• Title/Summary/Keyword: Transcriptase

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Presence of Diverse Sugarcane Bacilliform Viruses Infecting Sugarcane in China Revealed by Pairwise Sequence Comparisons and Phylogenetic Analysis

  • Ahmad, Kashif;Sun, Sheng-Ren;Chen, Jun-Lu;Huang, Mei-Ting;Fu, Hua-Ying;Gao, San-Ji
    • The Plant Pathology Journal
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    • v.35 no.1
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    • pp.41-50
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    • 2019
  • Sugarcane bacilliform viruses (SCBV), which belong to the genus Badnavirus, family Caulimoviridae, are an important DNA virus complex that infects sugarcane. To explore the genetic diversity of the sugarcane-infecting badnavirus complex in China, we tested 392 sugarcane leaf samples collected from Fujian, Yunnan, and Hainan provinces for the occurrence of SCBV by polymerase chain reaction (PCR) assays using published primers SCBV-F and SCBV-R that target the reverse transcriptase/ribonuclease H (RT/RNase H) regions of the viral genome. A total of 111 PCR-amplified fragments (726 bp) from 63 SCBV-positive samples were cloned and sequenced. A neighbor-joining phylogenetic tree was constructed based on the SCBV sequences from this study and 34 published sequences representing 18 different phylogroups or genotypes (SCBV-A to -R). All SCBV-tested isolates could be classified into 20 SCBV phylogenetic groups from SCBV-A to -T. Of nine SCBV phylogroups reported in this study, two novel phylogroups, SCBV-S and SCBV-T, that share 90.0-93.2% sequence identity and show 0.07-0.11 genetic distance with each other in the RT/RNase H region, are proposed. SCBV-S had 57.6-92.2% sequence identity and 0.09-0.66 genetic distance, while SCBV-T had 58.4-90.0% sequence identity and 0.11-0.63 genetic distance compared with the published SCBV phylogroups. Additionally, two other Badnavirus species, Sugarcane bacilliform MO virus (SCBMOV) and Sugarcane bacilliform IM virus (SCBIMV), which originally clustered in phylogenetic groups SCBV-E and SCBV-F, respectively, are first reported in China. Our findings will help to understand the level of genetic heterogeneity present in the complex of Badnavirus species that infect sugarcane.

Detection of foot-and-mouth disease virus and coxsakievirus in the soil and leachate of modeled carcass burial site (시험 가축 매몰지 토양 및 침출수 내에서의 구제역 바이러스 검출)

  • Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.35 no.4
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    • pp.255-261
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    • 2012
  • Foot and mouth disease (FMD) is highly infectious disease of cloven-hoofed animals, particularly cattle, sheep, pigs and goats. Last outbreak reported in November, 2010 induced the enormous social and economical impacts. Culling of infected animals, movement control, and vaccination are the major control measures of FMD. The aim of this study was to detection foot-and-mouth disease virus (FMDV) in the soil and leachate from modeling burial for pig carcass as measured by real-time reverse transcriptase polymerase chain reaction (RT-PCR). FMDV and Coxsakievirus B1 (CVB1) were detected in soil by week 16 and Coxsakievirus B1 (CVB1) by weeks 12, respectively. FMDV and CVB1 also detected by weeks 8 in the leachate. Results from this study provides an evidence that FMDV could be inactivated for safe of pig carcasses infected with FMDV within 4 month in the carcass burial site.

Paclitaxel Stimulates Cyclooxygenase-2 Expression via MAP Kinase Pathway in Rabbit Articular Chondrocytes

  • Im, Jeong-Hee;Kim, Song-Ja
    • Biomedical Science Letters
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    • v.15 no.2
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    • pp.141-146
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    • 2009
  • Paclitaxel, an antimicrotubule agent, binds to beta-tubulin in the microtubule and stabilizes the polymer, thereby repressing dynamic instability. Here, we have demonstrated that microtubule cytoskeletal architecture involved in regulation of the COX-2 expression in chondrocyte treated with paclitaxel. Paclitaxel enhanced COX-2 expression and prostaglandin E2 production, as indicated by the Western blot analysis, reverse transcriptase PCR(RT-PCR) and immunofluorescence staining, and $PGE_2$ assay, respectively. In our previous data have shown that paclitaxel treatment stimulated activation of ERK-1/2 and p38 kinase(Im et al., 2009). SB203580, an inhibitor of p38 kinase, blocked the induction of COX-2 expression by paclitaxel. Also PD98059, an inhibitor of ERK-1/2 kinase was blocked the induced COX-2 expression. These results indicate that activation of ERK-1/2 and p38 kinase is required for COX-2 expression induced by paclitaxel in rabbit articular chondrocytes.

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Effects of Silkworm Hemolymph and Cartilage-specific Extracellular Matrices on Chondrocytes and Periosteum-derived Progenitor Cells

  • Shin, Hyun-Chong;Choi, Yong-Soo;Lim, Sang-Min;Lee, Chang-Woo;Kim, Dong-Il
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.364-367
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    • 2006
  • In order to enhance the repair of defects in articular cartilage via cell therapy with autologous chondrocytes, as well as with periosteum-derived progenitor cells (PDPCs), silkworm hemolymph (SH) and a variety of cartilage-specific extracellular matrices (ECMs) including type II collagen, proline, chondroitin 4-sulfate, and chondroitin 6-sulfate were assessed with regard to their efficacy as media supplements. SH, a known anti-apoptotic agent, was found to enhance cell growth, as was shown by the results of a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to the results of reverse transcriptase polymerase chain reaction (RT-PCR) analyses, the cartilage-specific ECMs were found to stimulate the expression of hyaline cartilage-specific genes, most notably type II collagen and Sox9, in monolayer cultures of PDPCs.

광학활성 Styrene Oxide 제조를 위한 고기능성 유전자 재조합 Epoxide Hydrolase 생촉매 개발

  • Lee, Su-Jeong;Lee, Ji-Won;Lee, Eun-Jeong;Kim, Hui-Suk;Lee, Eun-Yeol
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.435-438
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    • 2003
  • Epoxide hydrolase(EH) catalyze the enantioselective hydrolysis of racemic epoxides to corresponding diols. A recombinant Pichia pastoris with EH from Rhodotorula glutinis has been constructed by reverse transcriptase-polymerase chain reaction(RT-PCR). The recombinant biocatalyst enantioselectively hydrolyze (R)-styrene oxide faster than (S)-enantiomer. The catalytic activity of recombinant biocatalyst was 7-fold higher than that of wild-type strain. The recombinant EH biocatalyst can be used for kinetic resolution for the production of enantiopure styrene oxide.

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Random Isolation of Cadmium-induced Gene by Reverse Transcriptase PCR in Schizosaccharomyces pombe

  • Choi, Soon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.4 no.1
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    • pp.20-23
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    • 1994
  • The polymerase chain reaction was used to study the genes inducible under stress from the heavy metal cadmium. Schizosaccharomyces pombe, grown in the presence or absence of sublethal concentration of cadmium, was isolated to purify the total RNAs. The Induced RNA Random Fishing (IRRF) method in which random oligonucleotides were used as primers was applied to the identification of cadmium-induced gene expressions. A PCR-DNA product of 400-bp was cloned and sequenced. Computer analysis showed that this DNA has no homology with any known DNA sequences in GenBank or EMBL databases. The induction of this gene was confirmed by Northern blot analysis of total RNAs isolated from both cadmium-treated and untreated yeast cells.

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Cell- and Stage-Specific Expression of the Murine nm23-M5 Gene during Late Spermatogenesis and Spermiogenesis

  • Hwang Gyu-Chan;Ok Do-Won;Lee Mi-Suk;Kim Jin-Hoe
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.5-5
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    • 2002
  • Nucleoside diphosphate kinases (NDPKs) are conserved through evolution and have been shown to be involved in various biological phenomena. By functional screening in yeast, we identified a new member of the NDPK family, ㎚23-M5, which encodes a 211-amino acid protein with 86% identify to the human homolog, ㎚23-H5. Northern blot analysis reveals that ㎚23-M5 encodes two transcripts of 0.8 and 0.7 kb, which are highly and specifically expressed in adult testis. Reverse transcriptase polymerase chain reaction analysis shows that nm23-M5 first appears in pachytene spermatocytes and increase chain reaction in abundance through subsequent stages. (omitted)

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Effects of 17 β -estradiol, bisphenol A and genistein on the expression of the glutathione peroxidase gene of Philasterides dicentrarchii (Ciliophora: Scuticociliata)

  • Lee, Eun-Hye;Kim, Sung-Mi;Nam, Yoon-Kwon;Kim, Ki-Hong
    • Journal of fish pathology
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    • v.19 no.3
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    • pp.189-195
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    • 2006
  • A subtracted cDNA library of a marine scuticociliate, Philasterides dicentrarchii, in response to 17β-estradiol exposure was constructed using suppression subtractive hybridization (SSH). As a result of SSH, 275 clones were isolated, and among them, only glutathione peroxidase (GPX) gene was isolated as an antioxidative enzyme responding to 17β-estradiol. The semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis revealed that the transcription of GPX gene of P. dicentrarchii was clearly increased by exposure to 17β-estradiol. The GPX transcription was also clearly increased by exposure to xenoestrogens such as bisphenol A (BPA) and genistein.

Expression of Enterotoxin Genes in Staphylococcus aureus Isolates Based on mRNA Analysis

  • Lee, Young-Duck;Moon, Bo-Youn;Park, Jong-Hyun;Chang, Hyo-Ihl;Kim, Wang-June
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.461-467
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    • 2007
  • Staphylococcus aureus strains are important foodborne pathogens that produce various toxins. To evaluate the risk of the enterotoxins, four S. aureus strains from kimbap and two clinical samples were isolated and identified, and their expression of the enterotoxin genes were analyzed using a reverse transcription real-time PCR. Various enterotoxin genes were detected, including sea, seg, seh, sei, sen, seo, and sem, where each isolate contained one or two. When the mRNA detection of the enterotoxin genes was analyzed using a reverse transcriptase PCR, various levels of expression were found depending on the species and enterotoxin gene. Therefore, it is reasonable to suggest that the poisoning risk of S. aureus can be effectively evaluated based on the gene expression at the mRNA level.