• 제목/요약/키워드: endonuclease

검색결과 247건 처리시간 0.017초

MLS계 항생물질 유도 내성 유전자의 크로닝과 유전자의 조절기전 -Streptococcus sp. TR-1에서 분리한 pMB 4 Plasmid의 MLS계 항생물질 유도내성- (Cloning of the MLS Antibiotics Inducible Resistance Gene and Its Control Mechanism -Inducible Resistance to MLS Antibiotics of pMB4 Plasmid Isolated from Streptococcus sp. TR-1-)

  • 정순학;곽진환;김희선;심미자;최응칠;김병각
    • 약학회지
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    • 제34권2호
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    • pp.139-146
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    • 1990
  • Streptococcus sp. TR-1 which has inducible resistance to MLS antibiotics was isolated from soil samples in Korea. Streptococcus sp. TR-1 was cultured in Lysis broth, then a plasmid was isolated by modified Elliker method. Bacillus subtilis UOTO277 was transformed with that plasmid. This result showed that the plasmid has the gene relating with inducible resistance to MLS antibiotics. It was named pMB4 and its size was determined about 2.4 Kb by results of digestion with various restriction enzymes. Restriction endonuclease cleavage site map of pMB4 plasmid was made by double digestion of the plasmid. pMB4 plasmid has different restriction endonuclease site map from the other plasmids that have been discovered in Streptococcus sp. so far. And it could be identified that pMB4 plasmid does not have homology with ermK of Bacillus licheniformis EMR but has homology with ermC of Staphylococcus aureus from the results of Southern hybridization.

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담배나방 핵다각체병바이러스의 형태 및 생화학적 특성 (Morphology and Biochemcial Characteristics of a Nuclear Polyhedrosis Virus Isolated from the Oriental Tobacco Budworm, Helicoverpa assulta (Guenee))

  • 진병래;박현우;우수동;김우진;김우진;박범석;강석권
    • 한국응용곤충학회지
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    • 제34권3호
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    • pp.218-223
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    • 1995
  • 담배와 고추의 주요해충인 담배나방 [Helicoverpa assulta(Guenee)]의 미생물적 방제법을 개발하기 위하여, 아직 국내에서 보고되지 않은 담배나방 핵다각체병 바이러스 담배나방 사충으로 부터 분리하고, 바이러스의 전자현미경 관찰 및 바이러스 DNA의 제한효소 분석 등을 통한 생화학적 특성을 조사하였다. 담배나방 핵다각체병 바이러스의 다각체는 구형에 가까운 20면체로서 크기는 평균 약 1.0$\mu$m 정도이고, 다수의 바이러스 입자가 다각체 단백질에 포매되어 있었다. 포매된 바이러스 입자는 한개의 봉상 nucleocapsid가 하나의 envelope 내에 매립된 형태의 SNPV(single embedded nuclear polyhedrosis virus)로, 형태는 전형적인 봉상으로서 그 크기는 약 $65nm\times300nm$ 였다. 또한 다각체 단백질의 SDS-PAGE 분석 결고, 분자량은 약 31 Kd이었으며, 담배나방 핵다각체병바이러스 DNA를 분리하여 EcoRI. HindIII 등 수종의 제한효소로 처리분석한 결과, 그 genome의 크기는 약 120Kb 정도였다. 본 핵다각체병바이러스는 담배나방 유충에만 병원성을 나타내 기주특이성을 보였다.

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국내 가금 유래 병원성 대장균의 분자역학적 분석 (Molecular epidemiologic analysis of pathogenic Escherichia coli isolated from poultry in Korea)

  • 성명숙;김진현;김기석
    • 대한수의학회지
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    • 제50권3호
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    • pp.239-246
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    • 2010
  • Among 203 avian pathogenic Escherichia coli (APEC) isolated from poultry with colibacillosis in korea, 14 isolates were selected from total 68 isolates transferred R plasmid and classified into 5 groups on the basis of antimicrobial minimal inhibitory concentration (MIC) pattern, farm source and O serotype. An association between clonal origin and R plasmid of them was investigated by R plasmid profile, restriction endonuclease analysis and pulsed-field gel electrophoresis (PFGE). The strains that showed the same or very similar antimicrobial MIC pattern, but different farm source and O serotype, revealed different PFGE pattern, which seemed to be different clonal origin. And the strains that showed the same MIC pattern and O serotype, revealed different PFGE pattern, seemed to be originated from different clone. Also the strains showing the same MIC pattern and farm source, but different O serotype, revealed to be different clonal origin. The strains that showed the same or similar MIC pattern, farm source, and O serotype, revealed identical or similar PFGE pattern, which seemed to belong to be one clone. Meanwhile, horizontal transfer of R plasmid seems to be common in APEC with regardless of O serotype and clone of the strains. These results indicate that rapid and accurate epidemiological survey of APEC can be possible by the combination of O serotyping, plasmid profiling and PFGE analysis following the classification of them into groups of antimicrobial drug resistance pattern.

수질환경에서 일어나는 항생물질 내성유전자의 전이와 재조합 (Transfer and genetic recombination of antibiotic resistance genes occurring in water environment)

  • 김치경;이성기;김영창
    • 한국미생물·생명공학회지
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    • 제14권3호
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    • pp.245-250
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    • 1986
  • 청주시 무심천의 하천수에서 항생물질에 내성을 나타내는 Gram 음성 세균을 분리하여 수질환경에서 일어나는 R 플라스미드의 전이를 연구하였다. 분리된 균주사이에서 접합에 의한 R 플라스미드의 전이는 실험실 환경에서 1.1$\times$$10^{-6}$-1.2$\times$$10^{-7}$, 하천의 수질환경에서 1.2$\times$$10^{-7}$-1.0$\times$$10^{-9}$으로 나타나, 자연의 수질환경에서도 R 플라스미드의 전이가 일어남을 확인하였다. 또 T-44 균주의 Ap$^{r}$Cm$^{r}$Tc$^{r}$ 플라스미드는 형질전환에 의하여 E. coli HB 101에 1.7$\times$$10^{-6}$의 비율로 전이되었다. 분자의 크기가 약 9.01kb로 측정된 Ap$^{r}$Cm$^{r}$Tc$^{r}$플라스미드 DNA를 제한효소로 처리한 결과 이 플라스비드에는 EcoRI과 BamHI의 절단부위가 각각 하나씩 존재하고 P-stI의 절단부위는 3개가 있었다.

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누에와 흰불나방 핵다각체병바이러스의 생화학적 특성 (Biochemical Characteristics of the Nuclear Polyhedrosis Viruses of the Fall Webworm, Hyphantria cunea, and the Silkworm, Bombyx mori)

  • 김현욱;박범석;진병래;임대준;강석권
    • 한국응용곤충학회지
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    • 제28권3호
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    • pp.105-112
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    • 1989
  • 누에(Bombyx mori)와 흰불나방(Hyphantria cunea)으로 부터 분리된 핵다각체병바이러스(nuclear polyhedrosis virus: NPV)를 동정하기 위하여 전자현미경 관찰한 결과, BmNPV 다각체의 크기는 $3 \mu\textrm{m}$ 정도의 18면체로 외형이 균일하였으나, HcNPV는 1.5-$2 \mu\textrm{m}$ 정도이며 부정형이었다. Alkaline protease를 부활화시킨 후 SDS-PAGE한 다각체 단백질의 分子물은 BmNPV가 30 KD, HcNPV는 31 KD인 major band와 이들의 중합체(polymer)로 생각되는 57 KD, 112 KD의 minor band들이 관찰되었다. 또한 virion 단백질을 SDS-PAGE한 후 은염색한 결과, BmNPV는 분자량 9.6~112 KD인 47개의 band, HcNPV 경우는 분자량 9.4~l11 KD인 48개는 band가 관찰되었다. BmNPV와 HcNPV DNA의 제한효소 처이에 의한 전기영동 패턴을 관찰했으며, 각각의 genome 크기는 BmNPV가 약 116.4 Kb, HcNPV의 약 114.6 Kb였다.

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Streptomyces diastatochromogenes로부터 분리된 SdiI의 특성에 관한 연구 (Characterization of a Restriction Endonuclease, SdiI from Streptomyces diastatochromogenes)

  • 배무;송은숙;황혜연;임정빈
    • 미생물학회지
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    • 제32권4호
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    • pp.301-305
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    • 1994
  • 토양분리 방선균 Streptomyces diastatochromogenes로부터 분리된 제한효소 SdiI은 넓은 범위의 pH(7.0~12.5)와 온도 ($-60^{\circ}C$)에서 활성을 보였으며, 고농도 (-500mM) NaCl이 있어도 작용하였다. 또한, $20~60^{\circ}C$ 온도에서 안정하며, 활성을 갖기 위해서는 $MgCl_2$를 필요로 하였다. Lambda DNA에 대한 지도 작성으로부터 XhoI의 isoschizomer로 추정되었으며, DNa 염기서열 분석 결과, 인식, 절단 서열은 다음과 같이 결정되었다. 5‘-C${\downarrow}$TCGA G-3' 3'-G AGCT${\uparrow}$C-5'

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Photoprotection by Topical DNA Repair Enzymes

  • Yarosh, Daniel B.
    • Journal of Photoscience
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    • 제9권2호
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    • pp.186-189
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    • 2002
  • Many of the adverse effects of solar UV exposure appear to be directly attributable to damage to epidermal DNA. In particular, cyclobutane pyrimidine dimers (CPD) may initiate mutagenic changes as well as induce signal transduction responses that lead to a loss of skin immune surveillance and micro-destruction of skin structure. Our approach is to reverse the DNA damage using prokaryotic DNA repair enzymes delivered into skin using specially engineered liposomes. T4 endonuclease V encapsulated in liposomes (T4N5 liposome lotion) enhanced DNA repair by shifting repair of CPD from the nucleotide excision to the base excision repair pathway. Following topical application to humans, increased repair limited UV-induction of cytokines, many of which are immunosuppressive. In a recent clinical study, topical treatment of UV-irradiated human skin with T4N5 liposome lotion reduced the suppression of the nickel sulfate contact hypersensitivity response. Similarly, the photoreactivating enzyme enhances repair by directly reversing CPDs after absorbing activating light. Here also treatment of UV-irradiated human skin with photoreactivating enzyme in liposomes and photoreactivating light restored the response to the contact allergen nickel sulfate. These findings confirm in humans the observation in mice that UV induced suppression of contact hypersensitivity is caused in part by CPDs. We have tested the ability of T4N5 liposome lotion to prevent UV-induced skin cancer in patients with xeroderma pigmentosum (XP), who have an elevated incidence of skin cancer resulting from a genetic defect in DNA repair. Daily use of the lotion for one year in a group of 20 XP patients reduced the average number of actinic keratoses by 68% and basal cell cancers by 30% compared to 9 patients in the placebo control group. Delivery of DNA repair enzymes to skin is a promising new approach to photoprotection.

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Cloning of Steroid $\Delta^1$-dehydrogenase Gene of Arthrobacter simplex IAM 1660

  • Bae, Moo;Bae, Song-Mee;Lee, Mi-Kyung;Lee, Jeong-Kug
    • Journal of Microbiology and Biotechnology
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    • 제6권2호
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    • pp.142-144
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    • 1996
  • To clone the gene coding for steroid $\Delta^1$-dehydrogenase of Arthrobacter simplex, its genomic library was constructed with a , $\lambda$gt11 expression vector and immunoscreened with antiserum against the enzyme. One positive clone was found to carry a 1.6-kb EcoR I restriction endonuclease fragment of A. simplex DNA. The restriction map of the 1.6-kb EcoR I fragment was determined after cloning of the DNA into pBS vector.

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소 허피스바이러스 gIII 유전자 크론닝 및 발현 (Cloning and Expression of Bovine Herpesvirus-1 gIII of Korean Isolate PQ Strain)

  • 권창희;민부기
    • 대한바이러스학회지
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    • 제26권2호
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    • pp.173-179
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    • 1996
  • The gene encoding gIII of bovine herpesvirus type 1 (BHV-1) PQ strain was cloned and expressed in baculovirus. Although the gIII gene is located in Hind III I fragment as the case of the other BHV-1 strains, differences in size and restriction endonuclease site within the fragment were identified. The gIII expression was predominantly detected on the surface on insect cells by indirect immunofluoresecnce assay using monoclonal antibody. The western blotting analysis also revealed the presence of expressed protein of a similar molecular size to the original gIII protein. The immunogenicity of expressed protein were tested in guinea pigs. The immunized guinea pigs with expressed protein developed the neutralizing antibodies against BHV-1.

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대기부유분진추출물로 야기된 DNA 손상에 대한 Quercetin, Vitamin C 및 Trolox 의 보호효과 (Chemopreventive Effect of Quercetin, Vitamin C and Trolox Against the Organic Extract of Airborne Particulate Matter Induced Genotoxicity in A549 Human Lung Carcinoma Cells)

  • 김남이;허문영
    • 약학회지
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    • 제51권4호
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    • pp.239-245
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    • 2007
  • In order to evaluate the genotoxicity of airborne particulate matter extracted with dichloromethane (APE), the rat microsome mediated (S-9) or DNA repair enzyme treated Comet assays were performed using the single cell gel electrophoresis in A549 human lung carcinoma cells. It was found that the cells interacting with APE showed more DNA single-strand breaks relative to untreated cells. The genotoxicity of APE was increased with the treatment of S-9 mixture. Microsome mediated DNA damage was inhibited by CYP1Al inhibitor, quercetin. The APE also showed oxidative DNA damage evaluated by endonuclease III treatment. Oxidative DNA damage of APE was inhibited by antioxidants such as vita- min C and Trolox. We also found that the vegetables or fruits extract may reduce APE-induced genotoxicity by their anti- oxidant activity and CYP1A1 inhibition.