• Title/Summary/Keyword: 클론성

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Cloning of Coat Protein Gene from Korean Isolate Potato Leafroll Virus (PLRV) and Introduction into Potato (Solanum tuberosum) (한국 분리주 감자 잎말림 바이러스 (PLRV) 외피 단백질 유전자의 클로닝 및 감자 내 도입)

  • Seo Hyo-Won;Yi Jung-Yoon;Park Young-Eun;Cho Ji-Hong;Hahm Young-Il;Cho Hyun-Mook
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.243-250
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    • 2005
  • The coat protein gene (AF296280) of the Korean isolate Potato leafroll virus (PLRV) was cloned and the open reading frame (627 bp) was transformed into potato (Solanum tuberosum cv. Superior). Out of seventeen individual transgenic lines, five lines were identified to confer resistance to PLRV through the five generation's selection program in the greenhouse as well as isolated trial field. Successful introduction and genetic stability of coat protein gene in the genome of potato were confirmed by polymerase chain reaction (PCR), Southern blot hybridization and northern blot hybridization. Some of the transgenic lines were highly resistant to PLRV but did not show any resistance to less homologous Potato virus Y (PVY). Our results suggest that the resistance to PLRV is due to homology dependent gene silencing by sense strand coat protein gene. In addition, the results of field test through five generations showed that there were no significant differences comparing to nontransgenic potatoes in the morphological aspect of shoot as well as tuber, Ho remarkable differences were also observed in the major agronomic characters and yields except for the resistance to PLRV.

Development of Polyclonal Antibodies to Abdominal and Subcutaneous Adipocyte for Producing Fat-reduced High Quality Pork (저지방 고품질 돈육 생산을 위한 돼지 복강 및 피하지방 항체 개발)

  • Choi, Chang-Weon;Baek, Kyung-Hoon;Choi, Chang-Bon;Oh, Young-Kyoon;Hong, Seong-Koo
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.87-94
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    • 2010
  • The aim of the present study was to develop polyclonal antibodies to regional inedible adipocytes of pigs and investigate the effect of these antibodies on adipocytes in vitro. As antigens, abdominal and subcutaneous adipocyte PMPs from pigs were injected into sheep 3 times per 3 wk intervals for passive immunization, and non-immunized serum, antisera against abodominal (AAb) or subcutaneous adipocyte PMPs (SAb) were collected before and after the injections. Titers of the antisera obtained from sheep and their cross-reactivities with the heart, kidney, liver, lung, muscle, and spleen of pig were determined by ELISA. Isolation and culture of abdominal and subcutaneous adipocytes from pigs were performed to analyze LDH concentration. At a 1:1,000 dilution, little antibody reactivity was observed for non-immunized serum whereas both AAb and SAb had relatively strong reactivity up to a dilution of 1:16,000. These findings may indicate that strong antibodies against adipocyte PMPs can be developed using an immunological approach. Extremely low reactivity of AAb and SAb was detected with the PMPs of the organs. Both antisera most strongly reacted with each adipocyte PMPs and showed statistically (p<0.05) higher cross-reactivities compared with the non-immunized serum. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte PMPs are well developed and are safe against cross-reactivities with the organs of pigs. Further studies on the in vivo nutritional safety and fat reduction of these antibodies in pigs will be required fat-reduced high quality pork production.

Construction of the Genomic Expression Library of Bacillus anthracis for the Immunomic Analysis (면역체 분석을 위한 탄저균 유전자 발현 라이브러리의 구축)

  • Park, Moon-Kyoo;Jung, Kyoung-Hwa;Kim, Yeon-Hee;Rhie, Gi-Eun;Chai, Young-Gyu;Yoon, Jang-W.
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.21-26
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    • 2010
  • As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

Differential Intracellular Localization of Mitotic Centromere-associated Kinesin (MCAK) During Cell Cycle Progression in Human Jurkat T Cells (인체 Jurkat T 세포에 있어서 세포주기에 따른 MCAK 단백질의 세포 내 위치변화)

  • Jun Do Youn;Rue Seok Woo;Kim Su-Jung;Kim Young Ho
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.253-260
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    • 2005
  • Mitotic centromere-associated kinesin (MCAK), which is a member of the Kin I (internal motor domain) subfamily of kinesin-related proteins, is known to play a role in mitotic segregation of chromosome during M phase of the cell cycle. In the present study, we have produced a rat polyclonal antibody using human MCAK (HsMCAK) expressed in E. coli as the antigen. The antibody specifically recognized the HsMCAK protein (81 kDa), and could detect its nuclear localization in human Jurkat T cells and 293T cells by Western blot analysis. The specific stage of the cell cycle was obtained through blocking by either hydroxyl urea or nocodazole and subsequent releasing from each blocking for 2, 4, and 7 h. While the protein level of HsMCAK reached a maximum level in the S phase with slight decline in the $G_{2}-M$ phase, the electrophoretic mobility shift from $p81^{MCAK}\;to\;p84^{MCAK}$ began to be induced in the late S phase and reached a maximum level in the $G_{2}/M $ phase, and then it disappeared as the cells enter into the $G_{1}$ phase. Immunocytochemical analysis revealed that HsMCAK protein localized to centrosome and nucleus at the interphase, whereas it appeared to localize to the spindle pole, centromere of the condensed mitotic DNA, spindle fiber, or midbody, depending on the specific stage of the M phase. These results demonstrate that a rat polyclonal antibody raised against recombinant HsMCAK expressed in E. coli specifically detects human MCAK, and indicate that the electrophoretic mobility shift from $p81^{MCAK}\;to\;p84^{MCAK}$, which may be associated with its differential intracellular localization during the cell cycle, fluctuates with a maximum level of the shift at the $G_{2}-M$ phase.

A Study on the Application of Physical Soil Washing Technology at Lead-contaminated Shooting Range in a Closed Military Shooting Range Area (폐 공용화기사격장 내 납오염 사격장 군부지의 물리적 토양세척정화기술 적용성 연구)

  • Jung, Jaeyun;Jang, Yunyoung
    • Journal of Environmental Impact Assessment
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    • v.28 no.5
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    • pp.492-506
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    • 2019
  • Heavy metal contaminants in the shooting range are mostly present in a warhead circle or a metal fragment present as a particle, these fine metal particles are weathered for a long period of time is very likely that the surface is present as an oxide or carbon oxide. In particular, lead which is a representative contaminant in the shooting range soil, is present as more fine particles because it increases the softness and is stretched well. Therefore, by physical washing experiment, we conducted a degree analysis, concentration of heavy metals by cubic diameter, composition analysis of metallic substances, and assessment of applicability of gravity, magnetism and floating selection. The experimental results FESEM analysis and the measurement results lead to the micro-balance was confirmed thatthe weight goes outless than the soil ofthe same size in a thinly sliced and side-shaped structure according to the dull characteristics it was confirmed that the high specific gravity applicability. In addition, the remediation efficiency evaluation results using a hydrocyclone applied to this showed a cumulative remediation efficiency of 71%,twice 80%, 3 times 91%. On the other hand, magnetic sifting showed a low efficiency of 17%,floating selection -35mesh (0.5mm)target soil showed a relatively high efficiency to 39% -10mesh (2mm) efficiency was only 16%. The target treatment diameter of soil washing should be 2mm to 0.075mm, which is applied to the actual equipment by adding an additional input classification, which would require management as additional installation costs and processes are constructed. As a result, it is found that the soilremediation of shooting range can be separately according to the size of the warhead. The size is larger than the gravel diameter to most 5.56mm, so it is possible to select a specific gravity using a high gravity. However, the contaminants present in the metal fragments were found to be processed by separating using a hydrocyclone of the soil washing according to the weight is less than the soil of the same particle size in a thinly fragmented structure.

Imaging for Multiple Myeloma according to the Recent International Myeloma Working Group Guidelines: Analysis of Image Acquisition Techniques and Response Evaluation in Whole-Body MRI according to MY-RADS (International Myeloma Working Group의 최신 가이드 라인에 따른 다발성 골수종의 영상검사법 및 MY-RADS에 따른 전신 MRI에서의 영상 획득과 반응 평가 소개)

  • A Yeon Son;Hye Won Chung
    • Journal of the Korean Society of Radiology
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    • v.84 no.1
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    • pp.150-169
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    • 2023
  • Multiple myeloma (MM) is a malignant hematologic disease caused by the proliferation of clonal plasma cells in the bone marrow, and its incidence is increasing in Korea. With the development of treatments for MM, the need for early diagnosis and treatment has emerged. In recent years, the International Myeloma Working Group (IMWG) has been constantly revising the laboratory and radiological diagnostic criteria for MM. In addition, as whole-body MRI (WBMR) has been increasing used in the diagnosis and treatment response evaluation of patients with MM, the Myeloma Response Assessment and Diagnosis System (MY-RADS) was created to standardize WBMR image acquisition techniques, image interpretation, and response evaluation methods. Radiologists need to have a detailed knowledge of the features of MM for accurate diagnosis. Thus, in this review article, we describe the imaging method for MM according to the latest IMWG guidelines as well as the image acquisition and response evaluation technique for WBMR according to MY-RADS.

Molecular Cloning and Expression of the Novel Attacin-Like Antibacterial Protein Gene Isolated from the Bombyx mori (집누에로부터 새로운 attactin 유산 항세균성 펩타이드 유전자의 분리 및 발현)

  • 윤은영;김상현;강석우;진병래;김근영;김호락;한명세;강석권
    • Korean journal of applied entomology
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    • v.36 no.4
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    • pp.331-340
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    • 1997
  • Hyalophora cecropia attacin-like antibacterial gene was isolated from Bombyx mori induced with nonpathogenic bacteria. It was expressed in Spodopfera frugiperda 9 (Sf9 cells using baculovirus expression vector system (BEVS), and examined its antibacterial activity. With a cDNA library constructed from fifthinstar B. mori injected with Escherichia coli(4 X IOhcellsllarva), differential screening was performed using naive and induced mRNA probes. BmInc6 clone was screened by partial nucleotide sequence and GenBank database analysis. A complete nucleotide sequence of Bmlnc6 cDNA was determined (GenBank, AF005384). Its insert size was 852 bp and had open reading frame that started translation at position 35 and stopped at 679. And its putative polyadenylational signal existed at 812 bp. The number of amino acid deduced from Bmlnc6 cDNA was 214 and hydropathy analysis showed that this peptide was hydrophilic. This peptide deduced by BmInc6 was named nuecin. When the nuecin gene was expressed in Sf9 cells using BEVS, about 950 bp of the transcripts was detected. In addition, SDS-PAGE analysis showed that the molecular weights of intracellular expressed protein and the mature protein secreted to culture media were approximately 23 and 20 kDa, respectively. The antibacterial activity of nuecin against E. coli and Bacillus subtilis was significantly high, demonstrating that nuecin had a wider antibacterial spectrum with gram negative and positive bacteria than attacin.

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Fact-finding Survey on Occurrence of Weeds and Herbicide Usage for Paddy Rice Cultivation in Gyeonggi Province, Korea (경기지역 논잡초 발생양상 및 제초제 사용실태)

  • Won, Taejin;Park, Jungsu;Kim, Soonjae;Kim, Heedong
    • Weed & Turfgrass Science
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    • v.2 no.4
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    • pp.352-357
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    • 2013
  • A survey was performed to investigate major paddy field weeds and frequently used herbicides in Gyeonggi province, Korea in order to establish a system to efficiently control herbicide-resistant weeds. The dominant paddy field weeds included Echinochloa spp. (22%), Eleocharis kuroguwai (14%), Sagittaria trifolia (13%), Scirpus juncoides (12%), Monochoria vaginalis (9%) and Sagittaria pygmaea (7%), and the most widely used herbicides were butachlor 33% CS (25%), benzobicyclon + fentrazamide + imazosulfuron 11.5% SC (9%), cyclosulfamuron + mefenacet 22.2% SC (9%), butachlor 5% GR (6%), and oxadiazon 12% EC (6%) in Gyeonggi province, Korea. Many paddy rice growers considered S. juncoides, M. vaginalis, S. pygmaea and C. difformis as herbicide-resistant weeds. An extensive research deserves to be conducted to monitor occurrence of herbicideresistant paddy weeds in Gyeonggi province, Korea.

Antigenic Determinant Mapping in preS2 Region of Hepatitis B Surface Antigen (B형 간염바이러스 표면항원 preS2 부위의 항원결정인자 규명)

  • 권기선;김창수;박주상;한문희;유명희
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.13-18
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    • 1990
  • A DNA sequence encoding the adr subtype preS2 region of hepatitis B virus envelope protein was fused to 5' end of lacZ gene yielding a plasmid pTSZ, in order to produce a preS2-$\beta$-galactosidase fusion protein. Serial deletions from 3' and 5' end of preS2 were constructed in plasmids, which were expressed and their antigenicities were examined with the monoclonal antibody H8. Deletions from amino and carboxy terminal to certain points did not affect the antigenicity, but the longer deletions destroyed the antigenicity. End points of deleted preS2 sequence were determined by DNA sequencing. As a result, each end of preS2 epitope was located in the region of amino acid residue 130-132 and 140-142, respectively. Residue 143 may be supplementary for antigenic epitope since the deletion from carboxy terminal to residue 143 revealed partial defect of antigenicity. In the interval of antigenic epitope the amino acid differences between adr and adw2 subtype occurred ar residue 130, 132, and 141. This result indicated that one or more of the three residues are responsible for the binding specificity of monoclonal antibody H8 to adr subtype preS2 fusion protein.

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Potential of Contaminant Removal Using a Full-Scale Municipal Water Treatment System with Adsorption as Post-Treatment (실 규모 물 처리 공정 및 후속 흡착 처리에 의한 오염원 제거 잠재성 평가)

  • Haeil Byeon;Geonhee Yeo;Anh-Hong Nguyen;Youngwoong Kim;Donggun Kim;Taehun Lee;Seolhwa Jeong;Younghoa Choi;Seungdae Oh
    • Land and Housing Review
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    • v.15 no.1
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    • pp.167-177
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    • 2024
  • This study aimed to assess the efficacy of an adsorption process in removing organic matter and micropollutant residuals. After a full-scale water circulation system, the adsorption process was considered a post-treatment step. The system, treating anthropogenically impacted surface waters, comprises a hydro-cyclone, coagulation, flocculation, and dissolved air flotation unit. While the system generally maintained stable and satisfactory effluent quality standards over months, it did not meet the highest standard for organic matter (as determined by chemical oxygen demands). Adsorption experiments utilized two granular activated carbon types, GAC 830 and GCN 830, derived from coal and coconut-shell feedstocks, respectively. The assessment encompassed organic materials along with two notable micropollutants: acetaminophen (APAP) and acid orange 7 (AO7). Adsorption kinetics and isotherm experiments were conducted to determine adsorption rates and maximum adsorption amounts. The quantitative findings derived from pseudo-second-order kinetics and Langmuir isotherm models suggest the effectiveness of the adsorption process. The findings of this study propose the potential of employing the adsorption process as a post-treatment to enhance the treatment of contaminants that are not satisfactorily treated by conventional water circulation systems. This enhancement is crucial for ensuring the sustainability of urban water cycles.