• Title/Summary/Keyword: Rep. of Korea

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Molecular Interactions of a Replication Initiator Protein, RepA, with the Replication Origin of the Enterococcal Plasmid p703/5

  • Cha, Kyung-Il;Lim, Ki-Hong;Jang, Se-Hwan;Lim, Wang-Jin;Kim, Tae-Hyung;Chang, Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1841-1847
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    • 2007
  • We previously identified the origin of replication of p703/5, a small cryptic plasmid from the KBL703 strain of Enterococcus faecalis. The origin of replication contains putative regulatory cis-elements required for replication and a replication initiator (RepA) gene. The replicon of p703/5 is similar in its structural organization to theta-type plasmids, and RepA is homologous to a family of Rep proteins identified in several plasmids from Gram-positive bacteria. Here, we report molecular interactions between RepA and the replication origin of p703/5. DNase I footprinting using recombinant RepA together with electrophoretic mobility shift assays confirmed the binding of RepA to the replication origin of p703/5 via iterons and an inverted repeat. We also demonstrated the formation of RepA dimers and the different binding of RepA to the iteron and the inverted repeat using gel filtration chromatographic analysis, a chemical crosslinking assay, and electrophoretic mobility shift assays in the presence of guanidine hydrochloride. Our results suggest that RepA plays a regulatory role in the replication of the enterococcal plasmid p703/5 via mechanisms similar to those of typical iteroncarrying theta-type plasmids.

Frequency-stabilized Femtosecond Mode-locked Laser for Optical Frequency Metrology

  • Yoon, Tai-Hyun;Kim, Eok-Bong;Park, Seong-Tae
    • Journal of the Optical Society of Korea
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    • v.7 no.3
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    • pp.131-134
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    • 2003
  • We demonstrated an optical frequency synthesizer based on a femtosecond (fs) mode-locked Ti:sapphire (Ti:s) laser by simultaneously stabilizing the carrier-offset frequency, $f_{ceo}$, and repetition rate, $f_{ rep}$, referenced to the Cs atomic frequency standard. By using two wide-band digital phase-detectors we realized a phase-coherent link between $f_{rep} and f_{ceo} with the relation f_{ceo} = f_{AOM} 5/6f_{rep} ≡ 0, where f_{AOM} = 5/6f_{rep}$ is the phase-locked driving frequency of an acousto-optic modulator (AOM) in a self-referencing interferometer and $f_{rep}$ = 100 MHz. As a result, we could stabilize all components of the fs laser comb at once with an equal frequency separation $f_{rep}$ = 100 MHz with $f_{ceo}$ = 0. In our optical frequency synthesizer, the frequency of the nth component ($f_{n}$) is given exactly by the simple relation $f_n = nf_{rep}$, enabling us to use the fs laser comb as a frequency ruler in the optical frequency metrology.

Identification of Functionally Different Rat IgE in RBL-2H3 Exocytosis

  • Kim, Jin-Sub;Cho, Sungae;Joo, Kyoung-Hwan;Lee, Joon-Sang;Conrad, Daniel H.;Cho, Sung-Weon
    • IMMUNE NETWORK
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    • v.2 no.4
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    • pp.195-201
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    • 2002
  • Background: IgE is closely related to the development of allergies. However, the poor relationship between the specific IgE level and the severity of allergic diseases suggests the possibility of functionally different IgE isoforms. With this in mind, rat basophilic leukemia (RBL)-2H3 activation was analyzed with each type of rat IgE for two parameters, exocytosis and IL-4 mRNA production. RBL-2H3 has been well documented in the rat mucosal mast cell line. Methods: RBL-2H3 cells sensitized with each kind of rat IgE was activated by cross-linking FcRI with B5 (monoclonal anti-rat IgE mouse IgG antibodies). The RBL-2H3 exocytosis was measured by analyzing the ${\beta}$-hexosaminidase level, and the level of IL-4 mRNA synthesis was analyzed using semiquantitative RT-PCR. Rat IgE, which was produced by a parasite infection (REP), was prepared using either Paragonimus westermani metacercariae (REP-PW) or Anisakis simplex third stage larvae (REP-AS). A rat IgE prototype of IR162 was prepared by a peritoneal injection of immunocytoma. Results: The level of exocytosis showed a linear relationship with the rat IgE concentration when REP-PW or REP-AS was applied. However, it exhibited a biphasic response with IR162. In addition, the time course of heating at $56^{\circ}C$ illustrated the similarity between REP-PW and REP-AS, which differed from that of IR162. In contrast, the level of IL-4 mRNA synthesis in the RBL-2H3 cells with IR162 was comparable to that of either REP-PW or REP-AS. Conclusion: These results suggest that functionally different rat IgE isoforms exists in RBL-2H3 exocytosis.

Use of Reference Ear Plug to improve accuracy of lateral cephalograms generated from cone-beam computed tomography scans

  • Hwang, Hyeon-Shik;Lee, Kyung-Min;Uhm, Gi-Soo;Cho, Jin-Hyoung;McNamara, James A. Jr.
    • The korean journal of orthodontics
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    • v.43 no.2
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    • pp.54-61
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    • 2013
  • Objective: The purpose of this study was to evaluate the effectiveness of the use of Reference Ear Plug (REP) during cone-beam computed tomography (CBCT) scan for the generation of lateral cephalograms from CBCT scan data. Methods: Two CBCT scans were obtained from 33 adults. One CBCT scan was acquired using conventional methods, and the other scan was acquired with the use of REP. Virtual lateral cephalograms created from each CBCT image were traced and compared with tracings of the real cephalograms obtained from the same subject. Results: CBCT scan with REP resulted in a smaller discrepancy between real and virtual cephalograms. In comparing the real and virtual cephalograms, no measurements significantly differed from real cephalogram values in case of CBCT scan with REP, whereas many measurements significantly differed in the case of CBCT scan without REP. Conclusion: Measurements from CBCT-generated cephalograms are more similar to those from real cephalograms when REP are used during CBCT scan. Thus, the use of REP is suggested during CBCT scan to generate accurate virtual cephalograms from CBCT scan data.

Nucleotide Sequences of Rep and CAT Proteins encoded by Chloramphenicol-Resistance Plasmid pKH7 (클로람페니콜 내성 플라스미드 pKH7의 Rep 단백질과 CAT 단백질의 염기서열 분석)

  • 윤성준;이대운;김우구;신철교;임성환;문경호
    • YAKHAK HOEJI
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    • v.39 no.6
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    • pp.676-680
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    • 1995
  • The nucleotide sequence of Xbal-Mbol fragment of pKH7, a chloramphenicol-resistant($Cm^{r}$) plasmid isolated from multidrug-resistant S. aureus SA2, has been determined. Xbal-Mbol fragment of pKH7 was found to contain two ORFs. One ORF encoded Rap and the other encoded CAT protein. The deduced amino acid sequences of Rep and CAT of pKH7 were compared to those of pUB112 and pC221. Comparisons revealed that there was one amino acid difference in CAT between pKH7 and pUB112. CAT of pKH7 exhibited 98.6% amino acid identity to that of pC221. In case of Rep proteins, a slightly lower homology of 96.4% and 86.7% in amino acid sequences was observed between pKH7 and pUB112 and between pKH7 and pC221, respectively.

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The Development of Converting Program from Sealed Geological Model to Gmsh, COMSOL for Building Simulation Grid (시뮬레이션 격자구조 제작을 위한 Mesh 기반 지질솔리드모델의 Gmsh, COMSOL 변환 프로그램 개발)

  • Lee, Chang Won;Cho, Seong-Jun
    • Journal of the Korean earth science society
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    • v.38 no.1
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    • pp.80-90
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    • 2017
  • To build tetrahedra mesh for FEM numerical analysis, Boundary Representation (B-Rep) model is required, which provides the efficient volume description of an object. In engineering, the parametric solid modeling method is used for building B-Rep model. However, a geological modeling generally adopts discrete modeling based on the triangulated surface, called a Sealed Geological Model, which defines geological domain by using geological interfaces such as horizons, faults, intrusives and modeling boundaries. Discrete B-Rep model is incompatible with mesh generation softwares in engineering because of discrepancies between discrete and parametric technique. In this research we have developed a converting program from Sealed Geological Model to Gmsh and COMSOL software. The developed program can convert complex geological model built by geomodeling software to user-friendly FEM software and it can be applied to geoscience simulation such as geothermal, mechanical rock simulation etc.

The Terminal and Internal Hairpin Loops of the ctRNA of Plasmid pJB01 Play Critical Roles in Regulating Copy Number

  • Kim, Sam Woong;Jeong, In Sil;Jeong, Eun Ju;Tak, Je Il;Lee, John Hwa;Eo, Seong Kug;Kang, Ho Young;Bahk, Jeong Dong
    • Molecules and Cells
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    • v.26 no.1
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    • pp.26-33
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    • 2008
  • The plasmid pJB01, a member of the pMV158 family isolated from Enterococcus faecium JC1, contains three open reading frames, copA, repB, and repC. Plasmids included in this family produce counter-transcribed RNA (ctRNA) that contributes to copy number control. The pJB01 ctRNA, a transcript which consists of 54 nucleotides (nts), is encoded on the opposite strand from the copA/repB intergenic region and partially overlaps an atypical ribosome binding site (ARBS) for repB. The ARBS is integrated by the two underlined conserved regions: 5'-TTTTTGTNNNNTAANNNNNNNNNATG-3', and the ctRNA is complementary only to the 5' conserved sequence 5'-TTTTTGT-3'. This complementary sequence is located at a distance from the terminal loop of the ctRNA secondary structure. The ctRNA structure predicted by the mfold program suggests the possible generation of a terminal and an internal hairpin loop. The amount of in vitro translation product of repB mRNA was inversely proportional to the ctRNA concentration. Mutations in the terminal and internal hairpin loops of the ctRNA had inhibitory effects on its binding to the target mRNA. We propose that the intact structures of the terminal and internal hairpin loops, respectively, play important roles in forming the initial kissing and extending complexes between the ctRNA and target mRNA and that these regulate the copy number of this plasmid.

A Small Cryptic Plasmid pZMO1 of Zymomonas mobilis ATCC10988

  • Kang, Hyung-Lyun;Kang, Hyen-Sam
    • Genomics & Informatics
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    • v.1 no.1
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    • pp.55-60
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    • 2003
  • The nucleotide sequence of pZMO1, a small cryptic plasmid of Zymomonas mobilis ATCC10988 was determined. Analysis of 1,680 bp of sequence revealed $69\%$ identity with Shigella sonnei plasmid, pKYM and $61\%$ identity with Nostoc sp. ss DNA replicating plasmid. Analysis of a deduced amino acid sequence of an orf of pZMO1 revealed $75\%$ identity and $90\%$ similarity with the repA gene of Synechocystis sp. plasmid pCA2.4. The upstream region of the repA gene of pZMO1 possesses six directed repeat sequences and two inverted repeat sequences at downstream of the IR consensus sequence of nick region of rolling circle replication (RCR) plasmid. A typical terminator hairpin structure was found at the downstream region of repA gene. Degradation of single-stranded plasmid DNA by S1 nuclease was detected by Southern hybridization. It suggests that pZMO1 replicates by a rolling circle mechanism in Z. mobilis ATCC10988 cells.

Cloning and Base Sequence Determination of Replication Initiation Gene (rep) Isolated from Staphylococcus aureus DH1 R-plasmid pSBK203 (Staphylococcus aureus DH1에서 분리된 R-plasmid pSBK203의 복제 개시 유전자(rep) 분리 및 염기서열 결정)

  • Park, Seung-Moon;Kwon, Dong-Hyun;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.44-47
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    • 1993
  • A replication initiation gene was identified and its nucleotide sequence has been determined from a 3.8 kb, chloramphenicol acethyltransferase conferring R-plasmid pSBK203 of Staphylococcus aures. Location of the replication related region of pSBK 203 was determined by interuption with pUC 119 at XBaI and MspI sites which resulted in inactivation of replication in Bacilius subtilis. Base sequence of this region revealed on open reading frame of 942 base pairs, which encoded a 314 amino acid protein. Base sequence homology with other rep of pT181 family plasmids such as pT181, pC221, pC223, pS194, pU112, and pCW7 was ranged from 78% to 97% and the predicted amino acid sequence homology was from 72% to 95%.

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A Performance Comparison of 3-D Spatial Operation between Prism Model and B-Rep Model (프리즘 모델과 B-Rep 모델의 3차원 공간 연산 성능 비교)

  • Jang, Dae-Sung;Kim, Ho-Chul;Kim, Ji-Hyun;Kim, Joon-Seok;Li, Ki-Joune
    • Proceedings of the Korean Association of Geographic Inforamtion Studies Conference
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    • 2010.06a
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    • pp.303-305
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    • 2010
  • 실내공간의 특성을 고려하여 u-GIS 서비스를 제공하기 위해서는 그에 맞는 적절한 모델과 질의처리 방법이 필요하다. 프리즘 모델은 실내공간을 표현하기 위한 3차원 기하 모델이며, 이에 대한 질의처리 방법이 제안된 바 있다. 하지만 이에 대한 실험적 근거가 부족하므로, 본 논문에서는 프리즘 모델의 질의 처리 방법을 적용한 시스템과 B-Rep 기반의 3차원 공간 DBMS 시스템간의 연산 수행을 비교 실험하였다. 그 결과 프리즘 모델 기반의 질의처리 방법이 B-Rep 기반의 공간 DBMS 보다 질의 처리 성능이 우수한 것으로 검증되었다.

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