• Title/Summary/Keyword: transcriptase

Search Result 547, Processing Time 0.026 seconds

Detection of Potato Spindle Tuber Viroid Using RT-PCR Technique (RT-PCR 기법을 이용한 감자 걀쭉 바이로이드 (Potato Spindle Tuber Viroid)의 검정)

  • Joung, Young-Hee;Jeon, Jae-Heung;Choi, Kyung-Hwa;Kim, Hyun-Soon;Joung, Hyouk
    • Korean Journal Plant Pathology
    • /
    • v.13 no.4
    • /
    • pp.205-209
    • /
    • 1997
  • Potato spindle tuber viroid(PSTVd) RNAs were isolated from PSTVd-inoculated potato cv. Superioc and carried out RT-RCR with reverse transcriptase and PSTVd specifie primer pair desigened to amplify the 356 nucleotides of PSTVd genome. As a result, 356 nucleotides PCR products were amplified from PSTVd-inoculated potato cv. Superior. The 356 nucleotides DNA fragment was indeed the PSTVd geneby sequencing analysis. PSTVd could be successfully detected from infected leaf and tuber tissue of potato by using RT-PCR technique. Especially PSTVd was more effectively detected when both downstream and upstream primer were used than only downstream primer was used in RT reaction.

  • PDF

Detection of Resistance Mutation to Lamivudine in HIV-1 Infected Patients (Lamivudine 복용 HIV-1 감염자에게서 내성 돌연변이 검색)

  • Cho, Young-Keol;Sung, Heung-Sup;Lee, Hee-Jung;Kim, Yoo-Kyum;Chi, Hyun-Sook;Cho, Goon-Jae;Kang, Moon-Won
    • The Journal of the Korean Society for Microbiology
    • /
    • v.35 no.2
    • /
    • pp.181-190
    • /
    • 2000
  • To investigate resistance to lamivudine (3TC), we examined the incidence of M184V in 20 HIV-1 patients treated with 3TC for $13.1{\pm}9$ months. Fourteen of 20 patients had been exposed to zidovudine (ZDV) or didanosine (ddI) prior to 3TC therapy. Nested PCR targeting to reverse transcriptase (RT) and direct sequencing were performed for peripheral blood mononuclear cells sampled serially. There were resistance mutations to ZDV in at least 9 patients at baseline, although there was no resistance mutation to 3TC. We could detect M184V in 6 (30%) out of 20 patients. The incidence of M184V increased as the duration of therapy prolongs (13% in samples <12 months; 47% in samples ${\ge}12$ months). The frequency of mutation M184V was higher in patients with previous mutation to ZDV than in patients with wild type. Resistance mutation was not detected in 7 patients. This study shows that resistance to 3TC tends to develop rapidly in patients with baseline mutations or two drugs combination therapy than in those treated simultaneously with triple drugs. This report is the first on resistance to 3TC in Korean AIDS patients.

  • PDF

A Case of Vertical Transmission of Hepatitis C Virus in an Infant of a Mother Who had Hepatitis C during Pregnancy (C형 간염 산모로부터 출생한 영아에서 C형 간염 Virus의 수직 전파 1례)

  • Oh, Sang-Hyun;Kim, Kuk-Hwan;Yang, Eun-Seok;Park, Sang-Kee;Moon, Kyung-Rye
    • Pediatric Gastroenterology, Hepatology & Nutrition
    • /
    • v.2 no.1
    • /
    • pp.109-115
    • /
    • 1999
  • Hepatitis C virus (HCV) has been identified as an important cause of posttransfusion hepatitis, but vertical transmission of chronic infected HCV RNA positive mothers has been documented in some cases. The reports of the risk of perinatal infection have been widely varied in the literature. The authors experienced one case of vertical transmission of HCV in an infant of a mother who had hepatitis C during pregnancy. At admission, HCV RNA (+), Ig G anti HCV (+) and Ig M anti HCV (+) were found in the mother. Also at admission, HCV RNA (+), Ig G anti HCV (+), Ig M anti HCV (+), elevation of liver aminotransferase level and hepatosplenomegaly on ultrasonography were found in the baby on day 31. HCV RNA (-), Ig M anti HCV (-) and normal of liver aminotransferase level were noted on day 250 in the serum of the infant. We used reverse transcriptase polymerase chain reaction (RT-PCR) technique to find a very small amount of HCV RNA in the serum. All the findings suggest vertical transmission of HCV RNA from mother to infant during 3rd trimester of pregnancy.

  • PDF

In vitro Evaluation of Anti-Human Immunodeficiency Virus Activity of Nucleoside Derivatives and Studies on Their Mode of Action (핵산유도체들의 항 Human Immunodeficiency Virus in vitro 약효평가와 작용기전연구)

  • Lee, Chong-Kyo;Kim, Dong-Ki;Kim, Jee-Hyun;Kim, Hae-Soo;Pi, Mi-Kyoung;Park, Jong-Beak;Kim, Baek
    • The Journal of Korean Society of Virology
    • /
    • v.27 no.1
    • /
    • pp.69-75
    • /
    • 1997
  • To evaluate in vitro anti-HIV efficacies of nucleoside derivatives, MT-4 cell line was infected with HIV-1 and HIV-2 respectively and treated with various compounds and the formerly approved drugs such as AZT, d4T, ddC and ddI. CPE method was used to evaluate their antiviral activity. Most dideoxynucleosides, AZT, d4T, ddC and ddI, showed anti-HIV activities against both viruses but no other compounds including anti-herpesvirus drugs did any. Further experiments were carried out to study their inhibitory mechanism of viral adsorption. The results showed no inhibition of syncytium formation due to an interaction between the gp120 expressed in HIV -infected cell surface and CD4 receptor on the uninfected cell surface in the presence of AZT. AZT showed no activity up to $100\;{\mu}g/ml$. Inhibition of reverse transcriptase (RT) in the presence of AZT-triphosphate was tested by using RT expressed in E. coli and purified and its $IC_{50}$ was 4.5 nM.

  • PDF

Molecular biological characterization of the new virus isolated from abnormally swimming salmonid - Reverse transcriptase activity - (이상유영(異常遊泳) 증상(症狀)을 나타내는 연어과 어류(魚類)에서 분리(分離)된 새로운 바이러스의 분자생물학적(分子生物學的) 특성(特性) - 역전사효소 활성 -)

  • Oh, Myung-Joo
    • Journal of fish pathology
    • /
    • v.9 no.2
    • /
    • pp.177-183
    • /
    • 1996
  • A new RNA virus isolated from abnormally swimming behavior has caused mortalities in salmonid fish (Oh et al., 1995 a), A reverse transcriptase (RTase) activity of the virus was determined by using poly r(A) : oligo d(T) as templete : primer. This RTase activity was associated with virus particles of buoyant density of 1.16 g/ml. The virus particles in sucrose fractions were enveloped and were about 85 nm diameter with central electron-dense core. The brain and kidney samples of artificially infected fish showed RTase activity. Virus particle associated proteins about 120, 80, 65, 61, 48, 42, 35, 30, 25, 19, and 15 kDa were observed when examined by polyacrylamide gel electrophoresis analysis. This study showed the presence of a new retrovirus in salmonid fish, which tentatively called RVS (Retrovirus of salmonid).

  • PDF

Inhibition of Telomerase Activity in U937 Human Monocytic Leukemia Cells by Compound K, a Ginseng Saponin Metabolite

  • Kang Kyoung-Ah;Lee Kyoung-Hwa;Chae Sung-Wook;Kim Jeong-Ki;Seo Jung-Yeon;Ham Yong-Ho;Lee Kee-Ho;Kim Bum-Joon;Kim Hee-Sun;Kim Dong-Hyun;Hyun Jin Won
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.11 no.1
    • /
    • pp.7-12
    • /
    • 2006
  • Telomerase activation is detected in most cancerous cells; hence, telomerase is a highly selective target for cancer therapy, which plays an important role in the apoptotic process. We have previously reported that the ginseng saponin metabolite, Compound K (20-O-D-glucopyranosyl-20(S)-protopanaxadiol, IH901), inhibits cell proliferation by inducing apoptosis and cell cycle arrest at the $G_1$ phase. The present study investigated the regulation of telomerase activity in Compound K treated U937 cells. Compound K treatment caused a reduction in telomerase activity and down-regulated the human telomerase reverse transcriptase (hTERT) gene, resulting in the decreased expressions of its protein, and of the c-Myc and Spl proteins (transcription factors of hTERT). These results indicate that the anticancer activity of Compound K could be mediated by inhibition of the telomerase activity.

Induction of S Phase Arrest of the Cell Cycle by Piceatannol is Associated with Inhibition of Telomerase Activity in Human Leukemic U937 Cells (Piceatannol에 의한 인체 혈구암세포의 증식 억제 및 telomerase 활성 저하)

  • Choi, Yung-Hyun
    • Journal of Life Science
    • /
    • v.18 no.1
    • /
    • pp.96-102
    • /
    • 2008
  • Piceatannol is a polyphenol that is found in abundant quantities in grapes and wine. Although recent experimental data revealed the anti-cancer potency of piceatannol, the molecular mechanisms underlying the antileukemic activity have not yet been studied in detail. In the present study, we investigated further possible mechanisms by which piceatannol exerts its anti-proliferative action in cultured human leukemia U937 cells. Exposure of U937 cells to piceatannol resulted in growth inhibition and induction of apoptosis as measured by MTT assay and flow cytometry analysis, which was associated with S phase arrest of the cell cycle. Piceatannol treatment markedly inhibited the activity of telomerase, and the levels of human telomerase reverse transcriptase (hTERT) and telomerase-associated protein-1 (TEP-1), main determinants of the telomerase enzymatic activity, were progressively down-regulated by piceatannol treatment in a dose-dependent fashion. However, the levels of cyclooxygenases (COXs) expression and prostaglandin E2 (PGE2) release were not changed in piceatannol-treated U937 cells. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of piceatannol.

Detection rate and clinical impact of respiratory viruses in children with Kawasaki disease

  • Kim, Ja Hye;Yu, Jeong Jin;Lee, Jina;Kim, Mi-Na;Ko, Hong Ki;Choi, Hyung Soon;Kim, Young-Hwue;Ko, Jae-Kon
    • Clinical and Experimental Pediatrics
    • /
    • v.55 no.12
    • /
    • pp.470-473
    • /
    • 2012
  • Purpose: The purpose of this prospective case-control study was to survey the detection rate of respiratory viruses in children with Kawasaki disease (KD) by using multiplex reverse transcriptase-polymerase chain reaction (RT-PCR), and to investigate the clinical implications of the prevalence of respiratory viruses during the acute phase of KD. Methods: RT-PCR assays were carried out to screen for the presence of respiratory syncytial virus A and B, adenovirus, rhinovirus, parainfluenza viruses 1 to 4, influenza virus A and B, metapneumovirus, bocavirus, coronavirus OC43/229E and NL63, and enterovirus in nasopharyngeal secretions of 55 KD patients and 78 control subjects. Results: Virus detection rates in KD patients and control subjects were 32.7% and 30.8%, respectively (P=0.811). However, there was no significant association between the presence of any of the 15 viruses and the incidence of KD. Comparisons between the 18 patients with positive RT-PCR results and the other 37 KD patients revealed no significant differences in terms of clinical findings (including the prevalence of incomplete presentation of the disease) and coronary artery diameter. Conclusion: A positive RT-PCR for currently epidemic respiratory viruses should not be used as an evidence against the diagnosis of KD. These viruses were not associated with the incomplete presentation of KD and coronary artery dilatation.

Impaired Hippocampal Synaptic Plasticity and Enhanced Excitatory Transmission in a Novel Animal Model of Autism Spectrum Disorders with Telomerase Reverse Transcriptase Overexpression

  • Rhee, Jeehae;Park, Kwanghoon;Kim, Ki Chan;Shin, Chan Young;Chung, ChiHye
    • Molecules and Cells
    • /
    • v.41 no.5
    • /
    • pp.486-494
    • /
    • 2018
  • Recently, we have reported that animals with telomerase reverse transcriptase (TERT) overexpression exhibit reduced social interaction, decreased preference for novel social interaction and poor nest-building behaviors-symptoms that mirror those observed in human autism spectrum disorders (ASD). Overexpression of TERT also alters the excitatory/inhibitory (E/I) ratio in the medial prefrontal cortex. However, the effects of TERT overexpression on hippocampal-dependent learning and synaptic efficacy have not been investigated. In the present study, we employed electrophysiological approaches in combination with behavioral analysis to examine hippocampal function of TERT transgenic (TERT-tg) mice and FVB controls. We found that TERT overexpression results in enhanced hippocampal excitation with no changes in inhibition and significantly impairs long-term synaptic plasticity. Interestingly, the expression levels of phosphorylated CREB and phosphorylated $CaMKII{\alpha}$ were significantly decreased while the expression level of $CaMKII{\alpha}$ was slightly increased in the hippocampus of TERT-overexpressing mice. Our observations highlight the importance of TERT in normal synaptic function and behavior and provide additional information on a novel animal model of ASD associated with TERT overexpression.

Development of Reverse Transcriptase-Polymerase Chain Reaction of fimA Gene to Detect Viable Salmonella in Milk (우유 내 활력있는 Salmonella를 검출하기 위한 fimA 유전자의 역전사중합효소 연쇄반응의 개발)

  • Choi, S.H.;Lee, S.B.
    • Journal of Animal Science and Technology
    • /
    • v.46 no.5
    • /
    • pp.841-848
    • /
    • 2004
  • Rapid detection of viable Salmonella in pasteurized milk is important to protect public health from food poisoning. Reverse transcriptase-polymerase chain reaction(RT-PCR) is recognized as a molecular genetical method to differentiate between live and dead bacteria The RT-PCR in this study was designed to detect specifically viable Salmonella in milk by using the primers whose nucleotide sequences were determined based on fimA gene which encodes the submit of type 1 fimbriae. Treatment of RNA preparation with RNase-free DNase was adequate enough to destroy DNA, which may otherwise be amplified in the RT PCR Seven strains of Salmonella were detected in the RT-PCR but Escherichia coli, Shigella sonnei, Citrobacter freundii, and Klebsiella pneumoniae were not. $10^7/ml$ and $10^6/ml$ of dead Salmonella which were heat-treated in milk were detectable by using the RT-PCR but $10^5{\sim}10/ml$ of the dead bacteria were not. The sensitivity of the RT-PCR in detecting viable Salmonella was 100 cells/ml.