• Title/Summary/Keyword: KB-C2

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Involvement of IS26 Element in the Evolution and Dissemination of $bla_{SHV-2a}$ and $bla_{SHV-12}$ ($bla_{SHV-2a}$$bla_{SHV-12}$ 항균제 내성 유전자의 분자적 진화 및 확산에 IS26 Mobile Element의 개입)

  • Kim, Jung-Min;Shin, Haeng-Seop;Cho, Dong-Taek
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.3
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    • pp.263-271
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    • 2000
  • A clinical isolate of Klebsiella pneumoniae K7746 produced the extended-spectrum ${\beta}$-lactamase (ESBL) SHV-12. A 6.6 kb BamHI fragment containing the $bla_{SHV-12}$ gene of K7746 strain was cloned into pCRScriptCAM vector resulting in the recombinant plasmid p7746-Cl. The restriction map of 3.6 kb inserted DNA and sequences immediately surrounding $bla_{SHV-12}$ of p7746-C1 were homologous to plasmid pMPA2a carrying $bla_{SHV-2a}$. In addition, both $bla_{SHV-12}$ and $bla_{SHV-2a}$ were expressed from a common hybrid promoter made of the -35 region derived from the left inverted repeat of IS26 and the -10 region from the $bla_{SHV}$ promoter itself. The results indicate that $bla_{SHV-12}$ and $bla_{SHV-2a}$ may have evolved from a common ancestor in the sequential order of $bla_{SHV-2a}$ first, followed by $bla_{SHV-12}$. Furthermore, by the PCR mapping method using primers corresponding to the IS26 and $bla_{SHV}$, the association between IS26 and $bla_{SHV}$ was studied in 12 clinical isolates carrying $bla_{SHV-2a}$, 27 clinical isolates carrying $bla_{SHV-12}$, and 5 reference strains carrying $bla_{SHV-1}$ to $bla_{SHV-5}$. All 39 strains carrying $bla_{SHV-2a}$ or $bla_{SHV-12}$ were positive by the PCR, providing confirmative evidence that IS26 has been involved in the evolution and dissemination of $bla_{SHV-2a}$ and $bla_{SHV-12}$. But 5 reference strains carrying $bla_{SHV-1}$ to $bla_{SHV-5}$ were negative by the PCR. Therefore, we concluded that the molecular evolutionary pathway of $bla_{SHV-2a}$ and $bla_{SHV-12}$ may be different from that of other $bla_{SHV-ESBL}$, e.g., $bla_{SHV-2}$, $bla_{SHV-3}$, $bla_{SHV-4}$, and $bla_{SHV-5}$.

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Identification and Analysis of Putative Polyhydroxyalkanoate Synthase (PhaC) in Pseudomonas fluorescens

  • Lim, Ju Hyoung;Rhie, Ho-Gun;Kim, Jeong Nam
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1133-1140
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    • 2018
  • Pseudomonas fluorescens KLR101 was found to be capable of producing polyhydroxyalkanoate (PHA) using various sugars and fatty acids with carbon numbers ranging from 2 to 6. The PHA granules consisted mainly of a poly(3-hydroxybutyrate) homopolymer and/or poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer. Genomic DNA of P. fluorescens was fractionated and cloned into a lambda library, in which a 5.8-kb fragment that hybridized to a heterologous phaC probe from Ralstonia eutropha was identified. In vivo expression in Klebsiella aerogenes KC2671 (pUMS), restriction mapping, Southern hybridization experiments, and sequencing data revealed that PHA biosynthesis by P. fluorescens relied upon a polypeptide encoded by a 1,683-bp non-operonal ORF, which was preceded by a possible -24/-12 promoter and highly similar to DNA sequences of a gene encoding PHA synthase in the genus Pseudomonas. In vivo expression of the putative PHA synthase gene ($phaC_{Pf}$) in a recombinant Escherichia coli strain was investigated by using glucose and decanoate as substrates. E. coli (${phaC_{Pf}}^+$, pUMS) grown in medium containing glucose accumulated PHA granules consisting mainly of 3-hydroxybutyrate, whereas only a trace amount of 3-hydroxydecanoate was detected from an E. coli fadR mutant (${phaC_{Pf}}^+$) grown in medium containing decanoate. In vitro enzymatic assessment experiments showed that 3-hydroxybutyryl-CoA was efficiently used as a substrate of purified $PhaC_{Pf}$, suggesting that the putative PHA synthase of P. fluorescens utilizes mainly short-chain-length PHA precursors as a substrate.

Stable Isotope and Fluid Inclusion Studies of the Manjang Copper Mine, South Korea (만장동광산(萬藏銅鑛山)에 대(對)한 유체포유물(流體包有物) 및 안정동위원소분석(安定同位元素分析) 연구(硏究))

  • Kim, Kyu Han;Shin, Jeung Sook
    • Economic and Environmental Geology
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    • v.20 no.3
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    • pp.169-177
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    • 1987
  • The Manjang copper magnetite-fluorite orebodies are imbedded within the limestone beds of the Hwajonri Formation. The ore deposits are characterized by magnetite-fluorite bearing skarn orebody in the west orebody and copper sulfide veins of the central and main orebodies. This study includes fluid inclusion geothermometry, salinity analysis, stable isotope analysis, and application of phase rule to mineral associations in skarn ore. Ore minerals are closely associated with the skarn silicates such as garnet, wollastonite and epidote. Magnetite and fluorite are remarkable in the west orebody whereas chalcopyrite is dominate in the central and main orebodies where pyrite and pyrrhotite also appear as sulfide gangues. Homogenization temperature and salinity of fluid inclusions are measured ranging between $240^{\circ}C$ and $350^{\circ}C$, 6.3~12.9 wt. percent in quartz and $220^{\circ}C$ and $350^{\circ}C$, 8.5~9.9wt. percent in fluorite, respectively. This indicates that the filling temperature and salinity are higher in quartz than in fluorite with the tendency of both to be linearly decreased suggesting an attribution of meteoric water to the mineralization. $T-fo_2$ diagram in the Ca-Fe-Si system at 1 kb and $Xco_2$=0.02 shows that the mineral assemblages with decreasing temperature are andradite-hedenbergite-calcite, hedenbergite-andradite-quartz, magnetite-andradite-quartz, and magnetite-quartz-calcite, indicating that magnetite crystallizes mostly late skarn stage at lower temperature. According to the carbon and oxygen isotopic values of the host limestone and calcite in ores, the sourec of carbon might be mixture of host limestone and deep seated carbons. Sulfur isotope data imply that ore fluids be relatively homogeneous in sulfur isotopic composition, mainly derived from igneous source.

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Cloning, Sequencing and Expression of the Gene Encoding a Thermostable β-Xylosidase from Paenibacillus sp. DG-22 (Paenibacillus sp. DG-22로부터 열에 안정한 β-xylosidase를 암호화하는 유전자의 클로닝, 염기서열결정 및 발현)

  • Lee, Tae-Hyeong;Lee, Yong-Eok
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1197-1203
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    • 2007
  • A genomic DNA library of the bacterium Paenibacillus sp. DG-22 was constructed and the ${\beta}-xylosi-dase-positive$ clones were identified using the fluorogenic substrate $4-methylumbelliferyl-{\beta}-D-xylopyr-anoside$ $({\beta}MUX)$. A recombinant plasmid was isolated from the clone and 4.3-kb inserted DNA was sequenced. The ${\beta}-xylosidase$ gene (xylA) was comprised of a 2,106 bp open reading frame (ORF) en-coding 701 amino acids with a molecular weight of 78,710 dalton and a pI of 5.0. The deduced amino acid sequence of the xylA gene product had significant similarity with ${\beta}-xylosidases$ classified into family 52 of glycosyl hydrolases. The xylA gene was subcloned into the pQE60 expression vector to fuse with six histidine-tag. The recombinant ${\beta}-xylosidase$ $(XylA-H_6)$ was purified to homogeneity by heat-treatment and immobilized metal affinity chromatography. The pH and temperature optima of the $XylA-H_6$ enzyme were pH 5.5-6.0 and $60^{\circ}C$, respectively.

Comparative Studies between Chungju and Seosan Groups (충주층군(忠州層群)과 서산층군(瑞山層群)의 비교연구(比較硏究))

  • Na, Ki Chang;Kim, Hyung Shik;Lee, Dong Jin;Lee, Sang Hun
    • Economic and Environmental Geology
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    • v.15 no.4
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    • pp.177-188
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    • 1982
  • The Chungju and Seosan Groups have been known usually as Precambrian formations in Korea. But their relative and absolute ages have been controvericial problem in relation with other geologic system such as so-called Ogcheon and Yeoncheon Systems in Korea. This study has mainly focused on the corelation of the Chungju Group with the Seosan Group in their stratigraphy, structure, metamorphism, and iron ore deposits. In the process of study, the auther surveyed and reclassified the Chungju and Seosan Groups and corelated with Gyeonggi and Ogch cheon metamorphic belts and got some new data. The Chungju iron-bearing formations showing transtitional relation with the Gyeonggi Gneiss Complex and the Jangamri Formation consisting mainly of pebble bearing calcarious phyllite, should be seperated from the Gyemyeongsan formation which is mainly composed of metavolcanic rocks. The Jangamri Formation and the coaly phyllite, which can be corelated respectively with the Hwaggangri Formation and Changri Formation in Ogcheon Group, are repeated in the Gyemyeonsan and Munjuri Formations with the overturned anticlinal folding(F1). So the Chungju Group which was defined as an indipendant geologic unit from the Ogcheon Group should be limited only on the Chungju iron Formation. The Seosan Group can be classified stratigraphically such as Seosan Formation consisting of iron-bearing quartzite and mica schist, Daesan Formation overlying unconformably on the Seosan Formation and Gyeonggi Gneiss Complex. Taean Formation overlying unconformably on the Daesan Formation should be seperated from Seosan Group. There are many similarity in the stratigrphy, structure, and metamorphic facies between Chungju and Seosan Groups exept the metavolcanic rocks in the Gyemyeongsan and Munjuri Formations and the pebble bearing calcareous phyllite in the Jangamri Formation. The two Groups were deformed with two kinds of differant stages, the first shows $N30^{\circ}-40^{\circ}E$ trend of fold axis, the second $N70^{\circ}-80^{\circ}W$ respectively. The Seosan Formation, which is the lowest formation in Seosan Group and bearing the iron formation, was metamorphosed at 2500 m. y. before. These age is similar with the metamorphic age of Gyeonggi metamorphic belt and with the age of Algoman and Kenoran Orogenies which devide the Precambrian into Archean and Proterozoic Era. So the Seosan Formation, which is included in some migmatitic rocks of Gyeonggi Gneiss Complex, is the oldest formation in Korea and can be corelated with the Anshan Group which bears the oldest iron formation in China. The metamorphic facies of the Precambrian metamorphism in Seosan area is simillar with that of Chungju area, showing high temperature-low pressure amphibolite facies which is corelated with the Gyeonggi metamorphic belt, the oldest metamorphic belt in Korea ($650^{\circ}-680^{\circ}C$, 3.2-4.4 Kb). The high temperature intermediate pressure amphibolite facies in Seosan area with the low temperature-intermediate presure greenschist facies of Taean formation is corelated with that of Ogcheon Group ($590^{\circ}-640^{\circ}$ C, 5.2-6.3 Kb). The Chungju and Seosan iron formations were deposited in Archean, showing geochemical composition of Precambrian iron formations. The Chungju iron formation was mainly formed by the chemical precipitation, on the other hand, the Seosan iron formation was formed by alternated action of chemical and detrital depositions.

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Cell Surface Display of Cycloinulooligosaccharide Fructanotransferase Gene in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 Cycloinulooligosaccharide Fructanotransferase 유전자의 표층 발현)

  • Kim, Hyun-Jin;Lee, Jae-Hyung;Kim, Hyun-Chul;Kim, Yeon-Hee;Kwon, Hyun-Ju;Nam, Soo-Wan
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.241-247
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    • 2007
  • The cycloinulooligosaccharide fructanotransferase (CFTase) gene (cft) from Paenibacillus macerans was subcloned into the surface display vector, pCTcon (GAL1 promoter). The constructed plasmid, pCTECFTN (9.0 kb) was introduced to S. cerevisiae EBY100 cell and then east transformants were selected on the synthetic defined medium lacking uracil and on the inulin containing medium. The surface display of CFTase was confirmed by immunofluorescence microscopy and its enzymatic ability to form cycloinulooligosaccharides(cyclofructans, CFs) from inulin. The total activity of the CFTase was reached about 5.52 unit/1 by cultivation of yeast transformant on YPDG medium. The optimized conditions determined were as follows; pH, 8.0; temperature, $50^{\circ}C$ ; substrate concentration, 5%; inulin source, Jerusalem artichoke. By the reaction with inulin, CFs consisting of cycloinulohexaose (CF6), cycloinuloheptaose (CF7), and cycloinulooctaose (CF8) were produced and CF6 was the major product.

Jiri Mountain, Korea : A Window into the Deep Crust (지리산 : 지각 깊은 곳을 들여다보는 창문)

  • Song, Yong-Sun;Park, Kye-Hun
    • The Journal of the Petrological Society of Korea
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    • v.26 no.4
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    • pp.385-398
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    • 2017
  • Jiri Mountain lies in the southwestern portion of the Yeongnam massif, which is one of the Precambrian basement massifs of the Korean Peninsular, consisting essentially of high-grade metamorphic rocks. The geology of the area mainly consists of Paleoproterozoic metasedimentary migmatitic gneisses, granitic gneisses which are classified into granitic gneiss, (K-feldspar porphyroblastic) granitic gneiss and quartzo-feldspathic gneiss, charnockite and anorthosite based on their occurrence and petrographic characteristics. The ages obtained from these rocks mainly span a narrow range between ca. 1,876 and 1,856 Ma although inherited cores of zircons from massive granite gneiss yielded much older age spectrum (>2,029 Ma). The age of major metamorphism is ca. 1850-1840 Ma and the metamorphic condition obtained from mineral assemblages and geothermobarometers is about 4-6 kb and up to $700-750^{\circ}C$. These results indicate that in the area intense granitic magmatism and metamorphism occurred in the deep crust during Paleoproterozoic orogeny. Some younger age of charnockite (1,856-1,865 Ma) and anorthosite (1,861-1,862 Ma) might indicate the beginning of intraplate rifting leading to felsic and mafic magmatism just after the orogeny. In conclusion, the rocks in the Jiri Mountain area which formed at a mid to deep crustal zone provide us windows into the deep crust.

Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii

  • Son, Eui-Sun;Song, Kyoung-Ju;Shin, Jong-Chul;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.133-141
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    • 2001
  • A cDNA of 1.1 kb comprising the gene encoding the peroxiredoxin of Toxo-plasma gondii(TgPrx) has been cloned. The open reading frame of 591 Up was translated into a protein of 196 amino acids with a molecular mass of 25 kDa. Conserved 2 cysteine domains of Phe-Val-Cys-Pro and Glu-Val-Cys-Pro indicated TgPrx belonged to 2-Cys Prx families. TgPrx showed the highest homology with that of Arabidopsis thaliana by 53.9% followed by Entamoeba histolytica with 39.5% by the amino acid sequence alignment. Polyclonal antibody against recombinant TgPrx detected 25 kDa band in T. gondii without binding to host cell proteins TgPrx was located in the cytoplasm of T. gondii extracellularly or intracellularly by immunofluorescence assay. The expression of TgPrx was increased as early as 30 min after the treatment with artemisinin in the intracellular stage, while no changes in those of host Prx I and TgSOD. This result implies that TgPrx may function as an antioxidant protecting the cell from the attack of reactive oxygen intermediates. It is also suggested that TgPrx is a possible target of chemotherapy.

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Complete Mitochondrial Genome Sequences of Korean Phytophthora infestans Isolates and Comparative Analysis of Mitochondrial Haplotypes

  • Seo, Jin-Hee;Choi, Jang-Gyu;Park, Hyun-Jin;Cho, Ji-Hong;Park, Young-Eun;Im, Ju-Sung;Hong, Su-Young;Cho, Kwang-Soo
    • The Plant Pathology Journal
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    • v.38 no.5
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    • pp.541-549
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    • 2022
  • Potato late blight caused by Phytophthora infestans is a destructive disease in Korea. To elucidate the genomic variation of the mitochondrial (mt) genome, we assembled its complete mt genome and compared its sequence among different haplotypes. The mt genome sequences of four Korean P. infestans isolates were revealed by Illumina HiSeq. The size of the circular mt genome of the four major genotypes, KR_1_A1, KR_2_A2, SIB-1, and US-11, was 39,872, 39,836, 39,872, and 39,840 bp, respectively. All genotypes contained the same 61 genes in the same order, comprising two RNA-encoding genes, 16 ribosomal genes, 25 transfer RNA, 17 genes encoding electron transport and ATP synthesis, 11 open reading frames of unknown function, and one protein import-related gene, tatC. The coding region comprised 91% of the genome, and GC content was 22.3%. The haplotypes were further analyzed based on sequence polymorphism at two hypervariable regions (HVRi), carrying a 2 kb insertion/deletion sequence, and HVRii, carrying 36 bp variable number tandem repeats (VNTRs). All four genotypes carried the 2 kb insertion/deletion sequence in HVRi, whereas HVRii had two VNTRs in KR_1_A1 and SIB-1 but three VNTRs in US-11 and KR_2_A2. Minimal spanning network and phylogenetic analysis based on 5,814 bp of mtDNA sequences from five loci, KR_1_A1 and SIB-1 were classified as IIa-6 haplotype, and isolates KR_1_A2 and US-11 as haplotypes IIa-5 and IIb-2, respectively. mtDNA sequences of KR_1_A1 and SIB-1 shared 100% sequence identity, and both were 99.9% similar to those of KR_2_A2 and US-11.

Characteristics of Glucose-6-phosphate Dehydrogenase from Leuconostoc mesenteroides (Leuconostoc mesenteroides에서 分離한 Glucose-6-phosphate Dehydrogenase의 特性)

  • Byun Si Myung;Yang Do Choi;Moon H. Han
    • Journal of the Korean Chemical Society
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    • v.23 no.4
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    • pp.248-258
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    • 1979
  • Glucose 6-phosphate dehydrogenase of Leuconostoc mesenteroides which was purifid by an affinity chromatography was studied on the characterization, kinetics and chemical modification. The apparent molecular weight of the enzyme was 112,000 by the gel filtration method of Sephadex G-200 column. The optimum temperature of $NAD^+$-linked reation was 50$^{circ}C$ and the activation energy and the heat of inactivation were 8.36 kcal/mole and -58.2kcal/mole, respectively. The steady state kinetic study showed KG6P, Kemp, and CX KNADP to be 76.9 PM, 7.46${\mu}M$ and 7.14 ${\mu}M$, respectively, and KGGP, KNAD,and aKNm to be 53.7${\mu}M$, 115.2${\mu}M$ and 702.2${\mu}M$ for the $NAD^+$-linked reaction at pH 7.8, optimum pH. The pH dependent kinetic constants suggested that the two ionizing groups whose pKa is 7.2 .and pKb is 9.0-9.6 were involved in the enzyme-substrate interaction. Evidence by photooxidation and carboxymethylation of the enzyme suggested that the imidazole group of histidine with pKa group may participate in the catalytic site.

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