• Animal Systematics, Evolution and Diversity
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• v.11 no.4
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• pp.469-477
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• 1995

Drosophila lacertosa and D. sordidula are members of the robusta species group in virilis section of Drosophila. The mtDNA of both species was analyzed, using 10 restriction endonucleases. The mtDNA genome size of D. lacertosa and D. sordidula was 15.7 kbp, altogether, and the numbers of mtDNA fragment were 26 and 32, respectively. Restriction cleavage map of mtDNA in these species was constructed. The patterns of cleavage map were very different between two species and it means that speciation was taken for a long time ago.

• Korean Journal of Microbiology
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• v.23 no.1
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• pp.13-19
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• 1985

Acc I endonuclease has been isolated from 300g (wet weight) cells of Acinetobacter calcoaceticus. The cells were broken by using French press at 20, 000p.s.i. After ammonium sulfate fractionation, the enzyme was further purified by heparin agarose, DEAE-sephades, Affi.-gel Blue, phosphocellulose, and hydroxylapatite column chromatography. The purified Acc I endonudlease has a single polypeptide species and its subunit molecular weight was 45,000 ${\pm}$ 1,000 daltons as judged by 10% SDS-polyacrylamide gel electrophoresis. The isolated enzyme was essentially free of contaminating nucleases as judged by homochromatography by using a $^{32}P-labeled$ oligonucleotide. The enzyme showed maximum activity at pH values between 8.0 and 11.0 and in the presence of $MgCl_2$. Acc I endonuclease was maximally active in the absence of NaCl and was completely inhibited at 200 mM NaCl.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.138-146
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• 1985

A Bacillus licheniformis ATCC31667 gene coding for a glucose isomerase has been cloned and expressed in glucose isomerase negative mutant of Escherichia coli. A recombinant plasmid, constructed by ligation of a EcoRI fragment of B.licheniformis chromosomal DNA to vector plasmid pBR322, was expressed glucose isomerase positive in E.coli LE392-6 with growth on minimal medium containing xylose as a sole carbon source. This recombinant plasmid, designated pBGI6, had the insery of 4.1Kb of Bacillus gene in EcoRI site, and restriction map of the plasmid was established. The plasmid pBG16 was very stable after 10days of serial transfer to a fresh medium. The activity of glucose isomerase from the transformed cell containing pBGI6 was increased about 20 fold than its wild type of host.

• Korean Journal of Microbiology
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• v.24 no.1
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• pp.18-23
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• 1986

A new type II restriction endonuclease, Bdi I, has been isolated from Brenibacterium divaricatum FERM 5948 by procedures of ammonium sulfate fractionation, DEAE-cellulose chromatography and heparin agarose chromatography. The purified Bdi I restriction endonudlease had the same cleavage patterns of Cla I whose recognition sequence is 5' ATCGAT 3'. From the result that ${\lambda}-Cla$ I DNA frahment could be cloned in pBR 322 digested with Bdi I, it has been proven that Bdi I cuts between T and C(5' AT/CGAT3') within the recognition sequence and produces 5'pCG cohesive end. The optimal temperature for the Bdi I restriction endonuclease activity was $37^{\circ}C$, and optimal salt (NaCl) concentration was 50-100 mM.

• Korean Journal of Microbiology
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• v.32 no.4
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• pp.297-300
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• 1994

About thirty bacterial strains of actinomycete isolated from the soil were examined for the presence of restriction endonuclease activity. Streptomyces diastatochromogenes, which was identified previously, was found to contain restriction endonuclease activity. The purification of this enzyme, SdiI, was carried out via streptomycin sulfate precipitation and ammonium sulfate fractionation followed by hydroxylapatite column chromatography. Sephacryl S-200 HR column chromatography and second hydroxylapatite column chromatography. SDS-polyacrylamide gel electrophoresis of the active protein (purified from various column chromatography) resulted in 35,000 Da protein.

• Microbiology and Biotechnology Letters
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• v.38 no.4
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• pp.455-460
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• 2010

A polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis was applied to detect and identify ten Weissella spp. frequently found in kimchi. The previously reported genus-specific primers designed from 16S rDNA sequences of Weissella spp. were adopted but PCR was performed at the increased annealing temperature by $4^{\circ}C$. The sizes of amplified PCR products and restricted fragments produced by AluI, MseI, and BceAI endonucleases were well correspond with the expected sizes. W. kandleri, W. koreensis, W. confusa, W. minor, W. viridescens, W. cibaria, and W. soli were distinguished by AluI and MseI and W. hellenica and W. paramesenteroides were identified by BceAI. W. thailandensis was distinguished when restriction pattern of other species was compared but identified by the single use of MspI.

• Journal of Ginseng Research
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• v.27 no.3
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• pp.146-150
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• 2003

This study was carried out to obtain basic information on breeding using PCR-aided RFLP technology which can identify the variation inter- and intra-species of ginseng in the level of DNA. It was intended to investigate banding pattern on psbA and rbeL genes of chloroplast DNA in ginseng after treating with restriction enzymes. To isolate psbA and rbcL genes of chloroplast, both psbA-N, psbA-C primer and rbcL-N, PX-1 primer were used. As a result, 1,008 bp band of psbA gene and 1,336 bp band of rbcL gene were appeared, which was optimal and expected molecular weight. In addition, primers to isolate atpB, rpoB, trnL, and trnF genes were used, resulting in the expected 1366, 900, 1500 and 1008 bp bands. Genes of psbA and rbcL isolated by PCR were cut by restriction enzymes, Sau3A, TaqI, AluI, HaeIII, and RFLP pattern was investigated. KG line and other species of ginseng were cut by TaqI treatment, and bands were located in 800 bp. The treatment treated by AluI also showed the same 800 bp band in KG line and other species. In HaeIII treatment, 500 bp of faint bands were shown in case of KG line, whereas any bands were not observed in other species. All chloroplast genes formed bands by PCR amplification. However, it was not evident to distinguish intra-or inter-species of ginseng after restriction enzyme treatment. Therefore, more restriction enzyme treatment or sequence comparison method should be considered for further experiment.

• Korean journal of applied entomology
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• v.29 no.2
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• pp.132-135
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• 1990

Three mitochondrial cDNA probes from Aedes albopictus were used to demonstrate restriction site polymorphism in mtDNA of three sibling species of Anopheles quadrimaculatus(Say). It was shown by DNA hybridization to have substantial sequence homology betwen the mtDNA of different genus. The proves reveled local restriction site variation between members of the Anopheles quadrimaculatus sibling species complex. Mitochondrical DNA (mtDNA), isolated from individual mosquitoes was digested by type II restriction enzymes and four enzymes were found to be useful for the purpose. Hind III alone could be used to obtain a diagnostic restriction pattern.

• Journal of Life Science
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• v.22 no.2
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• pp.245-250
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• 2012

The 16S rDNA - RFLP types for six Vibrio species (V. fluvialis, V. proteolyticus, V. vulnificus, V. mimicus) including two core group members, V. alginolyticus and V. parahaemolyticu s, and Grimontia (Vibrio) hollisae were determined using PCR-RFLP analysis. Six tetrameric restriction enzymes (Alu I, Cfo I, Dde I, Hae III, Msp I, and Rsa I) were selected for RFLP analysis. V. alginolyticus, V. parahaemolyticus, and V. proteolyticus showed the same RFLP pattern following digestion with four of the six used restriction enzymes: CfoI, DdeI, MspI, and RsaI. Various restriction enzyme combinations generated digests recognizable as distinct RFLP types for each of the assayed Vibrio species. In particular, AluI single digestion produced species specific band patterns that enabled the differentiation between these Vibrio species. Dendrogram based on restriction patterns showed that two Vibrio core group members, V. alginolyticus and V. parahaemolyticus were closely related having a similarity over 90%. Although the observed RFLP pattern for Grimontia hollisae shared several common bands with other Vibrio spp., G. hollisae results were still clearly distinct from Vibrio spp. RFLP types for all restriction enzymes tested. If restriction enzymes are aptly selected, PCR-RFLP analysis is still a rapid and effective tool for differentiating Vibrio species.

• Korean Journal Plant Pathology
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• v.12 no.1
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• pp.91-98
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• 1996

벼도열병균 14개 균주와 벼 이외 화본과 식물 도열병균 12개 균주를 대상으로 rDNA의 ITS II 부위를 증폭하여 그들의 핵산 구조 차이를 분석함으로 도열병균 균주간 분류를 시도하였다. 5.8S rDNA의 3`-말단 부위와 28S rDNA의 5`-말단 부위의 sequence 중 5`-CCCGGGAATTCGCATCGATCGATCGAATGAAGA-ACGCAGC-3`와 5`-CCCGGGATCCTCCGCTTATT-GATATGC-3`를 이용하여 PCR 증폭을 하였을 때 벼도열병균 14개 균주는 동일한 길이의 단일 밴드를 형성하였으며 벼 이외 화본과 식물 도열병균에서는 레드톱 식물로부터 분리한 도열병균만이 나머지 균주보다 38bp가 더 큰 길이를 가진 밴드를 형성하였다. PCR로 증폭된 DNA를 HaeIII와 MspI 제한효소로 절단하였을 때 벼도열병균 레이스간에는 제한효소 절단에 의한 전기영동 밴드 형태 차이를 관찰할 수 없었으나, 벼 이외 화본과 식물 도열병균 12개 균주는 3군으로 구분할 수 있었다. 벼도열병균 90=054와 강아지풀에서 분리한 도열병균 G90-5, 기장에서 분리한 G88-4, 바랭이에서 분리한 G88-5 그리고 레드톱에서 분리한 RT 균주의 ITS II 부위의 DNA 염기서열 비교 분석에 의하면 G88-4와는 다른 HaeIII와 MspI 제한효소 위치를 가지고 있었기에 제한효소 절단에 의한 전기영동 형태가 상이하였다. 또한 RT균주는 HaeIII와 MspI위치가 존재하지 않았다.