• 대한미생물학회지
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• 제35권1호
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• pp.49-60
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• 2000

ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp..

• BMB Reports
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• 제36권4호
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• pp.349-353
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• 2003

To explore the application of DNA chip technology for the detection and typing of Human Papillomavirus (HPV), the HPV6, 11, 16 and 18 gene fragments were isolated and printed onto aminosilane-coated glass slides by a PixSys 5500 microarrayer as probes to prepare the HPV gene chips. HPV samples, after being labeled with fluorescent dye by restriction display PCR (RD-PCR) technology, were hybridized with the microarray, which was followed by scanning and analysis. The experimental condition for preparing the HPV gene chips was investigated, and the possibility of HPV genotyping using gene chips was discussed. The technique that was established in this study for preparing HPV gene chips is practical. The results of the present study demonstrated the versatility and inspiring prospect of using this technology to detect and genotype HPV.

• Journal of Plant Biology
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• 제39권1호
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• pp.49-55
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• 1996

본 연구는 마늘(Allium sativum L.)의 기초적인 유전적 특성을 파악하기 위해, 단양마늘을 대상으로 염색체 DNA의 반복서열의 양상을 확인하고, 고반복서열이 매우 빠르게 reassociation되는 특성을 이용하여 이들에 해당되는 부분을 분리하고, 클로닝하였다. 이들 고반복서열 클론의 게놈 내의 copy수는 대체적으로 $10^{5}~10^{7}$이었다. 이 중 일부 클론의 염기서열과 분석한 결과, G/C 함량은 25~40% 정도로 낮았고, 일부서열의 내부에서는 소단위의 염기서열이 반복배열되어 있었다. 단양을 비롯한 문경, 서산, 의성 품종 사이에서 해당 반복서열의 변이정도를 조사하기 위하여, 다섯종류의 고반복서열을 탐침으로 이들 품종 마늘에 대한 RELP(restriction fragment length polymorphism)분석을 한 결과 이들 서열의 유전적변이는 거의 나타나지 않았다.

• 한국치위생학회지
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• 제13권5호
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• pp.889-900
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• 2013

Objectives : Molecular biology techniques were employed to assess diversity of bacterial in children's dental caries. Methods : DNA of germs was extracted and the diversity of the 16S rRNA clones was analyzed by amplified rDNA restriction analysis and sequencing. The experimental samples were pit and fissure caries (PC), deep dentinal caries (DC), smooth surface caries (SC), and supragingival plaque (PQ) from 50 children of age less than 12 years old. The control group was healthy teeth supragingival plaque (HT). Thirty clones from each 16S rRNA clone library of 5 samples were randomly selected, thus a total of 150 clones were analyzed. Results : Amplified rDNA restriction analysis uncovered 18, 20, 11, 17, and 22 phylotypes from healthy teeth, pit and fissure caries, deep dentinal caries, smooth surface caries, and supragingival plaque, respectively. Sequencing analysis found the dominance of Actinomycs naeslundii and Fusobacterium nucleatum in the healthy teeth; Leptotrichia sp. in the pit and fissure caries; Actinomyces sp., Streptococcus mutans, and Rahnella aquatilis in the deep dentinal caries; Streptococcus mutans and Actinomyces sp. in the smooth surface caries; Enterobacter hormaechei and Streptococcus sanguinis in the supragingival plaque. Conclusions : Clonal analysis identified 6 phyla, 20 genera, and 51 species.

• 미생물학회지
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• 제55권2호
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• pp.103-111
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• 2019

DNA 메틸화는 유전체의 무결성의 유지 및 유전자 발현 조절과 같은 박테리아의 다양한 과정에 관여한다. Alphaproteobacteria 종에서 보존된 DNA 메틸 전이 효소인 CcrM은 S-아데노실 메티오닌을 공동 기질로 사용하여 $N^6$-아데닌 또는 $N^4$-시토신의 메틸 전이 효소 활성을 갖는다. Celeribacter marinus IMCC 12053는 해양 환경에서 분리된 알파프로테오박테리아로서 GpC 시토신의 외향고리 아민의 메틸기를 대체하여 $N^4$-메틸 시토신을 생산한다. 단일 분자 실시간 서열 분석법(SMRT)을 사용하여, C. marinus IMCC12053의 메틸화 패턴을 Gibbs Motif Sampler 프로그램을 사용하여 확인하였다. 5'-GANTC-3'의 $N^6$-메틸 아데노신과 5'-GpC-3'의 $N^4$-메틸 시토신을 확인하였다. 발현된 DNA 메틸전이 효소는 계통 발생 분석법을 사용하여 선택하여 pQE30 벡터에 클로닝 후 $dam^-/dcm^-$ 대장균을 사용하여 클로닝된 DNA 메틸라아제의 메틸화 활성을 확인하였다. 메틸화 효소를 코딩하는 게놈 DNA 및 플라스미드를 추출하고 메틸화에 민감한 제한 효소로 절단하여 메틸화 활성을 확인하였다. 염색체와 메틸라아제를 코드하는 플라스미드를 메틸화시켰을 때에 제한 효소 사이트가 보호되는 것으로 관찰되었다. 본 연구에서는 분자 생물학 및 후성유전학을 위한 새로운 유형의 GpC 메틸화 효소의 잠재적 활용을 위한 외향고리 DNA 메틸라제의 특성을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권12호
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• pp.1691-1695
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• 2005

The bovine lymphocyte antigen (BoLA)-DRB3 gene encodes cell surface glycoproteins that initiate immune responses by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the most polymorphic bovine MHC class II gene which encodes the peptide-binding groove. Since different alleles favour the binding of different peptides, DRB3 has been extensively evaluated as a candidate marker for associations with various bovine diseases and immunological traits. For that reason, the genetic diversity of the bovine class II DRB3 locus was investigated by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). This study describes genetic variability in the BoLA-DRB3 in Iranian Golpayegani Cattle. Iranian Golpayegani Cows (n = 50) were genotyped for bovine lymphocyte antigen (BoLA)-DRB3.2 allele by polymerase chain reaction and restriction fragment length polymorphism method. Bovine DNA was isolated from aliquots of whole blood. A two-step polymerase chain reaction followed by digestion with restriction endonucleases RsaI, HaeIII and BstYI was conducted on the DNA from Iranian Golpayegani Cattle. In the Iranian Golpayegani herd studied, we identified 19 alleles.DRB3.2${\times}$16 had the highest allelic frequency (14%), followed by DRB3.2${\times}$7 (11%). Six alleles (DRB3.2${\times}$25, ${\times}$24, ${\times}$22, ${\times}$20, ${\times}$15, ${\times}$3) had frequencies = 2%. Although additional studies are required to confirm the present findings, our results indicate that exon 2 of the BoLA-DRB3 gene is highly polymorphic in Iranian Golpayegani Cattle.

• Genomics & Informatics
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• 제1권1호
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• pp.50-54
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• 2003

Genome of an extreme thermophile, Thermus caldophilus GK24 has been analyzed to construct the genomic map. The genomic DNAs encapsulated in agarose gel were digested with SspI, EcoRI, SpeI, and HpaI restriction endonucleases, and then the resulting genomic DNA fragments were analyzed by pulsed-field gel electrophoresis. Its restriction map has been constructed by analyzing sizes of the restriction fragments obtained from both complete and partial digestions. The circular form of its genome was composed of about 1.98 Mbp and a megaplasmid. The genomic loci for the genes of xylose isomerase, thioredoxin, tRNA-16S rRNA, 23S rRNA, L5 ribosomal protein, ADP-glucose pyrophosphorylase, DNA-ligase, and Tca DNA polymerase were determined by both Southern hybridization and PCR.

• 미생물학회지
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• 제25권4호
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• pp.282-289
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• 1987

Pseudomonas putida KU816에서 분리한 플라스미드 pKU10의 여러가지 특성을 조사하고 그 제한 효소 지도를 작성하였다. pKU 10은 암펴설런, 테트라사이클린, 클로람페니콜에 대한 내성 유전자를 갖는 작은 R factor로서 마이토마이신 C에 의하여 큐어링 된다. 플라스미드의 크기는 9.4Kb로 측정되었다. pKU 10은 Pseudomonas와 E.coli블 숙주로 하였을 때 안정하게 형질 발현이 되다. 또한 pKU 10의 불화합성균은 IncP-I으로 조사되었다. Eco RI, Xho I. SaiI, BglII, SmaI은 pKU 10 DNA를 한 부위에서 자르고, Pst I은 두 부위, Hind Ill는 여섯 부위에서 자른다. 제한 효소 지도는 제한 효소를 이중, 삼중으로 완전 소화시키거나, 부분 소화시켜서 얻었다. pKU 10은 Pseudomonas속에서 유용한 클로닝 벡터호 이용된 것으로 기대된다.

• Fisheries and Aquatic Sciences
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• 제9권3호
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• pp.101-106
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• 2006

The cytochrome c oxidase subunit I (COI) gene region of mitochondrial DNA (mtDNA) was examined in four abalone species to estimate its utility as a genetic marker using restriction fragment length polymorphism (RFLP) analysis. The utility was evaluated in terms of genetic divergence and relationships among Haliotis discus hannai, H. rufescens, H. rubra, and H. midae in both hemispheres of the world. There was clear genetic divergence in the mtDNA COI region between all pairs of the four species. Moreover, relationships among the abalone species were reflected in their geographical distributions and morphological characteristics. Therefore, RFLP analysis of the mtDNA COI region is a suitable genetic marker for the estimation of genetic divergence and relationships among abalone species. However, it is not effective for the evaluation of genetic differences within abalone species.

• The Plant Pathology Journal
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• 제20권4호
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• pp.264-270
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• 2004

Fusarium oxysporum f. sp. fragariae is a fungal pathogen causing strawberry wilt disease. The random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphisms (RFLPs) of intergenic spacer (IGS) region of rDNA were used to identify genetic variation among 22 F. oxysporum f. sp. fragariae isolates. All isolates could be distinguished from each other by RAPD analysis and RFLP of 2.6 kb amplified with primer CNS1 and CNL12 for IGS region of rDNA. Cluster analysis using UPGMA showed eight distinct clusters based on the banding patterns obtained from RAPD and rDNA RFLP. These results indicate that F. oxysporum f. sp. fragariae isolates are genetically distinct from each other, There was a high level genetic variation among F. oxysporum f. sp. fragariae.