• Title/Summary/Keyword: Insertion strain

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Identification of a Novel Genetic Locus Affecting ptsG Expression in Escherichia coli

  • Shin Dong-Woo;Lee Sang-Mi;Shin Yu-Rae;Ryu Sang-Ryeol
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.795-798
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    • 2006
  • The phosphoenolpyruvate-dependent carbohydrate phosphotransferase system (PTS) is responsible for the simultaneous transfer and phosphorylation of various carbon sources in Escherichia coli. The ptsG gene encoding the enzyme $IICB^{Glc}$, the membrane component of the glucose-specific PTS, is repressed by Mlc and activated by the CRP cAMP complex; various other factors, such as Fis, FruR, and ArcA, are also known to be involved in ptsG regulation. Thus, in an attempt to discover a novel gene affecting the regulation of ptsG, a mutant with a decreased ptsG transcription in the presence of glucose compared with the wild-type strain was screened using transposon random mutagenesis. The mutant was found to have a transposon insertion in yhjV, a putative gene encoding a transporter protein whose function is yet unknown.

Analysis of polymorphic region of GAM-1 gene in Plasmodium vivax Korean isolates

  • Kho, Weon-Gyu;Chung, Joon-Yong;Hwang, Ui-Wook;Chun, Jin-Ho;Park, Yeong-Hong;Chung, Woo-Chul
    • Parasites, Hosts and Diseases
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    • v.39 no.4
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    • pp.313-318
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    • 2001
  • The identification , characterization and quantification of Plasmodium sp. genetic polymorphism are becoming increasingly important in the vaccine development. We investigated polymorphism of Plasmodium vivax GAM-1 (PvGAM-1) gene in 30 Korean isolates. The polymorphic region of the PvGAM-1 gene, corresponding to nt 3792-4029, was amplified using polymerase chain reaction (PCR) followed by sequencing. All of the P. viuax Korean isolates were one type of GAM-1 gene, which were identical to that of the Belem strain. It is suggested that PvGAM-1 could not be used as a genetic marker for identifying or classifying P. vivax Korean isolates. It revealed that the polymorphic pattern as acquired basically by duplication and modification or deletion event of a 33 bp-motif fragment ended by poly guanine (G) and that there were at least three complete and one partial 33 Up-motif sequences within the polymorphic region in the longest cases such as those of South Korean and Belem isolates. In addition, we clustered P. vivax isolates with parsimonious criteria on the basis of PvGAM- 1 polymorphic patterns (insertion/deletion patterns) .

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Temperature and the Interfacial Buffer Layer Effects on the Nanostructure in the Copper (II) Phthalocyanine: Fullerene Bulk Heterojunction

  • Kim, Hyo Jung;Kim, Jang-Joo;Jeon, Taeyeol;Kong, Ki Won;Lee, Hyun Hwi
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.275.1-275.1
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    • 2014
  • The effects of the interfacial buffer layer and temperature on the organic bulk heterojunction (BHJ) nanostructures of copper phthalocyanine (CuPc) and fullerene (C60) systems were investigated using real time in-situ x-ray scattering. In the CuPc:C60 BHJ structures, standing-on configured ${\gamma}$-CuPc phase was formed by co-deposition of CuPc and C60. Once formed ${\gamma}$-phase was thermally stable during the annealing upon $180^{\circ}C$. Meanwhile, the insertion of CuI buffer layer prior to deposition of the CuPc:C60 BHJ layer induced lying-down configured CuPc crystals in the BHJ layer. The lying CuPc peak intensity and the lattice parameter were increased by the thermal annealing. This increment of the intensity seemed to be related to the strain at the interface between CuPc:C60 and CuI, which was proportional to the enhancement of the power conversion efficiency of the device.

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Electrochemistry Characteristics of $Li_4Ti_5O_{12}$ Anode Electrode for Li-ion Battery (리튬전지용 $Li_4Ti_5O_{12}$ 음극전극의 전기화학적 특성)

  • Oh, Mi-Hyun;Kim, Han-Joo;Kim, Young-Jae;Son, Won-Keun;Lim, Kee-Joe;Park, Soo-Gil
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2005.11a
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    • pp.340-341
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    • 2005
  • Lithium titanium oxide as anode material for energy storage prepared by novel synthesis method. $Li_4Ti_5O_{12}$ based spinel-framework structures are of great interest material for lithium-ion batteries. We describe here $Li_4Ti_5O_{12}$ a zero-strain insertion material was prepared by novel sol-gel method and by high energy ball milling (HEBM) of precursor to from nanocrystalline phases. According to the X-ray diffraction and scanning electron microscopy analysis, uniformly distributed $Li_4Ti_5O_{12}$ particles with grain sizes of 100nm were synthesized. Lithium cells, consisting of $Li_4Ti_5O_{12}$ anode and lithium cathode showed the 173 mAh/g in the range of 1.0 $\sim$ 3.0 V. Furthermore, the crystalline structure of $Li_4Ti_5O_{12}$ didn't transfer during the lithium intercalation and deintercalation process.

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Molecular Colning and Ewpression of the $\alpha$-L-Arabinofuranosidase Gene of Bacillus stearothermophilus in Escherichia coli (Bacillus stearothermophilus로부터 $\alpha$-L-Arabinofuranosidase 유전자의 클로닝 및 Escherichia coli에서의 발현)

  • Eom, Soo-Jung;Kim, Hee-Sun;Cho, Ssang-Goo;Choi, Yong-Jin
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.607-613
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    • 1994
  • The Bacillus stearothermophilus arfI gene encoding a-arabinofuranosidase was isolated from the genomic library, cloned into pBR322, and subsequently transferred into the Escherichia coli HB101. The recombinant E. coli was selected from approximately 10,000 transformants screened by making use of its ability to produce a yellow pigment around the colony on the selective medium supplemented with p-nitrophenyl-$\alpha$-L-arabinofuranoside (pNPAf), a chromogenic substrate. The functional clone was found to harbor a recombinant plasmid, pKMG11 with an insertion of about 5 kb derived from the B. stearothermophilus chromosomal DNA. Identity of the arfI gene on the insert DNA was confirmed by a zymogram with 4-methylumbelliferyl-$\alpha$-L-arabinofuranoside as the enzyme substrate. The $\alpha$-arabinofuranosidase from the recombinant E. coli strain showed very high substrate specificity; the enzyme displayed high activity only with pNPAf among many other p- or $o$-nitrophenyl derivatives of several sugars, and acted only on arabinoxylan among various natural arabinose containing polysaccharides tested.

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Development and evaluation of semi-nested PCR for detection of the variable lipoprotein haemagglutinin (vlhA) gene of Mycoplasma Synoviae in chicken

  • Pohuang, Tawatchai;Phuektes, Patchara;Junnu, Sucheeva
    • Korean Journal of Veterinary Research
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    • v.60 no.3
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    • pp.109-116
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    • 2020
  • This study aimed to develop a semi-nested polymerase chain reaction assay for the direct detection of Mycoplasma synoviae (M. synoviae) from clinical samples using three newly designed oligonucleotide primers specific to the variable lipoprotein haemagglutinin (vlhA) gene and differentiate M. synoviae field strains based on a nucleotide deletion or the insertion of the proline-rich repeat (PRR) region of the vlhA gene. The developed semi-nested polymerase chain reaction (PCR) assay revealed positive results in 12 out of 100 clinical samples collected from chickens showing lameness and joint swelling. Six positive samples were selected randomly for sequencing, and sequence analysis revealed 96.3-100% nucleotide identities compared to the reference sequences. Phylogenetic analysis showed that sequences of the strains in this study were closely related to WVU1853 (Spain), CK.MS.UDL.PK.2014.2 (Pakistan), and F10-2AS (USA) strains, but they were distinct from the M. synoviae-H vaccine strain sequence. M. synoviae obtained from these samples were identified as types A and C with a length of 38 and 32 amino acids, respectively. These results indicated that the specific and sensitive semi-nested PCR could be a useful diagnostic tool for the direct identification of clinical samples, and the sequence analysis of the partial vlhA gene can be useful for typing M. Synoviae.

Magnetic Properties of Multiferroic $BiFeO_3/BaTiO_3$ Bi-layer Thin Films

  • Yang, P.;Byun, S.H.;Kim, K.M.;Lee, Y.H.;Lee, J.Y.;Zhu, J.S.;Lee, H.Y.
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.06a
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    • pp.318-319
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    • 2008
  • In this article, magnetic properties of multiferroic bi-layer $BiFeO_3$ (BFO)/$BaTiO_3$ (BTO) thin films were studied. It was found that the magnetization increased by the insertion of BTO buffer layer even though the interfacial stress was slightly relaxed, which indicated a coupling between the ferroelectric and ferromagnetic orders. Furthermore, with slightly increase of BFO film thickness, both BFO and BFO/BTO bi-layer films showed anisotropic magnetic properties with higher in-plane magnetization than the values measured out-of-plane. These are attributable to strain constraint effect at the interface.

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Regulation of SoxR, the superoxide-sensory regulator in Escherichia coli.

  • Lee Joon-Hee;Koo Mi-Sun;Yeo Won-Sik;Roe Jung-Hye
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.24-31
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    • 2000
  • In order to find out SoxR-reducing system in E. coli, we generated Tn10-insertion mutants and screened for constitutive expression of SoxS in a soxS-lacZ fusion strain. One mutation was mapped in rseB, a gene in rseABC (Regulation of SigmaE) operon. The constitutive soxS-expressing phenotype was due to the polar effect on the downstream gene, rseC. RseC is likely to function as a component of SoxR reduction system because SoxR was kept in oxidized form to activate soxS expression in rseC mutant. RseC is an integral membrane protein with an N-terminal cysteine-rich domain in the cytoplasm. The functionally critical cysteines were determined by substitution mutagenesis. The truncated N-terminal domain of RseC reduced the soxS transcription by $50\%$ as judged by in vitro transcription assay. Currently RseC is believed to be a reducing factor for SoxR. However, the mechanism for the reduction needs further investigation.

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Cloning and Expression of a Full-Length Glutamate Decarboxylase Gene from Lactobacillus plantarum

  • Park, Ki-Bum;Oh, Suk-Heung
    • Preventive Nutrition and Food Science
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    • v.9 no.4
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    • pp.324-329
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    • 2004
  • In order to investigate the molecular mechanism of $\gamma$-aminobutyric acid (GABA) production in lactic acid bacteria, we cloned a glutamate decarboxylase (GAD) gene from Lactobacillus plantarum using polymerase chain reaction (PCR). One PCR product DNA was obtained and inserted into a TA cloning vector with a T7 promoter. The recombinant plasmid was used to transform E. coli. The insertion of the product was con­firmed by EcoRI digestion of the plasmid purified from the transformed E. coli. Nucleotide sequence analysis showed that the insert is a full-length Lactobacillus plantarum GAD and that the sequence is $100\%$ and $72\%$ identical to the regions of Lactobacillus plantarum GAD and Lactococcus lactis GAD sequences deposited in GenBank, accession nos: NP786643 and NP267446, respectively. The amino acid sequence deduced from the cloned Lactobacillus plantarum GAD gene showed $100\%$ and $68\%$ identities to the GAD sequences deduced from the genes of the NP786643 and NP267446, respectively. To express the GAD protein in E. coli, an expression vector with the GAD gene (pkk/GAD) was constructed and used to transform the UT481 E. coli strain and the expression was confirmed by analyzing the enzyme activity. The Lactobacillus plantarum GAD gene obtained may facilitate the study of the molecular mechanisms regulating GABA metabolism in lactic acid bacteria.

Novel Synthesis Method and Electrochemical Characteristics of Lithium Titanium Oxide as Anode Material for Lithium Secondary Battery

  • Kim Han-Joo;Park Soo-Gil
    • KIEE International Transactions on Electrophysics and Applications
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    • v.5C no.3
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    • pp.119-123
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    • 2005
  • Lithium titanium oxide as anode material for energy storage prepared by novel synthesis method. Li$_{4}$Ti$_{5}$O$_{12}$ based spinel-framework structures are of great interest material for lithium-ion batteries. We describe here Li$_{4}$Ti$_{5}$O$_{12}$ a zero-strain insertion material was prepared by novel sol-gel method and by high energy ball milling (HEBM) of precursor to from nanocrystalline phases. According to the X-ray diffraction and scanning electron microscopy analysis, uniformly distributed Li$_{4}$ Ti$_{5}$O$_{12}$ particles with grain sizes of 100nm were synthesized. Lithium cells, consisting of Li$_{4}$ Ti$_{5}$O$_{12}$ anode and lithium cathode showed the 173 mAh/g in the range of 1.0 $\~$ 3.0 V. Furthermore, the crystalline structure of Li$_{4}$ Ti$_{5}$O$_{12}$ didn't transform during the lithium intercalation and deintercalation process.