• Title/Summary/Keyword: restriction enzyme

검색결과 413건 처리시간 0.03초

Phycocyanin locus내의 DNA Polymorphism에 의한 한국산 Cyanobacteria의 유전적 다양성 (Genetic Diversity of Korean Cyanobacteria determined by DNA polymorphisms within the Phycocyanin Locus)

  • 박진숙;권주리;유순애
    • 미생물학회지
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    • 제36권4호
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    • pp.249-253
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    • 2000
  • Cyanobacteria의 광합성 보조색소인 phycocyanin의 PC operon(cpc gene)을 PCR로 증폭하고, 제한효소로 처리하여 RFLP pattern을 비교하였다. Intergenic spacer sequence를 포함한 cpc gene은 실험에 사용한 cyanobacteria 균주 모두에게 증폭되었으며, 산물의 size는 약 700 bp였다. PCR산물을 5종의 제한효소로 처리한 결과 AluI, MspI, HaeIII는 같은 속내으ㅐ 균주간에 동일한 pattern을 나타내어 속 구분이 가능하였으며 CfoI은 Anabeana와 Synechocystis속의 균주간에 구별되는 양상을 나타내어 속내 균주 구별에 유용하였다. Restriction enzyme profile에 의한 phenogram에서 Anabeana, Chlorogloea, Synechyhocystis는 각각 하나의 cluster를 형성하여 cyanobacteria의 분류에 PC-IGS의 RFLP pattern이 유용함을 알 수 있었다.

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제한효소 DNA 분석법에 의한 국내분리 렙토스피라균의 동정 (Restriction Endonuclease DNA Analysis of Leptospiral Field Isolates from Korea)

  • 장우현;김석용;서정선
    • 대한미생물학회지
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    • 제22권4호
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    • pp.463-471
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    • 1987
  • The genomes of leptospiral field isolates from Korea belonging to serogroup Icterohaemorrhagiae (21 strains) and serogroup Canicola (1 strain) were analysed and compared by restriction enzyme analysis with EcoRI and HindIII as digesting enzymes. One isolate belonging to serogroup Canicola showed the same pattern as serovar portlandvere. All 21 isolates belonging to serogroup Icterohaemorrhagiae showed almost same patterns as Leptospira serovar lai from China, But with very slight differences 21 isolates could be classified into 8 subtypes and these grouping seems to reflect the differences in epidemiological niche. And also the geographical data consisted with the grouping into 8 subtypes. According to our results, we concluded that the restriction endonuclease analysis of chromosomal DNA will be an accurate and reliable method to compare and classify pathogenic leptospires.

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제한효소인 Sdi I을 생성하는 Streptomyces 분리 균주의 수리동정 (Numerical Identification of a Streptomyces Strain Producing Restriction Endonuclease Sdi I)

  • 배무;서원나;송은숙;이계준
    • 한국미생물·생명공학회지
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    • 제22권2호
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    • pp.126-133
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    • 1994
  • Numerical identification was aplied for Streptomyces sp.264, an isolate producing a new restriction endonuclease Sdi I. The restriction enzyme would appear to be an isoschisomer of Xho I. Fifty taxonomic unit characters were tested and the data obtained were analyzed numerically by using the TAXON program. The isolate was identified to be the major cluster 19 of Streptomyces and best matched to S. diastatochromogenes. It was, therefore, concluded that the isolate was identified to be a member of Streptomyces diastatochromogenes.

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다제내성 황색포도상구균이 가지고 있는 테트라사이클린 내성 플라스미드의 동정 (Characterization of Tetracycline Resistance Plesmid of Multidrug-resistant Staphylococcus aureus)

  • 이대운;문경호
    • 약학회지
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    • 제39권1호
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    • pp.6-9
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    • 1995
  • The clinical isolate Staphylococcus aureus SA2 had four kinds of plasmids and was resistant to ampicillin, chloramphenicol, clindamycin, erythromycin, gentamicin, kanamycin, methicillin, streptomycin, tetracycline and tobramycin. Transformation experiment demonstrated that 4.44 kb plasmid(pKH6) encoded resistance to tetracycline. The cleavage map of pKH6 was determined by restriction enzyme mapping techniques. The cleavage map is given for EcoRV, HindIII, HpaI, HpaII, KpnI and Xbal. Restriction endonucleases BamHl, BglI, BGIII, BstEII, EcoRI, HaellI, PstI, PvuII, SalI, Smal, and Xhol have no site on this plasmid. The restriction map revealed extensive structural homology between pKH6 and pT181.

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PCR기법을 이용한 오이 모자이크 바이러스 개나리 분리주(CMV-Fk)의 동정과 구분 (Identification and Differentiation of Cucumber Mosaic Virus Isolated from Forsythia koreana (CMV-Fk) Using PCR Techniques)

  • 이상용;박선정;최장경
    • 한국식물병리학회지
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    • 제14권4호
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    • pp.308-313
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    • 1998
  • Reverse transcription and polymerase chain reaction (RT-PCR) techiniques were used to identification and differentiation of cucumber mosaic virus isolated from Forsythia koreana (CMV-Fk). RT-PCT used by two set of 20-mer primers one was CMV-common primers and another was CMV subgroup I-specific primers designed in a conserved region of the 3' end of CMV RNA3, amplified about 490 bp and 200 bp DNA fragments from CMV-Fk, respectively. CMV could be detected by RT-PCR at a dilution as low as 10-4 in forsythia crude sap extracts. Restriction enzyme analysis of RT-PCR products using EcoRI and MspI showed that CMV-Fk belonged to CMV subgroup I. But, analysis of RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR) showed heterogeneity of RNA3 between CMV-Fk and CMV-Y as a member of subgroup I.

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Subcloning of Nodulin 26 Wild Type(S262) and Phosphorylation Site Mutant(S262D) into the Yeast Expression Vector pYES2

  • Cha, Youn-Soo
    • Preventive Nutrition and Food Science
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    • 제2권1호
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    • pp.61-65
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    • 1997
  • Wild type nodulin 26(nod 26) cDNA(S262) and phodphorylation aite mutant(S262D) were constructed by a yeast expression system using pYES2 plasmids(pTES2-D262 and pTES2-S262D) were sc-reened by restriction mapping with BamHI of KpnI. S262 nod 26 contained a sreine residue at position 262 and S262D nod 26 contained the substitution mutation of serine to aspartic acid residue at position 262 were verified by automated floursent DNA sequencing.

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CYTOTOXICITY OF PATULIN AND ITS EFFECT ON THE LAMBDA DNA CLEAVAGE BY RESTRICTION ENDONUCLEASE

  • Lee, Kil-Soo
    • Toxicological Research
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    • 제7권2호
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    • pp.157-163
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    • 1991
  • The effect of patulin, a mycotoxin, on the growth of Escherichia coli cell was investigated. E. coli cell elongation usually shown in SOS-response for DNA repair was induced by 20 mg of patulin per ml. After staining the E. coli chromosome with fluorescence dye(DAPI, 4', 6-diamino-2-phenyl-indole), chromosomal DNA partitioning was not affected by patulin. The observation indicateds that patulin acts as a DNA damaging agent which is effective for E. coli cell elongation introduced by the inhibition of septum formation.

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홀스타인종과 한우에 있어서 BoLA-DRB3 유전자의 단일염기다형과 반수체 분석 (Analyses of single nucleotide polymorphisms and haplotypes of BoLA-DRB3 gene in Holstein and Hanwoo)

  • 정행진;유성란;라세둘;이준헌;도창희;류승희;상병찬
    • 농업과학연구
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    • 제38권1호
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    • pp.51-63
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    • 2011
  • BoLA (bovine leukocyte antigens) have been known as gene complex related with bovine diseases and immunological traits. This study was conducted to find out the characteristics of BoLA-DRB3 gene related to mastitis and BL(bovine leukocyte) from 280 cattle [193 animals of Holstein cattle and 87 animals of Hanwoo]. As a result, five PCR-RFLP types (b, d, e, f and g) using HaeIII restriction enzyme, three BstYI restriction patterns (b, d and e) and eight RsaI restriction types(b, d, f, I, j, n, o and w) were identified. Moreover, we identified new d' type ($197{\rightarrow}175$/22), having one more cutting site by BstYI enzyme than d type allele and n' type ($180{\rightarrow}169$/11) having one more cutting site by RsaI enzyme than n allele was additionally identified. Next, we identified 53 SNPs in BoLA-DRB3 exon2 from 280 cattle. SNP frequency and heterozygosity of Holstein and Hanwoo were investigated in all the SNP genotype. These results might be based on research for identifying marker associated with bovine diseases.

동물에서 분리된 Salmonella균의 병원성 관련 Plasmid에 관한 연구 (Virulence-associated plasmids of Salmonella spp. isolated from animals in Korea)

  • 최원필;정석찬
    • 대한수의학회지
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    • 제32권3호
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    • pp.369-376
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    • 1992
  • This paper dealt with plasmid DNA profile in 98 Salmonella(S) isolated from pigs and cattle sources in Taegu, Gyeongbook and Gyeongnam during the period from 1984 to 1987. Also we were studied for restriction enzyme analysis of the plasmid DNA, and mouse infection, Sereny test and normal setum resistance test in guinea pig for S typhimurium and S enteritidis harbored or cured 60 megadalton(Md) plasmid and 36 Md plasmid, respectively. Of the 13 Salmonella isolated from cattle, 7 Salmonella harbored one or more plasmids and molecular sizes of the large plasmids were 60 Md for S typhimurium and 36 Md for S enteritidis. Of the 85 Salmonella isolated from pigs, 47 Salmonella were confirmed as being one or more plasmids, and all the S typimurium stains harbored 60 Md plasmid. In enzyme digestion with 8 types of restriction endonuclease for 60 Md plasmid DNA of S typhimurium, cleavage patterns were varied to enzymes, and the DNA was segmented into 4 to 15 fragments. In restriction enzyme analysis of 36 Md plasmid DNA obtained from four strains of S. enteritidis, the DNA showed the same cleavage patterns obtained with Eco RI, Hind III and Bam H I, and was segmented into 3 to 5 fragments. In virulence for mice by measuring the 50% lethal dose ($LD_{50}$), the $LD_{50}$ values obtained for 60 Md virulence-associated plasmid harbored strains of S typhimurium and 36 Md virulence-associated plasmid of S enteritidis were up to $10^4$-fold lower than the values obtained for the plasmid-cured strains of the same serotype. Only the plasmid harbored strains were resistant to the bactericidal activity of 90% guinea pig serum, and only they gave positive responses in sereny test. We suggested that their plasmid DNA might be associated with virulence for mice.

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