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

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Detection of Soybean Mosaic Virus Using RT-PCR

  • Kim, Yul-Ho;Kim, Ok-Sun;Lee, Bong-Choon;Roh, Jae-Hwan;Kim, Myoung-Ki;Im, Dae-Joon;Hur, Il-Bong;Lee, Sang-Chul
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.3
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    • pp.253-255
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    • 1999
  • Reverse transcription and polymerase chain reaction (RT-PCR) assay was used to detect SMV strains. A pair of oligonucleotide primers were designed to include the cylindrical inclusion (CI) coding region between 4,176 to 5,560 nt. Amplification from the total RNA extracted from infected plants with SMV yielded a 1,385 bp DNA fragment. RT-PCR was shown to be $10^3$ times more sensitive than the ELISA assay and it could detect a virus in $10^{-6}$ dilution. Restriction enzyme analysis of RT- PCR products using EcoR I showed that SMV isolates were classified into six groups according to the patterns of restriction fragments.

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Isolation and Characterization of Brain-Derived Neurotrophic Factor Gene from Flounder (Paralichthys olivaceus)

  • LEE JAE HYUNG;CHOI TAE-JIN;NAM SOO WAN;KIM YOUNG TAE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.838-843
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    • 2005
  • Brain-derived neurotrophic factor (BDNF) is a small secretory protein and a member of the nerve growth factor (NGF) gene family. We cloned the flounder BDNF gene from a flounder brain cDNA library. The nucleotide sequence of the cloned gene showed an open reading frame (ORF) consisting of 810 bp, corresponding to 269 amino acid residues. The tissue distribution of flounder BDNF was determined by reverse transcription-polymerase chain reaction (RT-PCR) in brain, embryo, and muscle tissues. To express fBDNF using a eukaryotic expression system, we constructed the vector mpCTV-BDNF containing the fBDNF gene and transformed this vector into Chlorella ellipsoidea. Stable integration of introduced DNA was confirmed by PCR analysis of genomic DNA, and mRNA expression in C. ellipsoidae was confirmed by RT-PCR analysis.

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.

Monitoring of Viral Hemorrhagic Septicemia Virus in Seawater-Reared Rainbow Trout Oncorhynchus mykiss (해수 사육 무지개송어(Oncorhynchus mykiss)의 Viral Hemorrhagic Septicemia Virus (VHSV) 모니터링)

  • Kim, Wi-Sik;Kong, Kyoung-hui;Jeon, Young-Ho;Oh, Myung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.5
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    • pp.621-623
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    • 2017
  • Mariculture of rainbow trout Onchorhynchus mykiss has been initiated in or around olive flounder Paralichthys olivaceus farms, where viral hemorrhagic septicemia virus (VHSV) is often detected in some fish. In the present study, we investigated VHSV infection in seawater-reared rainbow trout because VHSV has never been detected in salmonids in Korea. A total of 104 adult fish were tested for the presence of VHSV by reverse transcription-polymerase chain reaction, followed by virus isolation with the fathead minnow caudal trunk cell line. Cytopathic effects were observed in two samples but the virus was identified as infectious hematopoietic necrosis virus. Thus, VHSV was not isolated from seawater-reared rainbow trout.

Enhanced Expression of Aldosterone Synthase and Adrenome-dullin in Left and Right Ventricular Hypertrophy in Rats

  • Lee, Jong-Un;Lee, Sung-Su;Kim, Sun-Mi;Lee, Won-Jung
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.2
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    • pp.121-125
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    • 2002
  • The pathophysiological implications of aldosterone and adrenomedullin in the cardiac ventricular hypertrophy were examined. Male Sprague-Dawley rats were treated with deoxycorticosterone acetate (DOCA)-salt and monocrotaline (MCT) to selectively elicit left and right ventricular (LV, RV) hypertrophy, respectively. The mRNA expression of aldosterone synthase and adrenomedullin in LV and RV was determined by reverse transcription-polymerase chain reaction. The expression of aldosterone synthase and adrenomedullin was increased in LV, while not altered significantly in RV of DOCA-salt-treated rats. On the contrary, the expression was not significantly altered in LV, but increased in RV of MCT-treated rats. The enhanced expression of aldosterone synthase may be causally related with the development of ventricular hypertrophy, and the increased expression of adrenomedullin may act as a counter-regulatory mechanism.

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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Development of Single-tube Multiplex Immunocapture RT-PCR Assay for Simultaneous Detection of Two Pepper Tobamoviruses

  • Kim, Jae-Hyun;Choi, Gug-Seoun;Kim, Jeong-Soo;Lee, Sin-Ho;Choi, Jang-Kyung;Ryu, Ki-Ryun
    • The Plant Pathology Journal
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    • v.22 no.2
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    • pp.164-167
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    • 2006
  • An immunocapture reverse transcription-polymerase chain reaction (IC/RT-PCR) was developed for simultaneous detection of two pepper-infecting RNA viruses, Pepper mud mottle virus (PMMoV) and Tobacco mild green mosaic virus (TMGMV). The assay could be performed in a single tube for simultaneous and sensitive detection of these tobamoviruses. This detection system revealed thousand-fold increase in detection sensitivity compare to ELISA. This method could save time and reagent cost compare to common RT-PCR which needs several reactions and several procedures of viral RNA extractions for the same number of samples.

Characterization and RT-PCR Detection of Turnip Mosaic Virus Isolated from Chinese Cabbage in Korea (배추에서 분리한 순무 모자이크 바이러스의 특성 및 역전사 중합효소 연쇄반응법(RT-PCR)을 이용한 검정)

  • 박원목;최설란;김수중;최승국;류기현
    • Korean Journal Plant Pathology
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    • v.14 no.3
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    • pp.223-228
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    • 1998
  • Turnip mosaic virus)TuMV-Ca) was isolated from a Chinese cabbage showing severe mosaic and black necrotic spots symptoms in Korea. The virus was identified as a strain of TuMV by its host range test, particle morphology, serology, double stranded RNA analysis. For detection of the virus, reverse transcription and polymerase chain reaction(RT-PCR) was performed with a set of 18-mer TuMV-specific primers to amplify a 876 bp DNA fragment The virus was rapidly detected from total nucleic acids of virus infected tissues as well as native viral RNA of purified virion particles by RT-PCR. Detection limit of the viral RNA by RT-PCR was 10 fg.

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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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Differentiation between Porcine Epidemic Diarrhea Virus and Transmissible Gastroenteritis Virus in Formalin-fixed Paraffin-embedded Tissues by Multiplex RT-nested PCR and Comparison with in situ Hybridization

  • Jung, Kwon-il;Kim, Jung-hyun;Chae, Chan-hee
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2003.10a
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    • pp.27-27
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    • 2003
  • Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) infections are considered difficult to distinguish clinically and histopathologically. Prompt differentiation between PEDV- and TGEV-associated enteritis would greatly facilitate the management of disease in countries where PEDV and TGEV are epizootic. Rapid differential diagnosis and treatment are crucial to reducing mortality and morbidity from PEDV- and TGEV-induced enteritis in piglets. The objective for this study was to develop a protocol to differentiate between PEDV and TGEV directly from formalin-fixed, paraffin-embedded tissue, using a multiplex reverse transcription-nested polymerase chain reaction (RT-nPCR) assay. (omitted)

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