• Title/Summary/Keyword: integration site

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Site-Specific Recombination by the Integrase MJ1 on Mammalian Cell (동물 세포 내에서 MJ1 인티그라제에 의한 부위 특이적 재조합)

  • Kim, Hye-Young;Yoon, Bo-Hyun;Chang, Hyo-Ihl
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.337-344
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    • 2011
  • Integrase MJ1 from the bacteriophage ${\Phi}FC1$ carries out recombination between two DNA sequences (the phage attachment site, attP and the bacterial attachment site, attB) in NIH3T3 mouse cells. In this study, the integration vector containing attP, attB and the integrase gene MJ, was constructed. The integration mediated by integrase MJ1 in Escherichia coli led to excision of LacZ. Therefore, the frequency of integration was measured by the counting of the white colony, which is detectable on X-Gal plates. The extrachromosomal integration in NIH3T3 mouse cells was monitored by the expression of the green fluorescent protein (GFP) as a reporter. To demonstrate integration mediated integrase MJ1 in NIH3T3 cells, vectors containing attP and attB were co-transfected into NIH3T3 cells. The integration was confirmed by fluorescent microscopy. The expression of GFP was induced in NIH3T3 cells expressing MJ1 without accessory factors. By contrast, the excision mediated by the MJ1 between attR and attL had no effect on the expression of GFP. These results suggest that integrase MJ1 may enable a variety of genomic modifications for research and therapeutic purposes in higher living cells.

Biochemical Properties of Second Site Mutation of Human Immunodeficiency Virus Integrase

  • Kim, Do-Jin;Oh, You-Take;Lee, Sang-Kwang;Shin, Cha-Gyun
    • BMB Reports
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    • v.32 no.6
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    • pp.599-604
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    • 1999
  • A highly conserved amino acid, glutamic acid (Glu), present at position 152 in the catalytic domain of the human immunodeficiency virus type 1 (HIV-1) integrase (IN) protein has been known to be critical for enzymatic function since substitution of Glu 152 with other residues results in a complete loss of enzymatic activities. In order to better understand the role of Glu 152 as a conserved residue in enzymatic action, intragenic second site mutations have been introduced around residue 152 of a mutant IN (E152A), and their biochemical properties were analyzed in terms of enzymatic activities. Disintegration activities were found to be significantly restored in several second site mutant INs, while integration activities were only recovered weakly. However, endonucleolytic activities were not discovered in all the mutant INs. These findings indicate that the second site mutations can partially restore that catalytic structure of the active site disturbed by the E152A mutation and lead to the regaining of integration and disintegration activities. In addition, it is also suggested that endonucleolytic activity requires a more accurate structure of the catalytic site than that for the integration and disintegration activities.

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Development of Site Classification System and Modification of Design Response Spectra considering Geotechnical Site Characteristics in Korea (III) - Modification of Desing Response Specra (국내 지반특성에 적합한 지반분류 방법 및 설계응답스펙트럼 개선에 대한 연구 (III) - 설계응답스펙트럼 개선방법)

  • Yoon, Jong-Ku;Kim, Dong-Soo;Bang, Eun-Seok
    • Journal of the Earthquake Engineering Society of Korea
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    • v.10 no.2 s.48
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    • pp.63-71
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    • 2006
  • In the companion paper (II, Development of Site Classification System), new size classification system based on site periods $(T_G)$ was recommended for legions of shallow bedrock depth in Korea. Despite the site classification method was improved, the response spectrum would be required to be modified by adjusting the integration interval to calculate the site coefficients because the response spectra did not match well the average spectral accelerations obtained by site response analyses in the range of long periods. In this paper, new response spectra for each site categories were determined by adjusting the integration interval of long period site coefficient $F_v\;from\;0.4{\sim}2.0\;to\;0.4{\sim}1.5$ second. It matched well the average spectral accelerations and new response spectrum, and it was also improved compared to the current she classification system.

Modification of Design Response Spectra Considering Geotechnical Site Characteristics in Korea (국내 지반특성에 적합한 설계응답스펙트럼 개선을 위한 증폭계수 재산정에 대한 연구)

  • Yoon, Jong-Ku;Kim, Dong-Soo;Bang, Eun-Seok
    • Proceedings of the Earthquake Engineering Society of Korea Conference
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    • 2006.03a
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    • pp.113-124
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    • 2006
  • Despite the site classification method was improved in the previous study, the response spectrum would be required to be modified by adjusting the integration interval to calculate the site coefficients because the response spectra did not match well the average spectral accelerations obtained by site response analyses in the range of long periods. In this paper, new response spectra for each site categories were determined by adjusting the integration interval of long-period site coefficient $F_{v}$ from $0.4{\sim}2.0$ to $0.4{\sim}1.5$ second. It matched well the average spectral accelerations and new response spectrum, and it was also improved compared to the current site classification system.

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Development of a Food-Grade Integration Vector for Heterologous Gene Expression and Protein Secretion in Lactococcus lactis

  • Jeong, Do-Won;Lee, Jong-Hoon;Kim, Kyoung-Heon;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1799-1808
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    • 2006
  • A food-grade integration vector based on site-specific recombination was constructed. The 5.7-kb vector, pIMA20, contained an integrase gene and a phage attachment site originating from bacteriophage A2, with the ${\alpha}$-galactosidase gene from Lactobacillus plantarum KCTC 3104 as a selection marker. pIMA20 was also equipped with a controllable promoter of nisA ($P_{nisA}$) and a signal peptide-encoding sequence of usp45 ($SP_{usp45}$) for the production and secretion of foreign proteins. pIMA20 and its derivatives mediated site-specific integration into the attB-like site on the Lactococcus lactis NZ9800 chromosome. The vector-integrated recombinant lactococci were easily detected by the appearance of blue colonies on a medium containing $X-{\alpha}-gal$ and also by their ability to grow on a medium containing melibiose as the sole carbon source. Recombinant lactococci maintained these traits in the absence of selection pressure during 100 generations. The ${\alpha}-amylase$ gene from Bacillus licheniformis, lacking a signal peptide-encoding. sequence, was inserted downstream of $P_{nisA}\;and\;SP_{usp45}$ in pIMA20, and the plasmid was integrated into the L. lactis chromosome. ${\alpha}-Amylase$ was successfully produced and secreted by the recombinant L. lactis, controlled by the addition and concentration of nisin.

Automated 3D Model Reconstruction of Disaster Site Using Aerial Imagery Acquired By Drones

  • Kim, Changyoon;Moon, Hyounseok;Lee, Woosik
    • International conference on construction engineering and project management
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    • 2015.10a
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    • pp.671-672
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    • 2015
  • Due to harsh conditions of disaster areas, understanding of current feature of collapsed buildings, terrain, and other infrastructures is critical issue for disaster managers. However, because of difficulties in acquiring the geographical information of the disaster site such as large disaster site and limited capability of rescue workers, comprehensive site investigation of current location of survivors buried under the remains of the building is not an easy task for disaster managers. To overcome these circumstances of disaster site, this study makes use of an unmanned aerial vehicle, commonly known as a drone to effectively acquire current image data from the large disaster areas. The framework of 3D model reconstruction of disaster site using aerial imagery acquired by drones was also presented. The proposed methodology is expected to assist rescue workers and disaster managers in achieving a rapid and accurate identification of survivors under the collapsed building.

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Molecular Characterization of the Region Encoding Integrative Functions from Enterococcal Bacteriophage ${\phi}$FC1

  • Kim, Min-Jung;Lee, Jin-Young;Kim, Young-Woo;Sung, Ha-Chin;Chang, Hyo-Ihl
    • BMB Reports
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    • v.29 no.5
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    • pp.448-454
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    • 1996
  • Bacteriophage ${\phi}FC1$ is a temperate phage which was identified as a prophage in the Enterococcus faecalis KBL703 chromosome. Phage ${\phi}FC1$ integrates into the host chromosome by site-specific recombination. The phage attachment site P (attP) was localized within the 0.65-kb XhoI-HindIII fragment and the nucleotide sequence of the region was determined. An open reading frame (mj1) which adjoined the phage attachment site encoded a deduced protein related to the site-specific recombinase family. The organization of this region was comparable to other site-specific recombination systems. The molecular weight of the expressed MJ1 in E. coli was in good agreement with the predicted 53,537 Da of the mj1 gene product. Elucidation of the phage-specific integration process in this study would provide useful genetic tools such as a chromosomal integration system.

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The Development for KASS Reference Station Site (KASS 기준국 사이트 구축)

  • Cho, Sunglyong;Jang, Hyunjin;Jeong, Hwanho;Lee, Byungseok;Nam, Giwook
    • Journal of Advanced Navigation Technology
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    • v.24 no.4
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    • pp.273-279
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    • 2020
  • In the Korea's SBAS(KASS), reference site is an important infrastructure facility for the collecting and monitoring GPS/GEO signals. The SBAS reference station has an clear requirements than other regular monitoring stations. It requires constant maintenance during the system operation. The development for KRS site should be prepared for site survey, site construction, antenna geodetic survey, equipment installation and operation. Site survey is initially performed as an important step to predict site availability and system performance. The operation center must provide the reference site, equipment room, and appurtenant to satisfy the site requirements. The position of antennas is very important information, and accuracy must be secured through the geodetic survey. Measurement collected at the from precise antenna are provided to the KASS processing station. The position of antenna should be maintained through continuous position checks and updates during the operation. When the development of the KRS site is completed, it performs tasks for installing and operating the KRS equipment. In this paper, we presented the procedures and some results for the development of the 7 KRS sites.

Constrution and Application of Underground Facilities Survey System using the 3D Integration Map of Underground Geospatial Information (3차원 지하공간통합지도를 활용한 지하시설물 현장 측량 시스템 구축 및 적용)

  • SONG, Seok-Jin;CHO, Hae-Yong;HEO, Hyun-Min;KIM, Sung-Gil
    • Journal of the Korean Association of Geographic Information Studies
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    • v.24 no.4
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    • pp.164-173
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    • 2021
  • Recently, as underground space safety issues such as sink hole, ground subsidence and damage to old underground facilities have been increasing in urban areas, the precise management of underground facilities ins more required. Thus, this study developed a function to that, visualize on Integration Map of Underground Geospatial Information a real-time survey data of underground facilities acquired on site or underground facility survey data acquired through on-site survey after underground facility exploration and developed a function convert to surveying-results. In addition, using the on-site survey performance utilization function in connection with the Integration Map of Underground Geospatial Information developed through this study, the surveying -results obtained with the Total-station at the water pipeline burial construction site in Eunpyeong-gu, Seoul are visualized on the Integration Map of Underground Geospatial Information and On-site verification was performed by converting spatial-information performance files and transmitting the Integration Map of Underground Geospatial Information to the mobile center. Based on this, it was possible to verify the work procedure using the surveying-results in the area where the Integration Map of Underground Geospatial Information was built, and to review the direction of future improvement directions.

Study of Selection Plan of Circuit breakers, Cables and Modeling of Korean Low Voltage Electrical Installation integration Test Site based on IEC 60364 (IEC 60364 기반의 한국형 저압전기설비 통합 실증단지 모델링 및 차단기와 케이블의 선정 방안 고찰)

  • Kim, Doo-Ung;Ryu, Kyu-Sang;Kim, Han-Soo;Shin, Dae-Sung;Ryu, Ki-Hwan;Kim, Chul-Hwan
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.29 no.9
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    • pp.59-64
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    • 2015
  • IEC is an international standards which are used in many countries with Europe as the center. IEC standard is introduced in Korea according to WTO/TBT agreements, however until now there are no buildings in Korea which are designed applying IEC standard. Therefore, KEA(Korea Electric Association) is scheduled to construct Korean low voltage electrical installation integration test site which is designed applying IEC standard. In this paper, before being under construction of Korean low voltage electrical installation integration test site, power substation is modeled based on real design parameters and method to select circuit breakers and cables is presented applying IEC standard in the modeled power substation. EMTP(ElctroMagnetic Transient Program) is used for simulation program. EMTP which is power system analysis program is easy to model power system and power substation.