• Title/Summary/Keyword: PCR 동정

Search Result 458, Processing Time 0.028 seconds

Purification and Identification of Antioxidant Compounds from Dolichos lablab L. Seeds (백편두의 항산화 물질 분리 및 동정)

  • Kwon, Nam Woo;Kim, Jae Yeon;Cho, Yong Beom;Hwang, Bang Yeon;Kim, Jun Gu;Woo, Sun Hee;Lee, Moon Soon
    • Korean Journal of Medicinal Crop Science
    • /
    • v.27 no.6
    • /
    • pp.419-426
    • /
    • 2019
  • Background: This study aimed to identify antioxidant compounds from the seeds of Dolichos lablab L. by bioassay-guided isolation and recrystallization. Methods and Results: The water layer of D. lablab L. seed extract inhibits intracellular reactive oxygen species (ROS) expressing the 2',7'-dichlorofluorescein diacetate (DCF-DA), Cu/Zn superoxide dismutase (SOD) and catalase genes, as determined by quantitative real-time PCR (qRT-PCR). Two compounds were purified from the water layer of the seeds of D. lablab L. using column chromatography and prep-high performance liquid chromatography (HPLC). Using nuclear magnetic resonance (NMR) and electrospray Ionization mass spectrometry (ESI-MS), their chemical structures were identified as 5-[(2-acetyl-2,3-dihydro-1H-indazol-1-yl)carbonyl]-4,5-dihydro-3H-furan-2-one (C14H14N2O4) and stachyose. Conclusions: Two active antioxidant compounds were purified from the seed extract of D. lablab L. seed extract and the structures of these compounds were identified as C14H14O4N2 and stachyose.

THE ANTIBIOTIC ACTIVITY OF ACTINOMYCES ISOLATED FROM PLAQUE OF BLACK STAINED PRIMARY TEETH TO STREPTOCOCCUS MUTANS (Black stain을 가진 유치 치태에서 추출한 방선균의 S. mutans에 대한 항생능 평가)

  • Park, Soo-Jin;Kim, Shin;Jeong, Tae-Sung;Kim, Jae-Moon
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.36 no.1
    • /
    • pp.12-19
    • /
    • 2009
  • The aim of this study is to assess the antibiotic activity of Actinomyces in plaque from black stained primary teeth to Streptococcus mutans. Samples were obtained from four children, 2-6 years of age, who had black stains on all erupted primary teeth. 16 different Actinomyces spp. were isolated, and antibiotic activity test with paper disc method was done. The results were as follows, 1. No.1 and No.5 Actinomyces spp. showed the antibiotic activity to Streptococcus mutans and the activity of No.5 Actinomyces spp. could compete with that of Oxacillin. 2. No.1 and No.5 Actinomyces spp. also exhibited the antibiotic activity to Bacillus cereus, Bacillus subtilis commonly used as experimental bacteria for testing antibiotic activity. 3. For identification of No.1 and No.5 spp., PCR analysis was done. No.5 spp. matched Actinomyces viscosus at 97% level but No.1 spp. didn't match.

  • PDF

Isolation and Identification of a Purple, Non-Sulfur Bacterium from Korea Coast

  • Cha, Mi-Seon;Kim, Gi-Han;Son, Hyeong-Sik;Lee, Na-Eun;Lee, Jeong-Eun;Jo, Sun-Ja;Lee, Sang-Jun
    • 한국생물공학회:학술대회논문집
    • /
    • 2003.04a
    • /
    • pp.409-411
    • /
    • 2003
  • A species of facultative photo-organotrophic, purple, non-sulfur bacterium was isolated from the west coast and the south coast 47 area of Korea at 2001 September. Separated 13 samples of changes with red color under $28{\sim}32\;^{\circ}C$, 3000 lux, anaerobe conditions for 7 days cultivated in Basal medium. For a pure isolation from 13 samples it used agar-shake tube method (0.4 % agar) and it separated 5 strains through 13-repetition test. The RAPD(Random Amplified Polymorphic DNA)-PCR result of strains (EGH-9, EGH-13, EGH-23, EGH-24, EGH-30) that EGH-24 and EGH-30 was same strain. For wastewater biodegradation test that 4 isolation strains cultivated in synthesis wastewater in 7 days. EGH-24 was high 63000 mg/L (CODcr) to 43400 mg/L (CODcr). EGH-24 was selected with efficient wast water treated strain. Based on the results obtained from morphology, nutrient requirements, major bacteriochlorophyll content, 165-rDNA phylogenetic analysis, this strain may be identified as a new strain of the genus Rhodobacter and named Rhodobacter sp. EGH-24.

  • PDF

Virulent Systemic Feline Calicivirus Infection in a Kitten (새끼고양이에서 발생한 고독성 칼리시바이러스(VS-FCV) 감염증)

  • Park, Jong-In;Suh, Sang-Il;Hyun, Changbaig
    • Journal of Veterinary Clinics
    • /
    • v.32 no.5
    • /
    • pp.445-448
    • /
    • 2015
  • A 5 month old Korean domestic short haired male kitten (weighing 1.7 kg) was presented with primary complaints of upper respiratory disease (URD) signs and skin ulceration and edema on face, feet and footpad with lameness. Diagnostic test revealed leukopenia, lymphopenia, pancreatitis and feline calicivirus (FCV) infection. Diagnosis of virulent systemic FCV (VS-FCV) was made on clinical signs, isolation of calicivirus via PCR and exclusion of other causes of acute upper respiratory disease. Therapeutic strategies were directed to lessen URD signs and to treat secondary bacterial infection and antiviral infection. One month after this therapy, skin lesions on face and feet and URD signs were much improved, although the lameness persisted mildly. To author's best knowledge, this is the first case reporting VS-FCV infection in a kitten in Korea.

Identification of Tricholoma matsutake in a Pine Root by ITS Region Amplification and RAPD Analysis with Different Mycorrhiza (ITS 영역 증폭에 의한 소나무 송이균 뿌리 감염 확인 및 RAPD에 의한 타 균근과의 비교)

  • Kim, Myungkil;Ryu, Sun-Hwa;Bak, Wonchull;Park, Hyun;Ka, Kang-Hyeon;Sohn, Hee-Kyung
    • Journal of the Korean Wood Science and Technology
    • /
    • v.34 no.6
    • /
    • pp.96-103
    • /
    • 2006
  • A simple method for identifying of ectomycorrhizal fungi was presented, using the polymerase chain reaction (PCR) to amplify the ITS (Internal transcribed spacer) regions of the nuclear ribosormal repeat. The sequences analyzed 6 species, Pisolithus tinctorius, Chroogomphus rutilus, Leucogyrophana pinastri, Suillus granulatus, Lactarius laeticolorus, and Suillus bovinus at hongreung forest, and analysed 10 species, Craterellus lutescens, Thelephoroid mycorrhizal, Lactarius quieticolor, Tricholoma matsutake, Lactarius chrysorrheus, Sarcodon aspratus, Russula versicolor, Suillus luteus, Tricholoma terreum, and Amanita vaginata at hongcheon forest. Finally, the amplification by PCR with ITS1-ITS4 primers offers good results over classical identification for ectomycorrhizal fungi species.

Molecular identification of Mariner Transposase-like Element from Four silkmoths (견사곤충에서 Mariner Transposase-like Element의 분자적 동정)

  • Lee, Jin Sung;Hwang, Jae Sam;Kim, Yong Sung;Suh, DongSang
    • Journal of Life Science
    • /
    • v.8 no.4
    • /
    • pp.457-464
    • /
    • 1998
  • As a first step for developing universal genetic transformation vector of dilkmoths., we identified the presence of mariner-like element(MLE) which is one of transposable element discovered from many insects to human species, from Bombyx mori, Bombyx mandarina, Antherae yammamai and Antherae pernyi. We used a degenerative primer pair designed from a transposase gene of Drosophila mauritiana and Hyalophora ceropia MLE. As results, major PCR product of 500bp expected as a part of transposase of MLE was detected from all the slkmoths used of this study using these primer. And hybridization assay using pBmoMAR as a probe DNA that was previously cloned from Bombyx mori by the same primer pair, confirmed the presence of MLE from all the silkmoths. This assay showed also that the endogenous MLE in genome of the silkworm is present as high copy number unlikely Drosophila mauritiana which has 10-20 copy number. This data will be a fundamental genetic information for developing mariner-derived vector to transform the silkmoths and other useful insects.

  • PDF

Selection of Molecular Biomarkers Relevant to Abnormal Behaviors of Medaka Fish (Oryzias latipes) Caused by Diazinon (다이아지논에 의해 야기된 송사리의 이상행동 연관 분자생물지표의 선발)

  • Koh, Sung-Cheol;Shin, Sung-Woo;Cho, Hyun-Duk;Chon, Tae-Soo;Kim, Jong-Sang;Lee, Sung-Kyu
    • Environmental Analysis Health and Toxicology
    • /
    • v.24 no.4
    • /
    • pp.321-332
    • /
    • 2009
  • 본 연구의 목적은 다이아지논(Diazinon; O, O-diethyl O-[6-methyl-2 (1-methylethyl)-4-pyrimidinyl] phosphorothioate)에 노출된 모델 생물체(송사리)의 행동변화와 관련된 분자생물학적 기전 규명을 통하여 비정상적 행동의 모니터링을 위한 생물지표(biomarker)를 개발하는데 있다. 이를 위해 우선 suppression subtractive hybridization (SSH) 및 DNA microarray 기법을 활용하여 다양한 유전자를 스크리닝하였다. 다이아지논에 노출시킨 송사리에서 발현의 차이가 나는 상향 조절된 유전자 97개 (알려지지 않은 유전자 27개 포함)와 하향 조절된 유전자 99개 (알려지지 않은 유전자 60개 포함)를 동정 하였고 이들 중 이상행동과 관련되는 것으로 보이는 유전자 10개 (상향조절 5개, 하향조절 5개)를 선발하였다. 이들 중에서 primer 제작이 잘된 beta-1, Orla C3-1, parvalbumin 및 apolipoprotein E을 선발하여 그 유전자 발현을 real-time PCR 기법을 사용하여 정량적으로 모니터링 하였다. Orla C3-1, parvalbumin 및 apolipoprotein E는 고농도의 다이아지논 처리(1000 ppb; 24 h)에서 그 발현이 억제됨이 관찰되었다. 다이아지논 처리 시 신경질환 (알츠하이머 병 및 다운신드롬)에 관련된 apolipoprotein E와 근육세포의 유연화에 작용하는 parvalbumin 등의 발현억제는 송사리의 인지능력 교란 및 근육세포의 경직 등을 각각 유도하여 송사리의 비정상적 행동을 야기하는 것으로 판단되었다. 따라서 이들 생물지표는 신경독성물질에 의한 송사리 및 기타 어류의 이상행동의 변화의 감지에 활용될 수 있을 것으로 사료된다.

Root Rot of Japanese Angelica Caused by Phytophthora cactorum in Nursery and Mycological Characteristics of the Isolates (두릅나무 묘목생산포의 역병 발생 및 분리균의 균학적 특성)

  • Lee, Sang-Hyun;Lee, Jae-Pil;Kim, Kyung-Hee;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
    • /
    • v.33 no.2
    • /
    • pp.98-102
    • /
    • 2005
  • In 2003 to 2005, the root rot of Japanese angelica (Aralia elata) was surveyed in nursery beds of Korea, where incidence of the disease often reached up to 100%. Three isolates were obtained from the infected roots, and identified as Phytophthora cactorum on the basis of cultural, morphological characteristics and molecular analysis. The isolates were characterized by having markedly papillate and broadly ovoid deciduous sporangia. The optimum temperature for mycelium growth was at $25^{\circ}C$ on V8 juice agar. Pathogenicity of the isolates was confirmed by soil mixture inoculation. Approximately 900 bp of ITS rDNA was amplified from all 3 isolates and band pattern of restriction fragments observed by Alu I, Msp I, and Taq I digestion also supported the result of the morphological identification when compared with PhytID database.

Characterization of a Ligninase Producing Strain, Serratia marcescens HY-5 isolated from Sympetrum dopressiusculum (고추좀잠자리 (Sympetrum depressiusculum)로부터 분리한 리그닌 분해균주, Serratia marcescens HY-5의 특성)

  • Kim, Ki-Duck;Park, Doo-Sang;Shin, Dong-Ha;Han, Bo-Na;Oh, Hyun-Woo;Youn, Young-Nam;Park, Ho-Yong
    • Korean journal of applied entomology
    • /
    • v.45 no.3 s.144
    • /
    • pp.301-307
    • /
    • 2006
  • A ligniolytic bacterial strain was isolated from the digestive tract of a red dragonfly, Sympetrum dopressiusculum. It was identified as a Serratia marcescens HY-5 by 16S rDNA sequence analysis and physiological and biochemical analysis. The isolated strain showed proportional increase of ligninolytic activity to the cell growth in the culture media which include lignin compounds. It showed about 25-45% decomposition of lignin compound by 48 hr incubation especially, showed effective decomposition of monomer lignin compounds, vanillin and guaiacol, and a dimer, dealkaline lignin. PCR amplification of 16S rDNA followed by denaturing gradient gel electrophoresis analysis showed high density of S. marcescens HY-5 in the gut of the S. depressiusculum at both gut samples which collected at different site.

Isolation of Coat Protein Gene from Cucumber Mosaic Virus and Its Introduction into Tobacco (오이 모자이크 바이러스 외피 단백질 유전자 분리 및 담배로의 형질전환)

  • 손성한;김경환;김영태;박종석;김주곤;이광웅;황영수
    • Korean Journal of Plant Tissue Culture
    • /
    • v.22 no.3
    • /
    • pp.149-155
    • /
    • 1995
  • The coat protein (CP) gene was cloned from RNA genome of the Cucumber Mosaic Virus strain ABI (CMV-ABI) isolated in Korea. The comparisons of the nucleotide sequence of the cloned CP gene and its deduced amino acid sequences with other CP genes revealed that the CMV-ABI belongs to subgroup I (type I), CMV-ABI developed the typical mosaic symptom in infected plants. Tobacco plants (Samsun and NC82) were transformed by leaf-disc transformation via Agrobacterium, temefaciens LB4404 harboring pVCP, witch CMV-ABI CP gene was inserted into the pBI121, and a number of mature transgenic tobacco plants were developed. Southern and PCR analysis of genomic DNA from the transgenic plants showed that the CP gene was integrated into the genomes of the most of the transgenic plant. Result of the segregation patterns of resistance in T1 seedlings of the plants to kanamycin showed that the transgenic plants containing l,2 and 3 copies of CP gene were50%, 39% and 11% of the total transgenic plants, respectively.

  • PDF