• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.529-533
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• 2001

The ${\beta}$-Amylase cDNA fragment from the oomcete Saprolegnia parasitica was cloned by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from conserved ${\beta}$-amylase sequences. The 5'and 3'regions of the $\beta$-amylase gene were amplified using the rapid amplification of cDNA ends (rACE) system. It consisted of an open reading frame of 1,350 bp for a protein of 450 amino acids. Comparison between the genomic and cDNA sequences revealed that the intron was not present in the coding region. The deduced amino acid sequence of the ${\beta}$-amylase gene had a 97% similarity to the ${\beta}$-amylase of Saprolegnia ferax, followed by 41% similarity to those of Arabidopsis thaliana, Hordeum vulgare, and Zea mays. The ${\beta}$-amylase gene was also expressed in Saccharomyces cerevisiae by placing it under the control of the alcohol dehydrogenase gene (ADC1) promoter.

• Plant Resources
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• 제1권1호
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• pp.13-21
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• 1998

A cDNA Fragment encoding iron storage protrin generated by polymerase chain reaction(PCR) using highly conserved regions of ferritin related genes were used to sereen a red pepper cDNA library. cDNA clone was designated as Fp1. Fp1 clone contatines a 5' nontranslated region of 51dp containing stop conds. Down stream from 5' UTP. an open reading frame of 750bp was observed. followed by a 3' UTR of 272bp. The deduces amino acid sequence of red pepper protein(Fp1) showed 84%, 48% and 36% identity with soybean(SolC). human(HuL H) and horse spleen(HoS-L) ferritin mRNA accumulation in response to iron. Ferritin mRNA accumulation was transient and particularly abundant in leaves. reaching a maxmum at 12h. The level of ferritin mRNA in roots was affected to a lesser extent than in leaves.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.263-269
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• 2016

Temperate phages have been suggested to carry virulence factors and other lysogenic conversion genes that play important roles in pathogenicity. In this study, phage TEM123 in wild-type Staphylococcus aureus from food sources was analyzed with respect to its morphology, genome sequence, and antibiotic resistance conversion ability. Phage TEM123 from a mitomycin C-induced lysate of S. aureus was isolated from foods. Morphological analysis under a transmission electron microscope revealed that it belonged to the family Siphoviridae. The genome of phage TEM123 consisted of a double-stranded DNA of 43,786 bp with a G+C content of 34.06%. A bioinformatics analysis of the phage genome identified 43 putative open reading frames (ORFs). ORF1 encoded a protein that was nearly identical to the metallo-β-lactamase enzymes that degrade β-lactam antibiotics. After transduction to S. aureus with phage TEM123, the metallo-β-lactamase gene was confirmed in the transductant by PCR and sequencing analyses. In a β-lactam antibiotic susceptibility test, the transductant was more highly resistant to β-lactam antibiotics than S. aureus S133. Phage TEM123 might play a role in the transfer of β-lactam antibiotic resistance determinants in S. aureus. Therefore, we suggest that the prophage of S. aureus with its exotoxin is a risk factor for food safety in the food chain through lateral gene transfer.

• Journal of Microbiology and Biotechnology
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• 제21권3호
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• pp.277-283
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• 2011

Glyoxalase I catalyzes the conversion of methylglyoxal to S-D-lactoylglutathione in the presence of glutathione. The structural gene of glyoxalase I (GLO1) was cloned from an osmotolerant yeast, Candida magnoliae, which produces a functional sweetener, erythritol, from sucrose. DNA sequence analysis revealed that the uninterrupted open reading frame (ORF) of C. magnoliae GLO1 (CmGLO1) spans 945 bp, corresponding to 315 amino acid residues, and shares 45.2% amino acid sequence identity to Saccharomyces cerevisiae Glo1. The cloned ORF in a multicopy constitutive expression plasmid complemented the glo1 mutation of S. cerevisiae, confirming that it encodes Glo1 in C. magnoliae. The responses of CmGLO1 to environmental stresses were different from those of S. cerevisiae, which only responds to osmotic stress. An enzyme activity assay and reverse transcription polymerase chain reaction revealed that the expression of CmGLO1 is induced by stress inducers such as methylglyoxal, $H_2O_2$, KCl, and NaCl. The GenBank Accession No. for CmGLO1 is HM000001.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.984-992
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• 2013

The entire nucleotide sequence of the xylose reductase (XR) gene in Candida milleri CBS8195 sourdough yeast was determined by degenerate polymerase chain reaction (PCR) and genome walking. The sequence analysis revealed an open-reading frame of 981 bp that encoded 326 amino acids with a predicted molecular mass of 36.7 kDa. The deduced amino acid sequence of XR of C. milleri was 64.7% homologous to that of Kluyveromyces lactis. The cloned XR gene was expressed in Saccharomyces cerevisiae, and the resulting recombinant S. cerevisiae strain produced xylitol from xylose, indicating that the C. milleri XR introduced into S. cerevisiae is functional. An enzymatic activity assay and semiquantitative reverse transcription-PCR revealed that the expression of CmXR was induced by xylose. The GenBank Accession No. for CmXR is KC599203.

• Genomics & Informatics
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• 제8권2호
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• pp.63-69
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• 2010

Monofunctional biosynthetic peptidoglycan transglycosylase (MBPT) catalyzes the formation of the glycan chain in bacterial cell walls from peptidoglycan subunits: N-acetylglucosamine (NAG) and acetylmuramic acid (NAM). Bifunctional glycosyltransferases such as the penicillin binding protein (PBP) have peptidoglycan glycosyltransferase (PGT) on their C terminal end which links together the peptidoglycan subunits while transpeptidase (TP) on the N terminal end cross-links the peptide moieties on the NAM monosaccharide of the peptide subunits to create the bacterial cell wall. The singular function of MBPT resembles the C terminal end of PBP as it too contains and utilizes a similar PGT domain. In this article we analyzed the infectious and non infectious protein sequences of MBPT from 31 different strains of bacteria using a variety of bioinformatic tools. Motif analysis, dot-plot comparison, and phylogenetic analysis identified a number of significant differences between infectious and non-infectious protein sequences. In this paper we have made an attempt to explain, analyze and discuss these differences from an evolutionary perspective. The results of our sequence analysis may open the door for utilizing MBPT as a new target to fight a variety of infectious bacteria.

• Bulletin of the Korean Chemical Society
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• 제18권6호
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• pp.653-656
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• 1997

The new unsymmetric $N_6$ ligand 1-amino-13-(2-pyridyl)-3,6,9,12-tetraazatridecane (aptatd) containing one pyridyl group has been synthesized and characterized by EA, IR, and NMR. Its proton association constants $(log K_H^n)$ and stability constants $(log K_{ML})$ for Co(Ⅱ), Ni(Ⅱ), Cu(Ⅱ), and Zn(Ⅱ) ions were determined at 298.1 K and ionic strength 0.100 mol $dm^{-3}$ (KNO₃) in aqueous solution by potentiometry: log $K_H^1$=8.80, log $K_H^2$=8.49, log $K_H^3$=6.84, log $K_H^4$=4.17, log $K_H^5$=3.47; log $K_{ML}(Co^{2+})$=18.00, log $K_{ML}(Ni^{2+})$=21.31, log $K_{ML}(Cu^{2+})$=23.62, log $K_{ML}(Zn^{2+})$=15.60. The X-ray structure of its nickel(Ⅱ) complex [Ni(aptatd)]$(ClO_4)_2$ are reported: orthorhombic space group Pbca, a=15.715(1) Å, b=14.280(2) Å, c=19.443(2) Å, V=4363.4 (9) ų with Z=8. The geometry around nickel is a distorted octahedron with the pyridine nitrogen atom being cis to the nitrogen atom of the terminal primary amine.

• Journal of Plant Biology
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• 제39권4호
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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.

• Journal of Trauma and Injury
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• 제34권3호
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• pp.212-217
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• 2021

Various medical scenarios have arisen with the prolonged coronavirus disease 2019 (COVID-19) pandemic. In particular, the increasing number of asymptomatic COVID-19 patients has prompted reports of emergency surgical experiences with these patients at regional trauma centers. In this report, we describe an example. A 25-year-old male was admitted to the emergency room after a traffic accident. The patient presented with stuporous mentality, and his vital signs were in the normal range. Lacerations were observed in the left eyebrow area and preauricular area, with hemotympanum in the right ear. Brain computed tomography showed a contusional hemorrhage in the right frontal area and an epidural hematoma in the right temporal area with a compound, comminuted fracture and depressed skull bone. Surgical treatment was planned, and the patient was intubated to prepare for surgery. A blood transfusion was prepared, and a central venous catheter was secured. The initial COVID-19 test administered upon presentation to the emergency room had a positive result, and a confirmatory polymerase chain reaction (PCR) test was administered. The PCR test confirmed a positive result. Emergency surgical treatment was performed because the patient's consciousness gradually deteriorated. The risk of infection was high due to the open and unclean wounds in the skull and brain. We prepared and divided the COVID-19 surgical team, including the patient's transportation team, anesthesia team, and surgical preparation team, for successful surgery without any transmission or morbidity. The patient recovered consciousness after the operation, received close monitoring, and did not show any deterioration due to COVID-19.

• 한국미생물·생명공학회지
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• 제47권1호
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• pp.139-147
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• 2019

Glutamate decarboxylase catalyzes the conversion of glutamate to gamma-aminobutyric acid (GABA), contributing to pH homeostasis through proton consumption. The reaction is the first step toward the GABA shunt. To date, the enzymes involved in the glutamate metabolism of Photobacterium damselae subsp. piscicida have not been elucidated. In this study, an open reading frame of P. damselae subsp. piscicida, showing homology to the glutamate decarboxylase or putative pyridoxal-dependent aspartate 1-decarboxylase genes, was isolated and cloned into an expression vector to produce the recombinant enzyme. Preliminary gas chromatography-mass spectrometry characterization of the purified recombinant enzyme revealed that it catalyzed not only the decarboxylation of glutamate but also the transamination of GABA. This enzyme of P. damselae subsp. piscicida could be bifunctional, combining decarboxylase and transaminase activities in a single polypeptide chain.