• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.120.2-120
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• 2003

Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. In Korea, grapevine variety (GeoBong) have severely been infected by the bacteria since stems of the variety were buried in soil for overwintering. Infection ratio over 70-80％ was observed on 7 years old GeoBong grapevine in Ansung and Cheonan. PCR specific primers for A. vitis strains were designed using nucleotide sequences of vir A gene in Ti-Plasmid, pheA gene in chromosomal DNA and a URP-PCR polymorphic band. Three hundred bacterial strains were isolated from the different 80 galls formed on GeoBong grapevine in Cheonan and Ansung of Korea and were screened to identify A. vitis using the three specific PCR primers for Agrobacterium vitis. Twenty-four bacterial strains that are detected by the primers were further confirmed by pathogenicity and biochemical methods. To investigate the genomic diversity of the bacterial strains, twenty primers of 20 mer referred to universal rice primers (URP) were applied for PCR fingerprinting, Of them, URP2R and URP2F primers could effectively be used to detect polymorphism within the bacterial strains.

• Tuberculosis and Respiratory Diseases
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• 제77권4호
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• pp.172-177
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• 2014

Background: Pseudomonas aeruginosa infection is particularly associated with progressive and ultimately chronic recurrent respiratory infections in chronic obstructive pulmonary disease, bronchiectasis, chronic destroyed lung disease, and cystic fibrosis. Its treatment is also very complex because of drug resistance and recurrence. Methods: Forty eight cultures from 18 patients with recurrent P. aeruginosa pneumonia from 1998 to 2002 were included in this study. Two or more pairs of sputum cultures were performed during 2 or more different periods of recurrences. The comparison of strains was made according to the phenotypic patterns of antibiotic resistance and chromosomal fingerprinting by pulsed field gel electrophoresis (PFGE) using the genomic DNA of P. aeruginosa from the sputum culture. Results: Phenotypic patterns of antibiotic resistance of P. aeruginosa were not correlated with their prior antibiotic exposition. Fifteen of 18 patients (83.3%) had recurrent P. aeruginosa pneumonia caused by the strains with same PFGE pattern. Conclusion: These data suggest that the most of the recurrent P. aeruginosa infections in chronic lung disease occurred due to the relapse of prior infections. Further investigations should be performed for assessing the molecular mechanisms of the persistent colonization and for determining how to eradicate clonal persistence of P. aeruginosa.

• Asian-Australasian Journal of Animal Sciences
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• 제13권8호
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• pp.1040-1043
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• 2000

The genomic mapping of Red Jungle Fowl (Gallus gallus), local Village Chicken, and broiler was carried out by random amplified polymorphism DNA (RAPD) technique. Two different sets of arbitrary primers were used (Operon OPA01-20 and Genemed GM01-50). All the genomes of the three species of chickens were amplified with OPA01-20 primers. The genomes of the Red Jungle Fowl and local Village Chicken were further amplified with GM01-50 primers. Analysis of the results based on band sharing (BS) and the molecular size of individually amplified DNA fragments showed that Red Jungle Fowl and local Village Chicken shared the species similarity of 66% with Operon primers 01-20, 64% between local Village Chicken and broiler, and 63% when DNA bands between Red Jungle Fowl and broiler were compared. With GM01-50, the BS between Red Jungle Fowl and local village chicken increased to 72%. The results showed that the local village chicken is more closely related to Red Jungle Fowl than to broiler in the genetic distance. On the other hand, broiler is 1% closer in genetic distance to local village chicken than to Red Jungle Fowl. The results also indicated that primers like OPA-7, 8 and 9 can be used as species specific DNA markers for these three species of chickens.

• 한국약용작물학회지
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• 제21권4호
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• pp.247-254
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• 2013

Collected germplasms of five representative dandelion species (Taraxacum ohwianum, T. platycarpum, T. platypecidum, T. officinale, and T. coreanum) were 104 lines from different habitates in Korea and China. Their genetic diversity was analyzed by genomic fingerprinting method using amplified fragment length polymorphism (AFLP). AFLP results of 6 primer combinations were revealed 1,176 total DNA fragments and 523 polymorphic bands with a 44.4% ratio of polymorphism. On the basis of similarity coefficient analysis by unweight pair group method with arithmetic averages (UPGMA), 104 dandelion germplasm lines were ranged from 0.64 to 0.99 and clustered distinct five group depending on the species. Furthermore, a principal coordinate analysis (PCA) by the application of multi-variate analysis indicated significantly greater differences among species than geographical origins.

• 대한바이러스학회지
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• 제29권1호
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• pp.39-44
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• 1999

Warts, or verrucae, are benign epithelial proliferations of the skin and mucosa caused by infection with human papillomaviruses (HPV). It is now recognized that there are many different HPV types. Especially type3 is most frequently observed in flat wart. Other types, such as type2, 10, 14, 27, 28, 29, 38, and 41 are rarely encounted in flat wart. We describe here a simple and economic method for detection and identification of epidermodysplasia verruciformis-associated HPV. The method is based on polymerase chain reaction (PCR) amplification and restriction analysis. The method has been developed with cloned HPV DNA and DNA from clinical samples. Clinical samples are from either frozen tissue or paraffin-embedded tissue. Genomic fragments were obtained from two different HPV types (3 and 10). The amplification fragments were identified by a form of miniature fingerprinting, with a set of restriction enzymes that gave a unique digestion pattern for each HPV type. We have tested 74 clinical samples. Only type3 among these clinical samples is detected, and one sample is involved in neither type3 nor type10.

• 대한수의학회지
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• 제39권4호
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• pp.811-818
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• 1999

A total of 16 Staphylococcus epidermidis isolates collected from 14 dogs admitted to the Veterinary Medicial Teaching Hospital in Seoul National University over eleven months were examined for in vitro antibiotic susceptibility pattern with minimum inhibitory concentration (MIC) and slime production, a virulence-associated phenotype, and were genetically characterized by pulsed-field gel electrophoresis (PFGE). The frequency of resistance to antimicrobial agents tested was not high, with a susceptibility ranging from 56.3% to 100%. Three strains exhibited multiple drug resistance against amikacin (MIC, $32-64{\mu}g/ml$), ampicillin ($32{\mu}g/ml$), fosfomycin ($32-128{\mu}g/ml$) and gentamicin ($16{\mu}g/ml$). Vancomycin, ciprofloxacin and rifampin were effective antibiotics against the isolates. All isolates were slime producers ; strains isolated from dogs which died of bacteremia were more likely to produce slime than those isolated from dogs which survived. Chromosomal DNA fingerprinting of the isolates yielded 16 different genomic types with few common bands, indicating a variety of clones of S epidermidis were prevalent in the hospital. This study revealed that PFGE is an useful method for the genotype characterization of S epidermidis strains and this organism could probably be pathogenic in some dogs with severe disorders. Further works on a larger number of epidemiologically defined strains are required to assess these results.

• 한국약용작물학회지
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• 제21권6호
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• pp.455-460
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• 2013

Collected germplasms of five representative species belonging to Curcuma genus (C. longa, C. aromatica, C. zedoaria, C. phaeocaulis and C. kwangsiensis) were 52 samples from different farmhouse in Korea and China. To elucidate the genetic diversity among the species, 52 samples were analyzed by genomic fingerprinting method using amplified fragment length polymorphism (AFLP). AFLP results of 6 primer combinations were revealed 643 total DNA fragments and 349 polymorphic bands with the 54.3% ratio of polymorphism. In the analysis of coefficient similarity using unweight pair group method with arithmetic averages (UPGMA), 52 Curcuma germplasm lines were ranged from 0.60 to 0.99 and clustered distinct five groups according to the species and collected geographical levels. However, the result of principal coordinate analysis (PCA) by multi-variate analysis was shown significantly greater differences among species than geographical origins based on AFLP profiling data of these samples.

• Archives of Pharmacal Research
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• 제28권7호
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• pp.854-858
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• 2005

A Lactobacillus isolate was collected from the feces of a healthy Korean individual and named as Lactobacillus ruminus SPM0211. It was further characterized by subjecting it to an antibiotic resistance test and genetic analysis. In the antibiotic resistance test, all tested Lactobacillus spp. were classified as 'high resistance' for multiple antibiotics, such as isoniazid, ethambutol, cycloserine, and vancomycin. L. ruminus SPM0211 was classified as 'high resistance' for streptomycin also, while the other tested Lactobacillus spp. were classified as low resistance. This suggests that the antimicrobial spectra may be a good indicator in the discrimination of this strain among the tested Lactobacillus spp. In a polymerase chain reaction-random amplified polymorphic DNA (PCR-RAPD) analysis using the Microbial Uniprimer kit, L. ruminus SPM0211, and L. suebicus were clustered as a group with a 74.3% similarity level, suggesting that these two species are genetically related. Thus, our data suggest that the PCR-RADP method using the Microbial Uniprimer kit may be valuable in discriminating L. ruminus SPM0211 from other Lactobacillus spp.

• 치위생과학회지
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• 제19권2호
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• pp.86-95
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• 2019

The modern era of microbial genome analysis began in earnest in the 2000s with the generalization of metagenomics and gene sequencing techniques. Studying complex microbial community such as oral cavity and colon by a pure culture is considerably ineffective in terms of cost and time. Therefore, various techniques for genomic analysis have been developed to overcome the limitation of the culture method and to explore microbial communities existing in the natural environment at the gene level. Among these, DNA fingerprinting analysis and microarray chip have been used extensively; however, the most recent method of analysis is metagenomics. The study summarily examined the overview of metagenomics analysis techniques, as well as domestic and foreign studies on disease genomics and cluster analysis related to oral metagenome. The composition of oral bacteria also varies across different individuals, and it would become possible to analyze what change occurs in the human body depending on the activity of bacteria living in the oral cavity and what causality it has with diseases. Identification, isolation, metabolism, and presence of functional genes of microorganisms are being identified for correlation analysis based on oral microbial genome sequencing. For precise diagnosis and treatment of diseases based on microbiome, greater effort is needed for finding not only the causative microorganisms, but also indicators at gene level. Up to now, oral microbial studies have mostly involved metagenomics, but if metatranscriptomic, metaproteomic, and metabolomic approaches can be taken together for assessment of microbial genes and proteins that are expressed under specific conditions, then doing so can be more helpful for gaining comprehensive understanding.

• 한국축산식품학회지
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• 제22권4호
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• pp.301-309
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• 2002

본 연구는 AFLP-PCR 유전자 지문분석 기법을 이용하여 우리나라 고유의 동물유전자원으로서 재래흑염소의 품종 및 흑염소육 감별을 위한 품종 특이적 DNA marker를 개발하고자 수행하였다. 흑염소로부터 추출한 genomic DNA를 EcoR I/Hind III 및 Taq I/Hind III 2종류의 제한효소 조합으로 이중 절단한 후 10종류의 two selective primer조합형을 이용하여 분석한 결과 각 printer 조합형당 검출된 AFLP band의 수는 36~74개의 범위로 평균 55.5개였다. 그리고 검출된 총 555개의 band 가운데 polymorphic band의 수는 149 개로 다형성 수준은 약 26.8%로 추정되었다. 재래흑염소 품종 특이적인 AFLP marker를 탐색하고자 육용종 수입흑염소 및 4품종의 유용종 염소와 AFLP 지문양상을 비교 검토한 결과 M13/H13 primer 조합형에서 2.01과 1.26 kb의 2개 band 그리고 E35/H14 primer 조합형에서 1.65 kb의 1개 band가 재래흑염소의 품종 특이적 AFLP marker로 검출되었다. 그리고 E35/H14 primer 조합형에서 수입흑염소의 2.19, 2.03, 0.96 및 0.87 kb band, Saanen종의 2.13 kb band, Nubian종의 2.08 kb band는 각 해당 품종에만 특이적으로 출현하는 품종 특이적 band로 확인되었다. 또한, E35/H13 primer 조합형에서 재래흑염소를 특히, Saanen종과 식별이 가능한 4개의 DNA band가 확인되었다. 따라서, 본 연구에서 AFLP-PCR 기법을 이용하여 검출한 품종 특이적 DNA band들은 우리나라 재래흑염소, 수입흑염소 및 유용종 염소품종들간에 명확히 구별되어 재래종 흑염소 육과 육제품의 품종판별에 매우 유용한 DNA marker로 이용 가능할 것으로 기대된다.