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

Search Result 915, Processing Time 0.028 seconds

Effects of Ganoderma lucidum on the IL-1, TNF and IL-12 Gene Expression of Macrophages (영지버섯이 탐식세포의 IL-1, TNF 및 IL-12 유전자 발현에 미치는 영향)

  • 배지현
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.26 no.5
    • /
    • pp.978-982
    • /
    • 1997
  • In order to investigate the immunomodulatory mechanism of Ganoderma lucidum, the effects of protein-bound polysacchride of Ganoderma lucidum on the proliferation and cytokine gene expression of mouse peritoneal macrophages was studied. In the macrophage proliferation assay using the BrdU labeling reagent, the GLA component extracted from Ganoderma lucidum or GLB from the bud of Ganoderma lucidum were added to the medium at the concentration of 0 to 256ug/ml. DNA synthesis of the macrophage was increased at 16ug/ml of GLA and 64ug/ml of GLB, respectively. In the reverse transcription polymerase chain reaction(RT-PCR), the cytokine(TNF, IL-1, and IL-12) gene and $\beta$-actin expression were also analyzed. 20$\mu\textrm{g}$/ml of either GLA or GLB increased TNF and IL-1 expression of the macrophages.

  • PDF

Nucleotide Sequence Analysis of Movement Protein Gene from Tobacco Mosaic Virus Korean Pepper (TMV-KP) Strain (담배 모자이크 바이러스 한국고추계통에서 분리한 이동 단백질 유전자의 염기서열 분석)

  • 이재열;정동수;장무웅;최장경
    • Korean Journal Plant Pathology
    • /
    • v.11 no.1
    • /
    • pp.87-90
    • /
    • 1995
  • Complementary DNA of the movement protein (MP) gene of tobacco mosaic virus Korean pepper strain (TMV-KP) was synthesized from purified TMV-KP RNA by using the reverse transcription and polymerase chain reaction (PCR) system. The synthesized double stranded cDNA was cloned into the plasmid pUC9 and transformed into Escherichia coli JM110. The movement protein gene of TMV-KP of the selected clones was subjected to sequence analysis by Sanger's dideoxy chain termination method. The complete sequence of viral MP gene from TMV-KP strain was 807 nucleotides long. The nucleotide of MP gene from TMV-KP has thirteen and two nucleotide differences from TMV vulgarae (TMV-OM) and Korean (TMV-K) strains, respectively. Thus, the nucleotide sequence of TMV-KP MP gene showed higher homology of 99% with that of TMV-K MP gene.

  • PDF

Genetic Analysis of the Envelope Region of Hepatitis G Vrius isolated from Korean Hepatitis Patients (한국인 간염환자에서 분리한 G형 감염바이러스(HGV)의 외피영역의 유전적 다양성 분석)

  • 김종경;장경립
    • Journal of Life Science
    • /
    • v.8 no.4
    • /
    • pp.360-365
    • /
    • 1998
  • The genetic of a recently described virus, hepatitis G virus(HGV) was investigated. HGV envelope 1 (E1) nucleotide sequences isolated from six Korean hepatitis b virus-positive patients by using a reverse transcription-poly-merase chain reaction procedure, were analysed and compared to the seven previously reported HGV isolates. Sequence homology among the Korean isolates was 88-97% whereas among the isolates from different geographic areas was 80-92%, indicating geographical divergence of HGV. Nucleotide substitutions spread uniformly throughiut the E1 fragment. Furthermore, compared to the prototype HGV sequence, frameshift mutations were observed in most of the Korean isolating that a different translating initiation site for the polyprotein exists in the Korean type HGV.

  • PDF

Selection of Peptides Binding to HCV E2 and Inhibiting Viral Infectivity

  • Hong, Hye-Won;Lee, Seong-Wook;Myung, Hee-Joon
    • Journal of Microbiology and Biotechnology
    • /
    • v.20 no.12
    • /
    • pp.1769-1771
    • /
    • 2010
  • The envelope glycoprotein E2 of hepatitis C virus (HCV) binds to various cell surface receptors for viral infection. We performed biopanning against this protein and selected peptides from phage display peptide libraries. Two short peptides, pep7-1 and pep12-1, were selected and their ability to inhibit the infection process was investigated. When pep7-1 was present, the infectivity of HCV particles in cell culture was notably decreased. This decrease was demonstrated by Western blot analysis, immunofluorescence assay, and reverse transcription PCR assay. However, pep12-1 showed little inhibitory effect on HCV infection.

Molecular Cloning and Identification of a Novel Oxygenase Gene Specifically Induced during the Growth of Rhodococcus sp. Strain T104 on Limonene

  • Park, Ki-Young;Kim, Dockyu;Koh, Sung-Cheol;So, Jae-Seong;Kim, Jong-Sul;Kim, Eungbin
    • Journal of Microbiology
    • /
    • v.42 no.2
    • /
    • pp.160-162
    • /
    • 2004
  • Rhodococcus sp. strain T104 is able to utilize both limonene and biphenyl as growth substrates. Fur-thermore, T104 possesses separate pathways for the degradation of limonene and biphenyl. Previously, we found that a gene(s) involved in limonene degradation was also related to indigo-producing ability. To further corroborate this observation, we have cloned and sequenced a 8,842-bp genomic DNA region with four open reading frames, including one for indole oxygenase, which converts indole to indigo (a blue pigment). The reverse transcription PCR data demonstrated that the identified indole oxygenase gene is specifically induced by limonene, thereby implicating this gene in the degradation of limonene by T104.

Effect of White Ginseng on the Function of Mouse Peritoneal Macrophages and their Gene Expression (백삼성분이 마우스 복강 탐식세포의 기능 및 유전자 발현에 미치는 영향)

  • 배지현
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.26 no.6
    • /
    • pp.1252-1257
    • /
    • 1997
  • In order to investigate the immunomodulatory mechanism of white ginseng, the effects of total saponin of Ginsenoside Rb$_2$component on the phagocytosis and reactive oxygen intermediate(ROI) production of mouse peritoneal macrophages were studied. Both phagocytosis assay nitrobluetetrazolium reduction test showed 20$\mu\textrm{g}$/ml concentration of total saponin significantly increased the activity of phagocytosis and production of ROI. Also cytokine gene expression of the macrophages was analyzed using reverse transcription polymerase chain reaction. In the RT-PCR assay, 20$\mu\textrm{g}$/ml concentration of either total saponin or Ginsenoside Rb$_2$increased IL-1 and TNF expression of the macrophages.

  • PDF

Expression of taurine transporter and taurine uptake in mouse osteoblast cell lines

  • Naomi Ishido;Nakashima, Emi Nakashima;Kang, Yonug-Sook
    • Proceedings of the PSK Conference
    • /
    • 2003.10b
    • /
    • pp.124.2-124.2
    • /
    • 2003
  • Taurine is present in a variety of tissue and exhibits many important physiological functions in the cell. Although it is known that many tissues mediate taurine transport, its functions of taurine transport in bone have not been identified yet. In the present study, we investigated the expression of taurine transporter (TauT) and taurine uptake using mouse stromal ST2 cells and osteoblast-like MC3T3-E1 cells, which is bone related cells. Detection of TauT MRNA expression in these cells were performed by reverse transcription polymerase chain reaction (RT-PCR). (omitted)

  • PDF

Diagnosis Case of Viral Hemorrhagic Septicemia (VHS) in Adult Olive Flounder Paralichthys olivaceus (넙치(Paralichthys olivaceus) 성어의 viral hemorrhagic septicemia (VHS) 진단사례)

  • Kim, In-Woo;Cho, Mi Young;Lee, Han-Na;Han, Hyun Ja;Oh, Yun Kyeong;Lee, Soon Jeong;Jee, Bo Young;Myeong, Jeong-In;Won, Kyoung-Mi
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.45 no.6
    • /
    • pp.666-674
    • /
    • 2012
  • We examined the cause of a disease outbreak in adult olive flounder Paralichthys olivaceus, which occurred at a Korean aquaculture farm in Korea in 2011. The principal signs included an expanded abdomen and congested liver, with persistent mortality (a little over two months). At the beginning of the outbreak, farm administrators misjudged the disease as bacterial in origin, because of the aforementioned signs, persistent mortality, and the detection of bacterial species, including Vibrio spp. and Streptococcus spp. Moreover, the detection of viral hemorrhagic septicemia virus (VHSV) by reverse trasnscription-PCR analysis was complicated by use of the VHS-VN primer set, which has been in general use recently, because it produced weak bands in some samples. Therefore, we recommend the use of at least two different primer sets in the diagnosis of VHSV. Our histopathological findings indicate that necrotizing myocarditis could be considered a pathogenic sign of VHSV infection.

Utilization of qPCR Technology in Water Treatment (수질분석에 사용되는 qPCR기술)

  • Kim, Won Jae;Hwang, Yunjung;Lee, Minhye;Chung, Minsub
    • Applied Chemistry for Engineering
    • /
    • v.33 no.3
    • /
    • pp.235-241
    • /
    • 2022
  • According to the World Water Development Report 2015 released by the United Nations, drinking water is expected to decrease by 40% by 2030. This does not mean that the amount of water decreases, but rather that the water source is contaminated due to environmental pollution. Because microbes are deeply related to water quality, the analysis of microbe is very important for water quality management. While the most common method currently used for microbial analysis is microscopic examination of the shape and feature after cell culture, as the gene analysis technology advances, quantitative polymerase chain reaction (qPCR) can be applied to the microscopic microbiological analysis, and the application method has been studied. Among them, a reverse transcription (RT) step enables the analysis of RNA by RT-PCR. Integrated cell culture (ICC)-qPCR shortens the test time by using it with microbial culture analysis, and viability qPCR can reduce the false positive errors of samples collected from natural water source. Multiplex qPCR for improved throughput, and microfluidic qPCR for analysis with limited amount of sample has been developed In this paper, we introduce the case, principle and development direction of the qPCR method applied to the analysis of microorganisms.