• Title/Summary/Keyword: lac I

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Activity of virE promoter in Agrobacterium tumefaciens A348 (Agrobacterium tumefaciens A348에서 virE 프로모터의 활성)

  • 음진성
    • Journal of Plant Biology
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    • v.34 no.4
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    • pp.331-339
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    • 1991
  • To elucidate the regulatory mechanism of virE operon from vir regions (virA, virB, virC, virD, virG, virE) of pTiA6 which have been known to be essential for efficient crown gall tumorigenesis in plants, the activity of the truncated virE, promoter was analyzed. pSM358cd, a recombinant plasmid in which virE :: Tn3-HoHo1 (Tn3-promoterless lacZ) was cloned into SalI site of pVK102, was digested with SalI, and virE :: Tn3-HoHo1 was seperated from pVK102. To construct the truncted virE recombinant plasmids (pJS031, pJS051, pJS102, pJS201, pJS301), 5'-end of vireE promoter was deleted with BAL31 and cloned into pVK102 and then transferred into a. tumefaciens A348(pTiA6). According to the activity of the truncated virE promoter in recombinant plasmids, they were classified into two groups, pJS031, pJS051, pJS101 and pJS201 belong to a functional group and pJS301 is a non-functional. The size of deleted nucleotides of pJS201 and pJS301 seemed to be about 130 nucleotides and about 250 nucleotides from 5'-end of virE promoter, respectively. Hence it was thought that the essential site of the virE promoter was located between about 130th nucleotide and 250th nucleotide from 5'-end of the virE promoter.

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Eclipse Spectrum of Her X-1 Observed by ASCA in the Low Intensity State

  • Choi, C.S.;Seon, K.I.
    • Bulletin of the Korean Space Science Society
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    • 1995.10a
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    • pp.30-30
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    • 1995
  • We present the results of analysis on the X-ray observations of the binary X-ray pulsar Her X-I. made with ASCA/SIS on August 13-14. 1993. An eclipse transition from ingress to egress was fully covered by the observations. The main findings are as follows; (1) a model of power-law plus black-body is required to interpret the entire eclipse spectrum. and the black-body component appears at < 0.7 keV. (2) the power-law continuum which has photon index ${\alpha}\;=\;{0.84^{\;+0.14}}_{\;-0.19}$ is very similar to that of detected by Ginga/LAC (${\alpha}\;=\;0.80\;{\pm}\;0.04$), (3) the calculated eclipse flux of $2^{-10}\;keV.{\;}~{\;}1.8{\pm}10^{-11}{\;}ergs{\;}cm^{-2}s^{-1}$, is consistent with the Ginga observation carried out in the high intensity state ~2.0{\pm}10^{-11}{\;}ergs{\;}cm^{-2}s^{-1}$, (4) there is no significant absorption feature. and an upper limit of the aborption column $NH{\;}\leq{\;}3{\pm}10^{20}\;cm^{-2}$ is determined at the 90% confidence limit. Based on these results, we suggest that extended matter surrounding the binary system should be existed persistently with stable conditions, and scattering of the source continuum by the matter is responsible for the eclipse emission.ission.

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Characterization of the scr Gene Cluster Involved! in Sucrose Utilization in Bifidobacterium longum (Bifidobacterium longum의 Sucrose 대사 관련 scr 유전자군의 특성 규명)

  • 권태연;이종훈
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.199-205
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    • 2004
  • The nucleotide sequence of 8.6-kb EcoRI fragment containing sucrose phosphorylase gene isolated from Bifidobacterium longum SJ32 was determined. It was found that the fragment contained five open reading frames including the gene cluster for sucrose utilization such as a sucrose phosphorylase (ScrP), a sucrose transporter (ScrT), and a GalR-LacI-type transcriptional regulator (ScrR) identified by amino acid homology. Each gene showed over 94% amino acid homology among various B. longum strains. Genomic organization of the gene cluster is the same as those of other strains of B. longum but different from that of B. lactis. In spite of high homology of each gene among B. longum strains, the difference of flanking sequences of the gene cluster between strains SJ32 and NCC2705 insinuates the horizontal transfer of scrPTR between B. longum strains. The increase of sucrose phosphorylase activity in heterologous E. coli system by the co-expression of scrT with scrP against the single expression of scrP was measured. It seems to be the result of sucrose uptake increment by scrT in the host and is an indirect evidence that scrT is the gene for sucrose transport. The existence of multiple sucrose uptake systems in B. longum is supposed from the findings of several genes besides scrPTR involved in sucrose uptake in the genome of B. longum NCC2705.

Construction of New Transfer Vector of Nuclear Polyhedrosis Virus of the Silkworm, Bombyx mori (누에 핵다각체병 바이러스를 이용한 새로운 전이 벡터의 제작)

  • 우수동;김우진;진병래;강석권
    • Journal of Sericultural and Entomological Science
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    • v.37 no.1
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    • pp.46-51
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    • 1995
  • In order to develope baculovirus expression vector system, we constructed new transfer vector of nuclear polyhedrosis virus of the silkworm, Bombyx mori. The promoter region containing only adenine of translation start codon of polyhedrin gene was cloned by polymerase chain reaction technique. And the 5' and 3' leader regions of polyhedrin gene was sequentially cloned. The polyhedrin coding gene was deleted from the +2 to the +597 position. As the result, we constructed new transfer vector which has EcoRI, SacI and KpnI sites for the cloning sites of foreign gene. New transfer vector was named as pBmKSKl. Escherichia coli $\beta$-galactosidase gene as foreign gene was inserted into pBmKSKl, under the control of the polyhedrin promoter and expressed in B. mori cells. The result showed that the new transfer vector pBmKSK1 is functional.

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Regulation of Photosynthesis Genes (puf, puc, puhA, bchC, bchE, bchF, and bchI) in Rhodobacter sphaeroides (Rhodobacter sphaeroides에서의 광합성유전자(puf, puc, puhA, bchC, bchE, bchF와 bchI)의 발현조절)

  • Ko, In-Jeong;Kim, Yong-Jin;Lee, Jin-Mok;Shin, Sun-Joo;Oh, Jeong-Il
    • Journal of Life Science
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    • v.16 no.4
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    • pp.632-639
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    • 2006
  • Here we examined the expression patterns and regulation of seven photosynthesis (PS) genes (puf, puc, puhA, bchC, bchE, bchF, and bchI) in the anoxygenic photosynthetic bacterium, Rhodobacter sphaeroides, based on lacZ reporter gene assay. Expression of the tested PS genes, except puhA and bchI, were strongly induced in R. sphaeroides grown under anaerobic conditions relative to that under aerobic conditions. The puhA and bchI genes appear to form the operons together with bchFNBHLM-RSP0290 and crtA, respectively. Expression of the puf, puc, and bchCXYZ operons in R. sphaeroides grown photosynthetically was proportional to the incident light intensity, whereas that of bchFNBHLM(RSP0290-puhA) was inversely related to light intensity. Expression of bchEJG was lowest under medium-light photosynthetic conditions $(10\;W/m^2)$ and highest under high light conditions $(100\;W/m^2)$. The regulation of PS genes by the three major regulatory systems involved in oxygen- and light-sensing in R. sphaeroides is as following: puf and bchC are regulated by both the PpsR repressor and the PrrBA two-component system. The puc operon is under control of PpsR, FnrL, and PrrBA system. Expression of bchE is controlled by FnrL and PrrBA two-component system, whereas bchF is regulated exclusively by PpsR. It was demonstrated that the PpsR repressor is responsible for high-light repression of bchF and that FnrL might be involved in perceiving the cellular redox state in addition to sensing $O_2$ itself.

Seismic Facies Classification of Igneous Bodies in the Gunsan Basin, Yellow Sea, Korea (탄성파 반사상에 따른 서해 군산분지 화성암 분류)

  • Yun-Hui Je;Ha-Young Sim;Hoon-Young Song;Sung-Ho Choi;Gi-Bom Kim
    • Journal of the Korean earth science society
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    • v.45 no.2
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    • pp.136-146
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    • 2024
  • This paper introduces the seismic facies classification and mapping of igneous bodies found in the sedimentary sequences of the Yellow Sea shelf area of Korea. In the research area, six extrusive and three intrusive types of igneous bodies were found in the Late Cretaceous, Eocene, Early Miocene, and Quaternary sedimentary sequences of the northeastern, southwestern and southeastern sags of the Gunsan Basin. Extrusive igneous bodies include the following six facies: (1) monogenetic volcano (E.mono) showing cone-shape external geometry with height less than 200 m, which may have originated from a single monogenetic eruption; (2) complex volcano (E.comp) marked by clustered monogenetic cones with height less than 500 m; (3) stratovolcano (E.strato) referring to internally stratified lofty volcanic edifices with height greater than 1 km and diameter more than 15 km; (4) fissure volcanics (E.fissure) marked by high-amplitude and discontinuous reflectors in association with normal faults that cut the acoustic basement; (5) maar-diatreme (E.maar) referring to gentle-sloped low-profile volcanic edifices with less than 2 km-wide vent-shape zones inside; and (6) hydrothermal vents (E.vent) marked by upright pipe-shape or funnel-shape structures disturbing sedimentary sequence with diameter less than 2 km. Intrusive igneous bodies include the following three facies: (1) dike and sill (I.dike/sill) showing variable horizontal, step-wise, or saucer-shaped intrusive geometries; (2) stock (I.stock) marked by pillar- or horn-shaped bodies with a kilometer-wide intrusion diameter; and (3) batholith and laccoliths (I.batho/lac) which refer to gigantic intrusive bodies that broadly deformed the overlying sedimentary sequence.

Cloning and Characterization of Autonomously Replicating Sequence(ARS) from Kluyveromyces fragilis

  • HONG, SOON-DUCK;JONG-GUK KIM;TAKUYA NAGAMATSU;JOO-HYUN NAM;DONG-SUN LEE;SANG-YONG LEE;SUN-HWA HA
    • Journal of Microbiology and Biotechnology
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    • v.3 no.1
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    • pp.6-11
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    • 1993
  • An autonomously replicating sequence (Kf-ARS1) of Kluyveromyces fragilis was cloned from the genomic library which was constructed using pHN134 as a cloning vector to make a new host-vector system for the production of heterologous protein from K. fragilis as a host. The cloning vector pHN134 was composed of $Km^r, Ap^r$ and multiple cloning site in LacZ . A clone carrying Kf-ARS1 was isolated and the recombinant plasmid was designated as pIKD102. The cloned fragment was 2.3 kb (EcoRI/EcoRI) in length. Subcloning experiment showed that the region for ARS activity was 1.5 kb (SalI/EcoRI) fragment. It was shown that the Kf-ARS1 was active in Saccharomyces cerevisiae and Kluyveromyces fragilis.

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Cytotoxicity of Cytosine Deaminase (CD) Adenoviral Vectors(AV) with a Promoter (L-plastin) for Epithelial Cancer Cells.

  • Chung, Injae;Jung, Kihwa;Deisseroth, Albert B.
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.80-80
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    • 1997
  • The object of this study was to develop a gene therapy strategy for ovarian cancer. We have previously shown that AV with a L-plastin (LP) promoter infects breast and ovarian cancer cells and expressed ${\beta}$-galactosidase cDNA in preference to normal fibroblast cells and hematopoietic cells. We now report on the cytotoxicity of Ad.LP.CD, an AV carrying a CD cDNA which converts the pro-drug, 5-Fluorocytosine (5-FC) into the toxic drug 5-Fluorouracil (5-FU). Infection of Ad.LP.CD into either 293 cells or ovarian cancer cells generated the functional CD as measured by HPLC analysis. Using a ratio of AV to OVCAR3 cell of 100 and a 5-FC concentration of 100 ${\mu}$M, we achieve an over 95 % of cell growth inhibition. We are using flow cytometry analysis for ${\beta}$ -galactosidase and ovarian cancer associated folate receptor to screen primary ascites samples for infectivity after infection with an adenoviral vector, i.e., Ad.LP.LacZ. This vector system may be of value in the treatment of microscopic disease of ovarian cancer in the peritoneal cavity.

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Cloning, Expression, and Characterization of DNA Polymerase from Hyperthermophilic Bacterium Aquifex pyrophilus

  • Choi, Jeong-Jin;Kwon, Suk-Tae
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1022-1030
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    • 2004
  • The gene encoding Aquifex pyrophilus (Apy) DNA polymerase was cloned and sequenced. The Apy DNA polymerase gene consists of 1,725 bp coding for a protein with 574 amino acid residues. The deduced amino acid sequence of Apy DNA. polymerase showed a high sequence homology to Escherichia coli DNA polymerase I-like DNA polymerases. It was deduced by amino acid sequence alignment that Apy DNA polymerase, like the Klenow fragment, has only the two domains, the $3'{\rightarrow}5'$ exonuclease domain and the $5'{\rightarrow}3'$ polymerase domain, containing the characteristic motifs. The Apy DNA polymerase gene was expressed under the control of T7lac promoter on the expression vector pET-22b(+) in E. coli. The expressed enzyme was purified by heat treatment, and Cibacron blue 3GA and $UNO^{TM}$ Q column chromatographies. The optimum pH of the purified enzyme was 7.5, and the optimal concentrations of KCl and $Mg^{2+}$ were 20 mM and 3 mM, respectively. Apy DNA polymerase contained a double strand-dependent $3'{\rightarrow}5'$ proofreading exonuclease activity, but lacked any detectable $5'{\rightarrow}3'$ exonuclease activity, which is consistent with its amino acid sequence. The somewhat lower thermostability of Apy DNA polymerase than the growth temperature of A. pyrophilus was analyzed by the comparison of amino acid composition and pressure effect.

Mapping of Gene Encoding Phospho-$\beta$-galactosidase from Lactobacillus casei and its Expression in Escherichea coli (Lactobacillus casei 의 Phospho-$\beta$-galactosidase 유전자의 지도작성과 Escherichia coli 내에서의 발현)

  • 박정희;문경희;민경희
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.539-545
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    • 1992
  • Recombinant plasmid pPLac15 determined both phosphoenolpyruvate-dependent phosphotransferase uptake of lactose and phospho-$\beta$-galactosidase (Moon et al., 1989). A restriction mapping of the pPLac15 was compiled with several restriction enzymes and a seriese of sub clones into pUC18 was constructed. From an analysis of the proteins produced by Escherichia coli cells of transformants containing each of the recombinant subclone plasmids, it was found that the gene for phospho-$\beta$-galactosidase in pUCI8 was expressed about 1.8-folds in E. coli.

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