• Title/Summary/Keyword: reverse transcriptase-polymerase chain reaction (RT-PCR)

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자리공 항바이러스 단백질 II 유전자의 형질전환에 의한 연초의 바이러스 저항성 품종 개발 (I)

  • 강신웅;이영기;이기원;박성원;이청호
    • Journal of the Korean Society of Tobacco Science
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    • v.21 no.1
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    • pp.57-63
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    • 1999
  • Pokeweed antiviral protein II (PAP-II) encoding cDNA was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from Phytolacca american a leaf. The PAP-II cDNA fragment of 974bp was subcloned to pBluescript II SK- SmaI site and the inserted PAP-II cDNA fragment was sequenced by dideoxy sequencing method. The number of nucleotides of PAP-II cDNA coding region containing start and stop codon was 933bp. To develop a virus-resistant tobacco plant, PAP-II cDNA fragment was inserted to pKGT101B and the insertion of PAP-II cDNA fragment was confirmed by restriction enzyme analysis and colony PCR.

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Cloning and Expression Analysis of a Grape asr gene, VlASR Containing a Promoter Region. (포도 VIASR 유전자 프로모터의 분리 및 발현 분석)

  • Kihl, Joon-Yeong;Pyee, Jae-Ho
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1157-1165
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    • 2007
  • VvMSA, a grapevine ASR which is highly inducible by sugar and abscisic acid signals was previously shown to be a transcription factor for a hexose transporter gene VvHT1. We isolated a cDNA clone, VlASR which is regulated temporally during the grape berry development by ACP RT-PCR (annealing control primer reverse transcriptase-polymerase chain reaction) and it proved identical to VvMSA. RT-PCR and real-time PCR analyses revealed that the VlASR gene was expressed in berries at fruit set and that its expression increased as berries aged but decreased at the late ripening stage. In order to understand the regulatory mechanism of the asr gene, a genomic fragment was cloned from grapevine. The genomic DNA was 1375 bp long and a sugar box (sucrose box 3 and sucrose responsive element 1) was identified in the 611 bp upstream region of the open reading frame. Analysis of the VlASR promoter::reporter gene fusion demonstrated that this promoter was expressed in transgenic Arabidopsis even without sucrose treatment. This result suggests that the ASR/VvHT1-mediated sugar/ABA signaling, previously reported in grapevine, may not function in Arabidopsis which has no ASR homologue.

Acute viral gastroenteritis in children hospitalized in Iksan, Korea during December 2010-June 2011

  • So, Cheol Whoan;Kim, Dong Sup;Yu, Seung Taek;Cho, Ji-Hyun;Kim, Jong Duck
    • Clinical and Experimental Pediatrics
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    • v.56 no.9
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    • pp.383-388
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    • 2013
  • Purpose: Viral etiology is common in cases of children with acute diarrhea, and antibiotic therapy is usually not required. Therefore, it is important to determine the distribution of common viruses among children hospitalized with acute diarrhea. Methods: We included 186 children who suffered from acute diarrhea and were hospitalized at the Wonkwang University Hospital Pediatric ward from December 1, 2010 to June 30, 2011 in this study. Stool samples were collected and multiplex reverse transcriptase polymerase chain reaction (multiplex RT-PCR) was used to simultaneously determine the viral etiology such as rotavirus, norovirus, astrovirus, or adenovirus. Results: Causative viruses were detected in 72 of the 186 cases (38.7%). The mean age of the virus-positive cases was 1 year and 9 months (range, 1 month to 11 years). Rotavirus was detected in 50/186 (26.9%); norovirus, in 18/186 (9.7%); and astrovirus, in 3/186 cases (1.6%). Adenovirus was not detected in any of the cases. Proportions of norovirus genogroups I and II were 21.1% and 78.9%, respectively. Four of the 51 rotavirus-positive cases (7.8%) had received rotavirus vaccination at least once. The mean duration of diarrhea was 2.8 days (range, 1 to 10 days) and vomiting occurred in 39 of the 72 cases (54.2%). Conclusion: Viral etiology was confirmed in about one-third of the children with acute diarrhea, and the most common viral agent was rotavirus, followed by norovirus.

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
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    • v.2 no.1
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    • pp.109-115
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    • 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.

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Development of Recombinant Saccharomyces cerevisiae Expressing Epoxide Hydrolase for the Preparation of Chiral Phenyl Oxirane (광학활성 Phenyl Oxirane 제조용 유전자 재조합 생촉매 개발)

  • 이수정;이은정;김초희;이지원;김희숙;이은열
    • Journal of Life Science
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    • v.13 no.1
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    • pp.105-109
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    • 2003
  • Epoxide hydrolase (EH) gene from Aspergillus niger #33 was cloned from cDNA library generated by reverse transcriptase-polymerase chain reaction (RT-PCR). The nucleotide sequence analysis revealed that the deduced amino acid sequence was almost similiar to that of A. niger LCP521 previously reported. The cloned EH gene was transformed into Saccharomyces cerevisiae and expressed by addition of galactose. The recombinant S. cerevisiae showed hydrolytic activity toward racemic phenyl oxirane substrate based on chiral GC analysis, and can be used as a potential biocatalyst for the preparation of chiral phenyl oxirane.

Presence of infectious bronchitis virus in Korea before 1986 (1986년 이전 국내 전염성 기관지염 바이러스의 확인)

  • Kwon, Hyuk-joon;Lee, Dong-woo;Ahn, Young-ki;Yoon, Jong-ung;Kim, Sun-joong
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.59-65
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    • 2001
  • To clarify for the presence of infectious bronchitis virus (IBV) in Korea before 1986, in which the virus was first isolated, materials collected from chicken diagnostic consignments between 1980 and 1985 and propagated in chicken embryos or cell cultures were screened by the reverse transcriptase polymerase chain reaction (RT-PCR) targeted to the nucleocapsid gene of the virus. Among 11 samples examined, one sample (IBV-SNU80108) submitted in 1980 showed specific PCR product (281 bp). When the amplified product was sequenced, together with IBV vaccine virus H120 strain, and compared with the data for ten other IBV strains derived from the GeneBank, identities between IBV-SNU80108 and other strains in nucleotide and amino acid sequences ranged 96.3% to 63.7% and 96.4% to 69%, respectively. IBV-SNU80108 was distinct from H120 strain by showing 91.9% and 92.9% identities in the respective sequences. This data suggested that IBV genetically distinctive from other foreign IBV strains might be present before 1986 in Korea.

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Implementation of a Deep Learning-Based Computer-Aided Detection System for the Interpretation of Chest Radiographs in Patients Suspected for COVID-19

  • Eui Jin Hwang;Hyungjin Kim;Soon Ho Yoon;Jin Mo Goo;Chang Min Park
    • Korean Journal of Radiology
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    • v.21 no.10
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    • pp.1150-1160
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    • 2020
  • Objective: To describe the experience of implementing a deep learning-based computer-aided detection (CAD) system for the interpretation of chest X-ray radiographs (CXR) of suspected coronavirus disease (COVID-19) patients and investigate the diagnostic performance of CXR interpretation with CAD assistance. Materials and Methods: In this single-center retrospective study, initial CXR of patients with suspected or confirmed COVID-19 were investigated. A commercialized deep learning-based CAD system that can identify various abnormalities on CXR was implemented for the interpretation of CXR in daily practice. The diagnostic performance of radiologists with CAD assistance were evaluated based on two different reference standards: 1) real-time reverse transcriptase-polymerase chain reaction (rRT-PCR) results for COVID-19 and 2) pulmonary abnormality suggesting pneumonia on chest CT. The turnaround times (TATs) of radiology reports for CXR and rRT-PCR results were also evaluated. Results: Among 332 patients (male:female, 173:159; mean age, 57 years) with available rRT-PCR results, 16 patients (4.8%) were diagnosed with COVID-19. Using CXR, radiologists with CAD assistance identified rRT-PCR positive COVID-19 patients with sensitivity and specificity of 68.8% and 66.7%, respectively. Among 119 patients (male:female, 75:44; mean age, 69 years) with available chest CTs, radiologists assisted by CAD reported pneumonia on CXR with a sensitivity of 81.5% and a specificity of 72.3%. The TATs of CXR reports were significantly shorter than those of rRT-PCR results (median 51 vs. 507 minutes; p < 0.001). Conclusion: Radiologists with CAD assistance could identify patients with rRT-PCR-positive COVID-19 or pneumonia on CXR with a reasonably acceptable performance. In patients suspected with COVID-19, CXR had much faster TATs than rRT-PCRs.

Expression of AmA1 Gene Encoding Storage Protein of Amaranthus in Transgenic Tobacco (형질전환 담배에서 Amaranthus 저장단백질인 AmA1 유전자의 발현)

  • 김태금;김영숙;권태호
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.169-173
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    • 2000
  • A 1,183bp cDNA, AmA1, encoding the seed storage protein of Amaranthus hypochondriacus was isolated by reverse transcriptase-polymerase chain reaction (RT-PCR) and characterized. AmA1 gene was subcloned into plant binary vector under Cauliflower Mosaic Virus (CaMV) 35S promoter and nopaline synthase terminator (3'NOS). The recombinant binary vector was used to transform Nicotiana tabacum using Agrobacterium tumefacien -mediated transformation procedure. Shoots were induced on MS medium with 0.1 mg/L NAA, 1.0 mg/L BA, 100 mg/L kanamycin and 250 mg/L cefotaxime. Transgenic plants were selected on rooting medium based on MS medium containing 200 mg/L kanamycin and 250 mg/L cefotaxime without phytoregulators. The presence of AmA1 gene in the transgenic plants was confirmed by PCR followed by DNA hybridization. The expression of AmA1 gene in the transgenic plant was observed by RT-PCR method.

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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.

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.