• Title/Summary/Keyword: Reaction sequence

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Full-Length cDNA Cloning and Nucleotide Sequence Analysis of Cucumber Mosaic Virus (Strain Kor) RNA2

  • Kwon, Chang-Seob;Park, Kyung-Hee;Chung, Won-Il
    • Journal of Plant Biology
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    • v.39 no.4
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    • pp.265-271
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    • 1996
  • Full-length cDNA for RNA2 of cucumber mosaic virus strian Kor (Kor-CMV) was cloned downstream of synthetic T7 promoter by reverse transcriptase-polymerase chain reaction (RT-PCR). The clone could generate a full-length transcript corresponding to RNA1 in size when synthesized by T7 RNA polymerase. The complete nucleotide sequence has shown that the RNA2 is composed of 3,049 nucleotides and contains one functional open reading frame (ORF) of 2,574 nucleotides encoding 2a protein. The deduced translation product of the 2,574 nucleotides contains GDD motif which is a characteristic of RNA-dependent RNA polymerase (RdRp). The amino acid sequence analysis of the 2a protein has shown that the homology is found in decreasing order with O-CMV (98.8%), Y-CMV (98.7%), Fny-CMV (98.3%), KCMV (94.9%), Ix-CMV (91.9%), and Q-CMV (74.9%). Kor-CMV is suggested to belong to subgroup Ⅰ in the aspect of nucleotide sequence homology of RNA2.

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Molecular cloning and sequence Analysis of the Gene for SecY from Streptomyces coelicolor (Muller) (Streptomyces coelicolor에서 secY 유전자의 클로닝과 염기서열 결정)

  • Kim, Sang-Suk;Hyun, Chang-Gu;Kim, Young-Min;Lee, Joo-Hun;Chung, In-Kwon;Kim, Dae-Myung;Suh, Joo-Won
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.678-686
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    • 1995
  • SecY is a central component of the protein export machinery that mediate the translocation of secretory proteins across the plasma membrane of Escherichia coli. In order to study the mechanism of protein secretion in Streptomyces, we have done cloning and sequencing of the Streptomyces coelicolor secY gene by using polymerase chain reaction method. The nucleotide sequence of the gene for SecY from S. coelicolor showed over 58% identity to that of M. luteus. The deduced amino acid sequences were highly homologous to those of other known SecY polypeptides, all having the potential to form 10 transmembrane segments, and especially second, fifth, and tenth segments were particularly conserved, sharing greater than 75% identity with W. lute s SecY. We propose that the conserved membrane-spanning segments actively participate in protein export. In B. subtilis and E. coli, the secY gene is a part of the spc operon, is preceded by the gene coding for ribosomal protein L15, and is likety coupled transcriptionally and translationally to the upstream L15 gene. In the other hand, secY gene of S. coelicolor and M. luteus have its own promoter region, are coupled translationally with adk gene and pr sented in adk operon.

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Molecular Detection of Phytoplasmas of the 16SrI and 16SrXXXII Groups in Elaeocarpus sylvestris Trees with Decline Disease in Jeju Island, South Korea

  • Geon-Woo, Lee;Sang-Sub, Han
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.149-157
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    • 2023
  • Phytoplasmas were discovered in diseased Elaeocarpus sylvestris trees growing on Jeju Island that showed symptoms of yellowing and darkening in the leaves. Leaf samples from 14 symptomatic plants in Jeju-si and Seogwipo-si were collected and phytoplasma 16S rRNA was successfully amplified by nested polymerase chain reaction using universal primers. The sequence analysis detected two phytoplasmas, which showed 99.5% identity to 'Candidatus Phytoplasma asteris' and 'Ca. P. malaysianum' affiliated to 16SrI and 16SrXXXII groups, respectively. Through polymerase chain reaction-restriction fragment length polymorphism (RFLP) analyses using the AfaI (RsaI) restriction enzyme, the presence of two phytoplasmas strains as well as cases of mixed infection of these strains was detected. In a virtual RFLP analysis with 17 restriction enzymes, the 16S rRNA sequence of the 'Ca. P. asteris' strain was found to match the pattern of the 16SrI-B subgroup. In addition, the phytoplasmas in the mixed-infection cases could be distinguished using specific primer sets. In conclusion, this study confirmed mixed infection of two phytoplasmas in one E. sylvestris plant, and also the presence of two phytoplasmas (of the 16SrI and 16SrXXXII groups) in Jeju Island (Republic of Korea).

Development of PCR Assay for Identification of Buffalo Meat

  • Rajapaksha, W.R.A.K.J.S.;Thilakaratne, I.D.S.I.P.;Chandrasiri, A.D.N.;Niroshan, T.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1046-1048
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    • 2003
  • A polymerase chain reaction (PCR) assay was developed to differentiate buffalo meat from the meat of Ceylon spotted deer (Axis axis ceylonensis), Ceylon sambhur (Cervus unicolor unicolor), cattle (Bovine), goat (Caprine), pig (Porcine), and sheep (Ovine). A set of primers were designed according to the sequence of the mitochondrial cytochrome b gene of bubalus bubalis and by PCR amplification a band of approximately 242 bp band was observed with buffalo DNA. These primers did not cross-react with DNA of other animal species tested in the study under the specified reaction conditions. A band of 649 bp was observed for all animal species tested when DNA was amplified with the universal primers indicating the presence of mitochondrial DNA in the samples. The technique was sensitive enough to identify rotten (10 days post slaughter), dried and cooked buffalo meat. The absence of a cross reaction with human DNA using the buffalo specific primers eliminates possible false positive reactions.

Study on the Treatability of High-Concetration Wastewater by ABBR (ASBR에 의한 고농도폐수의 혐기성처리 연구)

  • 김종찬;김요용;김세진;정일현
    • Journal of environmental and Sanitary engineering
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    • v.10 no.1
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    • pp.98-105
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    • 1995
  • In the treatment of wastewater or sewage plant sludge with high solid concentration, high rate digestion process in which heating and mixing occur at a time is mainly used, and in the case of wastewater containing solid matter below 1000mg/ℓ the recently developed AF or UASB is developed Recently and commonly utilized. But these processes have weakpoints such as clogging of packing media and need of long period of trial run after microorganism granulation. In this point of view, there are active researches on the ASBR( anaerobic sequence batch reaction ) that is capable of treating of organic matter with reactor that has no packing materials and controlling the inflow time, reaction time sedimentation time and outflow time by time control without loss of microorganisms. The objectives of this study are to evaluate the efficiency of ASBR process according to the reaction time, change of treated water quality and gas output rate in the treatment of wheat plant wastewater.

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Temperature control of a batch polymerization reactor using nonlinear predictive control algorithm (비선형 예측제어 알고리즘을 이용한 회분식 중합 반응기의 온도제어)

  • 나상섭;노형준;이현구
    • 제어로봇시스템학회:학술대회논문집
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    • 1996.10b
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    • pp.1000-1003
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    • 1996
  • Nonlinear unified predictive control(UPC) algorithm was applied to the temperature control of a batch polymerization reactor for polymethylmethacrylate(PMMA). Before the polymerization reaction is initiated, the parameters of the process model are determined by the recursive least squares(RLS) method. During the reaction, nonlinearities due to generation of heat of reaction and variation of heat transfer coefficients are predicted through the nonlinear model developed. These nonlinearities are added to the process output from the linear process model. And then, the predicted process output is used to calculate the control output sequence. The performance of nonlinear control algorithm was verified by simulation and compared with that of the linear unified predictive control algorithm. In the experiment of a batch PMMA polymerization, nonlinear unified predictive control was implemented to regulate the temperature of the reactor, and the validity of the nonlinear model was verified through the experimental results. The performance of the nonlinear controller turned out to be superior to that of the linear controller for tracking abrupt changes in setpoint.

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Analysis of Lactobacillus casei and Mutant Strains by Polymerase Chain Reaction (Polymerase Chain Reaction에 의한 Lactobacillus casei 및 돌연변이 균주들의 비교 분석)

  • Nam, Jin-Sik;Lee, Jeong-Jun;Shin, Myeong-Su;Na, Seog-Hwan;Baek, Young-Jin;Yoo, Min
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.577-583
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    • 1994
  • To classify Lactobacillus casei strains on the basis of difference in their chromosomal DNA sequence, we have performed polymerase chain reactions on their chromosomal DNA by using random primers, and followed by analyzing randomly amplified polymorphic DNA fragments. We also developed a mini-preparative method to isolate PCR-grade chromosomal DNA from Lacto- bacillus casei strains within 3 hours. Based on RAPD pattems by polymerase chain reactions with degenerated random primers, 4 Lactobacillus casei strains and 2 mutant strains were successfully discriminated. Results were very sensitive, strain-specific and reproducible. It was also reliable. These results suggest that RAPD may be applied efficiently for the identification of several Lactoba- cillus casei strains.

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Detection of Mycoplasma gallisepticum using Polymerase Chain Reaction(PCR) (PCR 기법을 이용한 Mycoplasma gallisepticum의 검출)

  • Lee, Young-ju;Kim, Ki-seuk;Kim, Jong-wan;Tak, Ryun-bin
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.90-95
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    • 1999
  • A species-specific 760 base pair(bp) BamHI to EcoRI DNA fragment(fMG-2) of lipoprotein gene was isolated from a Mycoplasma gallisepticum(M gallisepticum) genomic library. Based on the DNA sequence data of fMG-2, a pair of 25bp primers was synthesized. When used in the polymerase chain reaction(PCR), 732bp DNA products were amplified from 6 standard strains and 10 field isolates of M gallisepticum, but not from 2 Mycoplasma synoviae and 7 other Mycoplasma species. The lower detection limit was 100fg of the genomic DNA. Identity of the PCR products was confirmed by comparison of patterns of restriction endonuclease analysis with AseI, DraI, EcoRV and SspI.

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Sequential pattern load modeling and warning-system plan in modular falsework

  • Peng, Jui-Lin;Wu, Cheng-Lung;Chan, Siu-Lai
    • Structural Engineering and Mechanics
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    • v.16 no.4
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    • pp.441-468
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    • 2003
  • This paper investigates the structural behavior of modular falsework system under sequential pattern loads. Based on the studies of 25 construction sites, the pattern load sequence modeling is defined as models R (rectangle), L and U. The study focuses on the system critical loads, regions of largest reaction forces, discrepancy between the pattern load and the uniform load, and the warning-system plan. The analysis results show that the critical loads of modular falsework systems with sequential pattern loads are very close to those with the uniform load used in design. The regions of largest reaction forces are smaller than those calculated by the uniform load. However, the regions of largest reaction forces of three models under sequential pattern loads can be considered as the crucial positions of warning-system based on the measured index of loading. The positions of the sensors for the warning-system for these three different models are not identical.

Developing species-specific quantitative real-time polymerase chain reaction primers for detecting Lautropia mirabilis

  • Park, Soon-Nang;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.46 no.3
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    • pp.140-145
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    • 2021
  • This study aimed to develop Lautropia mirabilis-specific quantitative real-time polymerase chain reaction (qPCR) primers based on the sequence of DNA-directed RNA polymerase subunit beta gene. The PrimerSelect program was used in designing of the qPCR primers, RTLam-F4 and RTLam-R3. The specificity of the qPCR primers were performed by conventional PCR with 37 strains of 37 oral bacterial species, including L. mirabilis. The sensitivity of the primers was determined by qPCR with the serial dilution of purified genomic DNA of L. mirabilis KCOM 3484, ranged from 4 ng to 4 fg. The data showed that the qPCR primers could detect only L. mirabilis strains and as little as 40 fg of genome DNA of L. mirabilis KCOM 3484. These results indicate that this qPCR primer pair (RTLam-F4/RTLam-R3) may be useful for species-specific detection of L. mirabilis in epidemiological studies of oral bacterial infectious diseases such as periodontal disease.