• Title/Summary/Keyword: ITS-PCR

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Complete Nucleotide Sequence of KCNE1 in Korean Genome

  • Yeo, Shin-Il;Kim, Su-Won;Kim, Yoon-Nyun;You, Kwan-Hee;Shin, Song-Woo;Kim, Myoung-Hee;Song, Jae-Chan;Yoo, Min
    • Biomedical Science Letters
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    • v.8 no.3
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    • pp.185-188
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    • 2002
  • We have cloned the gene fur long QT syndrome in Korean genome and determined its detailed nucleotide sequence. Blood DNAs were isolated from 68 healthy individuals (including males and females) and the genomic DNAs were amplified by PCR method followed by automatic DNA sequencing. Entire sequence of the coding region for KCNEI was located in exon 3. PCR products were reexamined for the confirmation of KCNE1-specific amplification by nested PCR. KCNE1 mRNA was 436 bp. This corresponded to 129 amino acids. There was no recognizable difference between males and females. This study should contribute to the better understanding of long QT syndrome in Korean population.

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A Current Source using the 12-Pulse Phase-Controlled Rectifier (12-펼스 위상제어 정류기를 사용한 전류원)

  • 송의호;권봉환
    • The Transactions of the Korean Institute of Electrical Engineers
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    • v.39 no.6
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    • pp.545-556
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    • 1990
  • A design method of a current source using 12-pulse phase-controlled rectifier (PCR) is presented. The critical inductance of the 12-pulse PCR is derived and it is shown that the critical inductance can be reduced using a current source. The control circuit of the 12-pulse PCR with an inner fast dynamic loop is proposed to give the frequency synchronism and to reduce the subharmonics due to the unbalance of the transformer of the power line. This circuit is analyzed and its dynamic loop is optimized. The optimal constant PIMF (proportional, integral and measurable variable feedback, and feedforware) controller is also designed using the time-weighted quadratic performance index. It is shown via experimental results that the proposed design method gives high dynamic and static performance of the current source using the 12-pulse PCR.

A Partial Nucleotide Sequence of Chitin Synthase (CHS) Gene from Rice Blast Fungus, Pyricularia oryzae and Its Cloning

  • Hwang, Cher-Won;Park, In-Cheol;Yeh, Wan-Hae;Takagi, Masamchi;Ryu, Jin-Chang
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.157-159
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    • 1997
  • A 340-bp chitin synthase gene(CHS) fragment was cloned from the genomic DNA of Pyricularia oryzae using a PCR process with two primer DNAs corresponding to highly conserved sequences within fungal CHS genes. The entire DNA nucleotide sequences of the cloned DNA fragment were determined and analyzed. The amino acid sequences deduced from the nucleotide sequence of the amplified DNA fragment showed 86% homology to that of the Aspergillus fumigatus CHSE gene (9). Using this PCR-amplified DNA, about 2.3 kb of including the PCR fragment of CHSE gene was cloned from genomic library.

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Rice Transformation by DNA Imbibition and Construction of Plant Vector (DNA imbibition을 이용한 벼의 형질전환과 vector 개발)

  • 유준희;남홍길정구흥
    • KSBB Journal
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    • v.8 no.2
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    • pp.104-109
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    • 1993
  • A vector for plant transformation which had two reporter genes(Gus and Hpt genes) in a single plasmid was constructed. After rice embryos imbibed DNA solution, DNA uptake and gene expression in rice were monitored. Main expression sites of the Gus gene were meristem of root and coleoptiles. There was no difference in Hpt gene expression between a single Hpt vector and the constructed vector in viability of rice in the hygromycin medium after DNA imbibition, The genomic DNA and total RNA extracted from rice transformant survived in the hygromycin medium were subjected to PCR and RT PCR analysis, respectively. As a result, we found the existence of the Hpt gene and its expression in rice.

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Prevalence of Helicobacter species infection in dogs (개의 Helicobacter species 감염 실태)

  • Nam, Hun-woo;Kim, Doo
    • Korean Journal of Veterinary Research
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    • v.40 no.4
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    • pp.747-753
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    • 2000
  • Helicobacter species have commonly been isolated from the gastric mucosa of humans and animals, however have not been known its association with clinical signs. This study was aimed to detect and identify Helicobacter species in the canine stomach by urease test and polymerase chain reaction(PCR). A total of 87 dogs in Kangwon and Kyunggi areas from August, 1998 to June, 1999 were examined. The detection rate of Helicobacter species by urease test for fundal biopsy samples was 83.9%, and positive rate was increased as incubation time was increased. Helicobacter species was detected in the seventy seven dogs(88.5%) of total 87 dogs by PCR. The fifty five strains of the 77 strains of Helicobacter species were identified as H heilmannii and the three strains were identified as H felis by PCR, but the nineteen strains were not identified.

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Detection of Fish Killing Dinoflagellates Cochlodinium polykrikoides and Karlodinium veneficum (Dinophyceae) in the East China Sea by Real-time PCR

  • Park, Tae-Gyu;Kang, Yang-Soon;Park, Young-Tae;Bae, Heon-Meen;Lee, Yoon
    • ALGAE
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    • v.24 no.2
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    • pp.105-110
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    • 2009
  • The rDNAs of figh-killing dinoflagellates Cochlodinium polykrikoides and Karlodinium veneficum were detected from the East China Sea by species-specific real-time PCR probes. Sequence analysesusing the partial ITS sequences from the real-time PCR products showed identical sequences with C. Polykrikoides and K. veneficum, respectively and low expectation values (E-value) of less than 1e-5 suggesting the presence of these organisms in the East Ching Sea shelf water that flows into the Tsushima Strait and the Yellow Sea.

Genetic Diversity of Schistosoma haematobium Eggs Isolated from Human Urine in Sudan

  • Quan, Juan-Hua;Choi, In-Wook;Ismail, Hassan Ahmed Hassan Ahmed;Mohamed, Abdoelohab Saed;Jeong, Hoo-Gn;Lee, Jin-Su;Hong, Sung-Tae;Yong, Tai-Soon;Cha, Guang-Ho;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.271-277
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    • 2015
  • The genetic diversity of Schistosoma haematobium remains largely unstudied in comparison to that of Schistosoma mansoni. To characterize the extent of genetic diversity in S. haematobium among its definitive host (humans), we collected S. haematobium eggs from the urine of 73 infected schoolchildren at 5 primary schools in White Nile State, Sudan, and then performed a randomly amplified polymorphic DNA marker ITS2 by PCR-RFLP analysis. Among 73 S. haematobium egg-positive cases, 13 were selected based on the presence of the S. haematobium satellite markers A4 and B2 in their genomic DNA, and used for RFLP analysis. The 13 samples were subjected to an RFLP analysis of the S. haematobium ITS2 region; however, there was no variation in size among the fragments. Compared to the ITS2 sequences obtained for S. haematobium from Kenya, the nucleotide sequences of the ITS2 regions of S. haematobium from 4 areas in Sudan were consistent with those from Kenya (> 99%). In this study, we demonstrate for the first time that most of the S. haematobium population in Sudan consists of a pan-African S. haematobium genotype; however, we also report the discovery of Kenyan strain inflow into White Nile, Sudan.

Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification

  • Jeong, Joojin;Cho, Sang-Yun;Lee, Wang-Hyu;Lee, Kui-jae;Ju, Ho-Jong
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.219-225
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    • 2015
  • The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.

Development of Species-Specific Primers for Plasmodiophora brassicae, Clubroot Pathogen of Kimchi Cabbage (배추 뿌리혹병균 Plasmodiophora brassicae의 종 특이적 프라이머 개발)

  • Choi, Jin Su;Yang, Seul Gi;Song, Jeong Young;Kim, Hong Gi
    • Research in Plant Disease
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    • v.20 no.1
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    • pp.21-24
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    • 2014
  • Clubroot caused by the obligate biotrophic protist Plasmodiophora brassicae Woronin is one of the most damaging diseases of Brassicaceae family. In this study, we developed species-specific primer sets for rapid and accurate detection of P. brassicae. The primer sets developed amplified a specific fragment only from P. brassicae DNA while they did not amplify a band from 10 other soilborne pathogens or from Kimchi cabbage. In sensitivity test, the species-specific primer set ITS1-1/ITS1-2 could work for approximately 10 spores/ml of genomic DNA showing more sensitivity and accuracy than previous methods. With quantitative real-time PCR test, the primer set detected less spores of P. brassicae than before, confirming that the species-specific primer set could be useful for rapid and accurate detection of P. brassicae.

Development of Nucleic Acid Lateral Flow Immunoassay for Rapid and Accurate Detection of Chikungunya Virus in Indonesia

  • Ajie, Mandala;Pascapurnama, Dyshelly Nurkartika;Prodjosoewojo, Susantina;Kusumawardani, Shinta;Djauhari, Hofiya;Handali, Sukwan;Alisjahbana, Bachti;Chaidir, Lidya
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1716-1721
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    • 2021
  • Chikungunya fever is an arboviral disease caused by the Chikungunya virus (CHIKV). The disease has similar clinical manifestations with other acute febrile illnesses which complicates differential diagnosis in low-resource settings. We aimed to develop a rapid test for CHIKV detection based on the nucleic acid lateral flow immunoassay technology. The system consists of a primer set that recognizes the E1 region of the CHIKV genome and test strips in an enclosed cassette which are used to detect amplicons labeled with FITC/biotin. Amplification of the viral genome was done using open-source PCR, a low-cost open-source thermal cycler. Assay performance was evaluated using a panel of RNA isolated from patients' blood with confirmed CHIKV (n = 8) and dengue virus (n = 20) infection. The open-source PCR-NALFIA platform had a limit of detection of 10 RNA copies/ml. The assay had a sensitivity and specificity of 100% (95% CI: 67.56% - 100%) and 100% (95% CI: 83.89% - 100%), respectively, compared to reference standards of any positive virus culture on C6/36 cell lines and/or qRT-PCR. Further evaluation of its performance using a larger sample size may provide important data to extend its usefulness, especially its utilization in the peripheral healthcare facilities with scarce resources and outbreak situations.