RBSDV) is observed in Gyeongsang provinces, southeastern part of Korea. However, recently, the occurrence of RBSDV is increasing and spreading in Jeonra provinces including Gochang-gun, southwestern part of Korea. RBSDV infected plants showed typical symptoms including stunted, deformed leaves with white waxy or black-streaked swelling along the veins. We extracted viral genomic dsRNA from infected leaves and analyzed dsRNA pattern by polyacrylamide gel electrophoresis. Ten genomic segments with similar sized dsRNAs were observed. We also detected RBSDV by reverse transcription (RT)-PCR using specific primers for S10 from genomic dsRNA and observed amplified DNA fragment specific for RBSDV S10.

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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cDNA Cloning of Farnesoic Acid-Induced Genes in Candida albicans by Differential Display Analysis

  • CHUNG SOON-CHUN;LEE JI-YOON;OH KI-BONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1146-1151
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    • 2005
  • The yeast Candida albicans has a distinguishing feature, dimorphism, which is the ability to switch between two morphological forms: a budding yeast form and a multicellular invasive filamentous form. This ability has been postulated to contribute to the virulence of this organism. Previously, we reported that the yeast-to-hypha transition in this organism is suppressed by farnesoic acid, a morphogenic autoregulatory substance that accumulates in the medium as the cells proliferate. In this study, using a differential display reverse transcription polymerase chain reaction (DDRT-PCR) technique, we have identified several genes induced in C. albicans by farnesoic acid treatment. These observations indicate that farnesoic acid can alter the expressivity of multiple genes, including the DNA replication machinery and cell-cycle-control proteins.

Functional Characterization of the Major Surface Protein of Treponema maltophilum in Human Gingival Fibroblasts

  • Lee, Sung-Hoon;Choi, Bong-Kyu
    • International Journal of Oral Biology
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    • v.30 no.1
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    • pp.31-37
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    • 2005
  • Treponema maltophilum, a Group IV oral spirochete, is associated with periodontitis and endodontic infections. In this study we analyzed the functional role of the major surface protein of this organism (MspA) in human gingival fibroblasts (HGFs). The full-length gene encoding MspA was cloned and expressed in Escherichia coli by using the expression vector pQE-30. The recombinant protein (rMspA) was purified by affinity chromatography with nickel-nitrilotriacetic acid agarose and possible contamination of E. coli endotoxin in rMspA was removed by using polymyxin B-agarose. rMspA significantly induced the expression of pro inflammatory cytokines like IL-6 and IL-8 and intercellular adhesion molecule (ICAM)-1 in HGFs, when analyzed by reverse transcription-PCR, flow cytometry, and enzyme-linked immunosorbent assay. Our results indicate that MspA of T. maltophilum may play an important role in amplifying the local immune response by upregulating the expression of proinflammatory cytokines and ICAM-1.

Effects of Ginseng Saponin on the Cytokine Gene Expression in Human Immune System (인삼 사포닌이 인간면역계 사이토카인 유전자의 발현에 미치는 영향)

  • 박종욱;한인숙
    • Journal of Ginseng Research
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    • v.20 no.1
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    • pp.15-22
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    • 1996
  • In order to investigate the Immunomodulatory effects of ginseng, we have studied the effects of ginseng saponin on the proliferation and cytosine gene expression of human pheripheral blood mononuclear cell (PBMC). In the PBMC proliferation assay, total saponin exhibited proliferation inhibition on the PBMC or phytohemagglutinin(PHA)-stimulated PBMC in a dose-dependent fashion. Immunomodulatory effects of ginseng were further investigated using the cytokine gene expression as the indicators. In the reverse transcription-polymerase chain reaction (RT-PCR) test, interleukin (IL)-1, IL-2, IL-3, IL-4, IL-6, IL-13, granulocyte macrophage-colony stimulating factor, tumor necrosis factor (TNF), migration inhibitory factor and transforming growth factor genes were expressed in the PHA-stimulated PBMC 48 hrs after cell culture. Among expressed cytokines, total saponin could increase the expression of IL-1 and TNF of PBMC without stimulation of PHA. All of ginsenosides, $Rb_1$, $Rb_2$, $Rg_1$, Rc, Re, incresed TNF gene expression. Especially, Rb2 (20 g/ml) showed most prominent effect on TNF gene expression and it also slightly increased IL-1 gene expression of PBMC.

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Effects of swimming on functional recovery and brain-derived neurotrophic factor (BDNF) mRNA expression after sciatic crushed nerve injury in rats

  • Lee Myoung-Hwa;Byun Yong-Hyun;Yoon Bum-Chul;Kim Chang-Ju
    • The Journal of Korean Physical Therapy
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    • v.16 no.2
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    • pp.128-139
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    • 2004
  • 말초신경은 외상이나 질병 등 여러 가지 원인으로 손상되기 쉬우며, 손상의 정도가 심하거나 치료가 지연되는 경우에는 심각한 기능 소실을 초래할 수 있다. 본 연구에서는 수영이 말초신경손상후 운동기능의 회복과 뇌유인성 신경영양인자 (brain-derived neurotrophic factor, BDNF) mRNA의 발현에 미치는 효과를 알아보기 위하여, 흰쥐 좌골신경에 압박 손상을 가하고 수영을 적용한 후 보행궤적분석 (walking track analysis)과 역전사연쇄반응 (reverse transcription-polymerase chain reaction, RT-PCR)을 실시하였다. 그 결과, 좌골신경 압박손상된 쥐는 특징적인 보행패턴을 나타내어 좌골신경기능지수 (sciatic function index, SFI)가 현저히 낮아졌으며, BDNF mRNA의 발현이 증가하였다. 좌골신경 압박 손상후 수영을 한 쥐에서는 SFI가 현저히 향상되었으며, BDNF mRNA의 발현은 억제되었다. 이러한 결과는 말초신경손상후 수영이 BDNF mRNA의 발현을 조절함으로써 기능 회복을 촉진시키는 효과적인 치료방법이 될 수 있음을 제안하고 있다.

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RNAi-mediated reduction of xanthine dehydrogenase results in increased biomass of Arabidopsis seedlings

  • Nakagawa, Ayami;Sakamoto, Atsushi;Takahashi, Misa;Morikawa, Hiromichi
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.356-360
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    • 2005
  • Xanthine dehydrogenase (XDH), a classic enzyme involved in purine catabolism, can catalyze the formation of redox-signaling reactive oxygen and nitrogen species such as superoxide and nitric oxide. We generated transgenic plants of Arabidopsis in which XDH was knocked out by introduction of hairpin RNA-expression vector. Expression analysis by reverse transcription-PCR and in-gel staining of XDH activity revealed that transgenic lines efficiently suppressedXDH expression at the transcriptional level, demonstrating that RNA interference was successfully induced. XDH-suppressed transgenic lines exhibitedincreased biomass production during the growth of seedlings.

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