• 한국미생물·생명공학회지
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• 제15권5호
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• pp.352-355
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• 1987

새로운 제한효소인 Cth I을 Clostridium thermocellum ATCC 27405로부터 분리하여, 그 효소의 생화학적 특성을 연구하였다. 이 효소는 Bcl I endonuclease의 isoschizomer로서 5'-TGATCA-3'를 인식한다. Cth I endonuclease는 섭씨 60도의 높은 온도에서 잘 작용하며, 10mM까지의 저농도의 NaCl과 MG$^{2+}$ 이온의 존재하에서 pH7.5와 10.5사이에서 최적의 활성을 보여주었다. Cth I endonuclease의 활성은 DNA 기질이 dam methylation 되었을 때 저해를 받았다.

• 미생물학회지
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• 제25권1호
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• pp.73-79
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• 1987

A restriction endonuclease, Kpn I has been isolated from Klebsiella pneumonia. Cells were broken by sonication. After ultracentrifugation the supernatant containing Kpn I activity was further purified by Sepharose-6B gel filtration, DEAD-Cellulose, Heparin-Agarose, and Aminohexyl-Agarose column chromatography. Final enzyme preparation was essentially free of contamination exonuclease and phosphatase, as judged by ligation-recut test. Total activity of the enzyme recovered from 10 grams of cells was $4.7\times 10^5$ units.

• 미생물학회지
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• 제24권4호
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• pp.406-410
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• 1986

The isolation and charateriation of a type II restriction endonuclease from Xanthomonas citri IFO 3835 were described. This enzyme (Xci I endonuclease) is an isoschizomer of Sal I endonuclease recognizing 5'-GTCGAC-3' and cleaving at the site indicated by the arrow. Unlike Sal I endonuclease, Xci I endonuclease requries a NaCl concentration of 50mM for its maximum activity.

• 미생물학회지
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• 제32권4호
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• pp.297-300
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• 1994

토양에서 분리한 방선균에서 제한효소 활성을 조사하여 그 가운데 Streptomyces diastatochromogenes로부터 제한효소 활성을 확인하였다. 이 균주의 제한효소 SdiI을 균체 15g으로부터 streptomycin sulfate침전, ammonium sulfate 침전, hydroxylapatite colume chromatography, Sephacryl S-200 HR column chromatography, hydroxylapatite column chromatography의 단계를 거쳐 분리하였고, SDS-polyacrylamide gel-electrophoresis에 의하면 소단위체 분자량은 약 35,000Da으로 추정되었다.

• 대한수의학회지
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• 제33권1호
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• pp.43-51
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• 1993

Mitochondrial DNA(mt DNA) of Mammalian is the circular one which the 16.5K base pairs and show the maternal inheritance. Evolutional speed of nucleotide sequence is very fast. So that polymorphic analysis of mt DNA provide the useful informations to investigate the genetic relations of interspecies. Authors trials were focussed to compare with the polymorphic differences of mitochondrial DNA between Jindo and Japanese mongrel dogs. DNA was extracted from bloods of 21 head of Jindo dogs and 20 head of Japanese dogs and isolated using 10 kinds of restriction endonucleases(Apa I, BamH I, Bgl II, EcoR I, EcoR V, Hinc II. Hind III, Pst I, Sty I, Xba I) and then separated by the agarose gel electrophoresis. After sourthern blotting hybridization was completed using the mtDNA of Japanese mongrel dogs as a probe. Autoradiography was used to compare the polymorphism of mtDNA both dogs. The results obtained were as follows; 1. mt DNA of Jindo dog showed polymorphism resulting cleavage with four kinds of restriction endonuclease, Apa I, EcoR V, Hinc II, Sty I. While in the Japanese mongrel dogs observed the polymorphism in the five kinds of restriction endonuclease supplemented with EcoR I. 2. Compared with both dogs the frequency differences of DNA polymorphism were recognized in the specific restriction endonuclease Apa I. Consequently in the restriction endonuclease Apa I both dogs classified with three types as A, B, C however in the Jindo dogs frequency of C type was 71.5 percent but in Japanese mongrel dogs observed 45 percent in the A type. 3. DNA polymorphism obtained from the use of five kinds of restriction endonuclease were classified with seven types. In Jindo dogs frequency was highest in the type 6 as 71.4 percent but in the Japanese mongrel dogs showed 35 percent in the type 5. 4. Genetic distances calculated by NEI method showed 0.0089 in Jindo dogs and was 0.0094 in the Japanese mongrel dogs.

• 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.

• 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.

• BMB Reports
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• 제29권2호
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• pp.99-104
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• 1996

Site-directed substitutions were made to change the Ile91 of restriction endonuclease EcoRV to either Val, Ala or Gly to identify the role of Ile91 in recognition and catalysis, since substitution of Ile91 with Leu afforded dramatic effects on the activity and properties of restriction endonuclease EcoRV. These changes alter the size of the hydrophobic side chain at position 91 and thus might have revealed the reason for the altered phenotype of Ile91Leu. However, the properties of Ile91Val and Ile91Ala mutants were much like wild type EcoRV, in both activity and metal ion preference. Ile91Gly had very little activity with either $Mg^{2+}$ or $Mn^{2+}$ as cofactors. To try to understand the unusual $Mn^{2+}$ profile of the Ile91Leu mutant, two double mutants, Ile91Leu;Asp90Asn and Ile91Leu;Glu45Met were created. Both double mutants were seriously disabled by the second amino acid change. Ile91Leu;Glu45Met had some residual activity in the $Mn^{2+}$ reaction buffer, whereas the Ile91Leu;Asp90Asn displayed no detectable activity.

• 한국미생물·생명공학회지
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• 제22권5호
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• pp.485-490
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• 1994

A restriction endonuclease, PsyI, has been isolated from Pseudomonas syringae pv. pha- seolicola, and its catalytic properties have been studied. This enzyme was purified through strepto- mycin sulfate and ammonium sulfate fractionation, phosphocellulose Pll, DEAE-cellulose, hydroxy- apatite and Sephadex G-100 column chromatography. It's molecular weight was about 50,000 dalton as determined by 7.5% polyacrylamide gel electrophoresis containing 0.1% SDS. In catalytic proper- ties, PsyI shows stable at wide ranges of pH between 7.0 and 10.0, of temperature between 30$\circ$C and 37$\circ$C, and its thermal stability is between 25$\circ$C, and 45$\circ$C, at the presence Of 10 mM MgCl$_{2}$-PsyI essentially require Na salt for enzyme reaction, is rather inhibited in the high Na salt concent- ration. The presence of 2-mercaptoethanol is absolutely required for the enzyme activity. This endonuclease, PsyI was determined to be an isoschizomer of SalI from the results of the restriction mapping and DNA sequencing.

• 한국미생물·생명공학회지
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• 제21권4호
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• pp.299-305
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• 1993

Numerical identification was carried out for an isolate of Streptomyces D2-5 producing a new restriction endonuclease Svi I. Fifty taxonomic unit characters were tested and the data were analyzed numerically using the TAXON program. The isolate was best matched to Streptomyces violochromogenes in the major cluster 18 of Streptomyces. Therefore, it was concluded that the isolate was identified to be a member of Streptomyces violochromogenes.