• Title/Summary/Keyword: 중합효소 연쇄반응

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A Case of T-cell Primary Pulmonary Lymphoma Diagnosed by $TCR{\gamma}$ gene rearrangement ($TCR{\gamma}$ 유전자 재배열로 진단된 T세포 원발성 폐림프종 1예)

  • Choi, Young Mee;Kim, Seung Joan;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyoung;Moon, Hwa Sik;Park, Sung Hak;Song, Jeong Sup
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.1001-1007
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    • 1996
  • 반복되는 흉막염 및 폐렴을 주소로 내원한 30세 남자환자에서 경기관지 폐생검과 흉수내 림프구에 대한 유세포 분석 및 T세포 수용체 유전자 재배열 분석을 실시하였다. 경기관지 폐생검 조직의 연역조직화학 염색상 대부분의 림프구가 T세포 표식자인 UCHLl 에 대해 강하게 염색되었고, B세포 표식자인 L26에 대해서는 거의 염색되지 않았다. 흉수에서 추출한 림프구의 유세포 분석상 CD3양성 CD2양성인 T림프구가 대부분이었고, 이들 림프구에 대해 중합효소연쇄반응을 이용한 T세포 수용체 유전자 재배열 분석을 하였더니 $TCR{\gamma}$ 유전자 재배열과 클론성을 관찰할 수 있었다. T세포 원발성 폐림프종으로 진단하였고, 문헌고찰과 함께 보고하는 바이다.

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The Detection of Genetically Modified Organisms in Soybean by DHPLC and Polymerase Chain Reaction (DHPLC와 중합효소연쇄반응에 의한 유전자재조합 콩의 검출)

  • Lee, Kyoung-Hae;Park, Su-Min
    • Food Science and Preservation
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    • v.15 no.1
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    • pp.88-93
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    • 2008
  • This paper focused on the detection of the genetically modified soybean (Glycine max L. MERRILL) samples to search for the speedy analysis methods. We have identified the PCR (polymerase chain reaction) assay with a newly developed technique called DHPLC (denaturing high performance liquid chromatography) to screen the GMO in soybean. The DHPLC is i1s ability to directly detection specific sequences of DNA by using column. With these characteristics. the DHPLC assay had the advantage of simplicity, rapidty could obtain result within 20 minutes. Whereas $15{\times}10^{-4}ng/{\mu}L$ concentration could be detected with the PCR analysis, $15{\times}10^{-5}ng/{\mu}L$ concentration could be detected with the DHPLC method. Therefore, DHPLC method was considered to be a simple, fast and sensitivity screening method rather than PCR analysis for GMO detection in soybean.

Detection of Novel Polyketide Synthase Genes in Sorangium cellulosum Isolated in Korea (국내에서 분리한 Sorangium cellulosum의 신규 Polyketide Synthase 유전자 검출)

  • Youn, Jin-Kwon;Kim, Do-Hee;Lee, Han-Bit;Lee, Kye-Won;Cho, Kyung-Yun
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.136-143
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    • 2010
  • DNA fragments encoding the ketosynthase (KS) domain of polyketide synthase (PKS) genes were amplified using polymerase chain reaction (PCR) from 9 strains of Sorangium cellulosum isolated in Korea, cloned into a plasmid vector and sequenced. A total of 83 cloned DNA fragments were analyzed, and similar fragments were excluded, leaving 43 independent DNA fragments encoding the KS domains. The predicted amino acid sequences of 32 fragments were 70%-100% identical to the amino acid sequences of already known PKS genes, while the remaining 11 fragments were $\leq$67% or less identical to the known sequences, suggesting that these genes are novel PKS genes.

Mutation Analysis in β2-Adrenergic Receptor Gene by Single Strand Conformation Polymorphism (SSCP) and Denaturing High Performance Liquid Chromatography (DHPLC) (SSCP와 DHPLC에 의한 β2-교감신경수용체 유전자의 돌연변이 분석)

  • Park, Sang-Bum;Han, Sang-Man;Nam, Youn-Hyoung;Jang, Won-Cheoul
    • Analytical Science and Technology
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    • v.17 no.1
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    • pp.53-59
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    • 2004
  • Up to now, methods for the detection of genetic alterations as single strand conformation polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE) have been used. It is too labor-intensive and expensive to serve for routine analysis. Moreover, lower in its sensitivity and specificity being also strongly dependent on the experience of the investigater. To improve these problems, we analysed mutation of ${\beta}_2$-adrenergic receptor gene that controls bronchial asthma by denaturing high performance liquid chromatography (DHPLC) according to ion-pair reversed phase chromatography (IP-RPC). We extracted genomic DNA from 80 asthma patients and then amplified DNA using PCR and analysed PCR product by SSCP and DHPLC. As a result, we analysed mutation frequency is 19 (23.75%) on SSCP and 25 (31.25%) on DHPLC in ${\beta}_2$-adrenergic receptor gene. We conclude that DHPLC is a fast and simple screening method rather than SSCP analysis.

Evaluation of the preservation state of human skeletal remains using real-time PCR (출토 인골 DNA의 real-time PCR 정량에 의한 보존상태 평가 연구 - 부여 오수리 출토 인골을 중심으로 -)

  • Kwon, Eun-Sil;Cho, Eun-Min;Kim, Sue-Hoon;Kang, Soyeong
    • 보존과학연구
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    • s.32
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    • pp.171-183
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    • 2011
  • In this study molecular genetic analysis was carried out on 4 human skeletal remains from Osuri, Buyeo. We showed that real-time PCR is the method of the choice to assess the initial number of genuine ancient DNA molecules. Human mitochondrial DNA quantification was accomplished by the real-time PCR for the cytochrome b gene of the mitochondria. Histological results proved to be a good potentiality for biochemical analysis using biomolecule. The level of specimen's preservation state was proved that level of quantitative result was BO-04, BO-01, BO-03, BO-02. Continually, we showed that biochemical and biomolecule results for the level of preservation state were similar. This study will be useful to important material for predicting biochemistry and biology analysis of the ancient bone.

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Identical small subunit ribosomal RNA gene nucleotide sequence of bovine Theileria isolates (Korea and Japan) and Theileria buffeli (Marula, Kenya) (한국파 일본의 소에서 분리한 Theileria 분리주와 Theiferia buffeli (Marula, Kenya)의 small subunit ribosomal RNA 유전자 염기서열의 일치)

  • 채준석;권오덕
    • Parasites, Hosts and Diseases
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    • v.36 no.1
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    • pp.47-54
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    • 1998
  • Small subunit ribosomal RNA (SSU rRNA) gene nucleotide sequences of bovine ReiLerin isolates from Korea (KLS and KCB) and japan (JHS) were determined. The genes from each isolate were amplified by the polymerase chain reaction and the approxi- mately 1.8 kb product cloned and sequenced by a modified dideoxynucleotide method. Overlapping gene segments produced with a series of primers were sequenced, resoRting in a complete DNA sequence for both forward and reverse strands of the SSU rRNA genes of each isolate. SSU rRNA gene sequences (termed Type A) were identical among the bovine ReiLeri,n isolates from Korea and the isolate from Japan. A GenBank data library homolo- gy search showed the sequence to be the same as that listed as leiLeyia buKeLi isolated from cattle in Marula, Kenya.

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Survey on the Occurrence of Five Grapevine Viruses in Korean Vineyards in 2021 (2021년 국내 포도원에서의 5종 포도 바이러스 감염 실태 조사)

  • Yeojin Choi;Hae-Ryun Kwak;Myung-Kyu Song;Mikyeong Kim
    • Research in Plant Disease
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    • v.30 no.2
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    • pp.176-180
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    • 2024
  • In this study, a total of 217 samples were collected from five grapevine cultivars in Chungcheongbuk-do and Gyeongsangbuk-do in 2021. These samples were tested for the presence of five grapevine viruses using specific primers through polymerase chain reaction. The results indicated that grapevine fleck virus was confirmed in 126 samples (58.06%), grapevine leafroll-associated virus (GLRaV)-3 in 112 samples (51.61%) and GLRaV-1 in four samples (1.84%). The infection rate was high in Shine Muscat and Alexandria cultivars, and by region, the Gyeongbuk region showed a high infection rate. However, the presence of grapevine red blotch associated virus and grapevine berry inner necrosis virus were not detected.

Anti-inflammatory Activity of Antimicrobial Peptide Zophobacin 1 Derived from the Zophobas atratus (아메리카왕거저리 유래 항균 펩타이드 조포바신 1의 항염증활성)

  • Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Seo, Minchul;Kim, Mi-Ae;Lee, Hwa Jeong;Baek, Minhee;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.30 no.9
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    • pp.804-812
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    • 2020
  • The giant mealworm beetle, Zophobas atratus (Coleoptera: Tenebrionidae) has been used as a protein source for small pets and mammals. Recently, it was temporarily registered in the list of the Food Code. We previously performed an in silico analysis of the Zophobas atratus transcriptome to identify putative antimicrobial peptides and identified several antimicrobial peptide candidates. Among them, we assessed the antimicrobial and anti-inflammatory activities of zophobacin 1 that was selected bio-informatically based on its physicochemical properties against microorganisms and mouse macrophage Raw264.7 cells. Zophobacin 1 showed antimicrobial activities against microorganisms without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that zophobacin 1 reduced expression levels of pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). We also investigated expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) production through quantitative real time-PCR and ELISA. Zophobacin 1 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) signaling. We confirmed that zophobacin 1 bound to bacterial cell membranes via a specific interaction with lipopolysaccharides. These data suggest that zophobacin 1 could be promising molecules for development as antimicrobial and anti-inflammatory therapeutic agents.

The Prevalence of Oral Spirochetes in Korean Adult Periodontitis (한국인 성인성 치주염 환자에서의 구강 스피로헤타의 분포)

  • Kim, Hay-Hyun;Choi, Bong-Kiu;Choi, Seong-Ho;Chai, Jung-Kiu;Kim, Chong-Kwan;Cho, Kyoo-Sung
    • Journal of Periodontal and Implant Science
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    • v.28 no.4
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    • pp.659-678
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    • 1998
  • In the present study, oligonucleotide probes based on 16S rRNA were taken to investigate the diversity of oral spirochetes without culture method. This is the first study that revealed oral spirochetes of both presently cultivable and uncultured oral spirochetes in Korean adult periodontitis patients. Subgingival plaque samples were taken from diseased sites(probing depth ${\geq}6\;mm$, experimental group, n=116) and healthy sites(probing depth${\leq}3mm$, control 1 group, n=28) in 29 patients with adult periodontitis, and from 20 periodontally healthy subjects(probing depth${\leq}3mm$, control 2 group, n=100). Following being examined under phase-contrast microscope, all samples were submitted to dot-blot hybridization after polymerase chain reacton with eubacterial primers. 5 species-specific probes(TVIN, TDEN, TMAL, TSOC, and TPEC) and 7 group-specific probes(TRE I, TRE II, TRE III, TRE IV, TRE V, TRE VI, and TRE VII) were used one by one for the identification of both cultivable and so far uncultivable oral spirochetes. All probes were labeled with digoxigenin(DIG)-ddUTP and detected by chemilumininescence. The following results were obtained. 1. Under phase-contrast microscope, 91.37% and 14.28% of oral spirochetes were observed in the experimental and control 1 groups, respectively. None of oral spirochetes were observed in control 2 group. 2. With universal probe, 98.27%, 46.42%, and 22.0% of oral spirochetes were observed in experimental, control 1, and control 2 groups, respectively. 3. With specific probe, 95.68%, 35.71%, and 19.0% of oral spirochetes were observed in experimental, control 1, and control 2 groups, respectively. 4. With species-specific probes, T. socranskii were recovered in a high percentage of sites(81.89%) examined, followed by T. maltophilum(50.0%), T. vincentii(36.20%), T. denticola(13.79%), respectively. With group- specific probes, TRE IV was recovered in a high percentage of sites(85.34%) examined, followed by TRE II(77.58%), TRE I(56.89%), TRE III(25.86%), TRE VI(5.17%), and TRE V(2.58%), respectively. 5. T. vincentii were only observed in the diseased sites, not in the healthy sites. 6. Neither T. pectinovorum nor group VII oral spirochetes were observed in any sites. The findings warrant further investgations of the recovered spirochetes to elucidate the possible associations of oral spirochetal prevalence in race and types of periodontitis, pathogenesis of T. vincentii and the possible distributional change of oral spirochetes before and after treatments.

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Establishment of a Radiation-Induced Fibrosis Model in BALB/c Mice (BALB/c 마우스를 이용한 방사선섬유증 모델 확립)

  • Ryu, Seung-Hee;Lee, Sang-Wook;Moon, Soo-Young;Oh, Jeong-Yoon;Yang, Youn-Joo;Park, Jin-Hong
    • Radiation Oncology Journal
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    • v.28 no.1
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    • pp.32-38
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    • 2010
  • Purpose: Although radiation-induced fibrosis is one of the common sequelae occurring after irradiation of skin and soft tissues, the treatment methods are not well standardized. This study aimed to establish the skin fibrosis mouse model by fractionated radiation for the further mechanism studies or testing the efficacy of therapeutic candidates. Materials and Methods: The right hind limbs of BALB/c mice received two fractions of 20 Gy using a therapeutic linear accelerator. Early skin damages were scored and tissue fibrosis was assessed by the measurement of a leg extension. Morphological changes were assessed by H&E staining and by Masson's Trichrome staining. TGF-${\beta}1$ expression from soft tissues was also detected by immunohistochemistry and PCR. Results: Two fractions of 20 Gy irradiation were demonstrated as being enough to induce early skin damage effects such as erythema, mild skin dryness, dry and wet desquamation within several weeks of radiation. After 13 weeks of irradiation, the average radiation-induced leg contraction was $11.1{\pm}6.2mm$. Morphologic changes in irradiated skin biopsies exhibited disorganized collagen and extracellular matrix fibers, as well as the accumulation of myofibroblasts compared to the non-irradiated skin. Moreover, TGF-${\beta}1$ expression in tissue was increased by radiation. Conclusion: These results show that two fractions of 20 Gy irradiation can induce skin fibrosis in BALB/c mice accompanied by other common characteristics of skin damages. This animal model can be a useful tool for studying skin fibrosis induced by radiation.