• Korean Chemical Engineering Research
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• 제48권2호
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• pp.147-156
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• 2010

폐가스 처리용 바이오필터의 핵심 요소 기술은 생촉매(미생물), 담체, 설계 운전 기술 및 진단 관리 기술이다. 특히, 바이오필터의 성능은 부하 조건과 바이오필터 내 미생물 군집 구조에 의해 영향을 받는다. 지금까지 바이오필터의 미생물 연구는 대부분 배양법을 기초로 하여 수행되어 왔으나, 최근에 보다 신속하고 정확하게 미생물 군집을 분석할 수 있는 방법들이 제시되고 있다. 본 논문에서는 생리적, 생화학적 및 분자생물학적 미생물 군집 분석 방법과 이를 활용한 바이오필터의 미생물 군집 특성을 조사한 연구사례를 소개하고, 미생물 군집 분석법의 바이오필터에 적용 가능성에 대해 고찰하였다. Community-level physiological profile 방법은 시료 중에 포함된 종속영양미생물의 탄소기질 이용능력을 기반으로 군집 특성을 조사하는 것이며, Phospholipid fatty acid analysis는 미생물 세포막 지방산을 분석하여 군집 특성을 조사하는 방법이다. 환경시료로부터 직접 추출한 DNA를 활용하는 분자생물학적 분석법에는 "partial community DNA analysis"와 "whole community DNA analysis"가 있다. 전자의 방법은 PCR 과정에 의해 증폭시킨 염기서열을 분석하는 것으로 ribosomal operon 유전자가 가장 많이 활용되었다. 이 방법은 다시 PCR fragment cloning 및 genetic fingerprinting으로 구분되며, genetic fingerprinting 방법으로는 denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis 및 random amplified polymorphic DNA 방법으로 세분화된다. 추출된 전체 군집의 DNA를 분석하는 방법에는 total genomic cross-DNA hybridization, 총 추출 DNA의 열 변성/재결합 방법 및 밀도구배를 이용하여 추출한 DNA를 분획화하는 방법 등이 있다.

• 한국동물위생학회지
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• 제32권3호
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• pp.257-264
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• 2009

Subtyping of Brucella abortus isolates is epidemiologically important for monitoring of bovine brucellosis outbreaks. Pulsed-field gel electrophoresis (PFGE) is considered as a gold standard of molecular typing methods to study the DNA polymorphisms of bacteria. In this study, we analyzed using PFGE the DNA fragment profiles of B. abortus isolated in Gyeongbuk province from 1998 to 2006. The genomic DNA was digested with the restriction endonuclease Xba I, Xho I and Smi I followed gel electrophoresis. No distinguishable patterns of the genomic DNA digested with Xba I and Xho I were observed among the field isolates of B. abortus tested in this study. But Smi I restriction enzyme resulted in two PFGE patterns consisting of 13-15 bands that ranged in size from 33 to 668bp by standard marker. The cluster analysis by DNA fingerprinting software showed 93.75% similarity between two PFGE patterns. No different PFGE patterns were recognized among the isolates originated from various years, regions and cow breeds.

• 한국한의학연구원논문집
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• 제1권1호
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• pp.471-476
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• 1995

Natural products used for oriental medicine often come from various geographical sources, after several different distribution channels. Therefore some form of quality control procedure is required to safeguard naturl products for prescriptions purposes. To achieve this, systematic apprroaches such as morphological examination, microscopic analysis of powdered herbs and chemical analysis can be carried out. However, to ensure absolute criteria for quality assurance of natural products, DNA fingerprinting method such as RAPD(Random amplified polymorphism DNA) analysis can be used for authentication of natural products for authenticatin of natural products. In this study, warious oligonucleotide primers will be synthesized for the detection of RAPD markers and also parameters of affecting PCR(Polymerase Chain Reaction) in the detection of RAPD markers of rare and high-priced natural products will be studied with genomic DNA of chosen samples.

• 대한미생물학회지
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• 제35권5_6호
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• pp.349-349
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• 2000

• 대한의생명과학회지
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• 제9권4호
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• pp.183-187
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• 2003

Pseudomonas aerugionsa is a commonly isolated nosocomial pathogen. DNA fingerprinting of P. aerugionsa is examined by randomly amplified polymorphic DNA (RAPD). In this study, P. aeruginosa were isolated from environmental and clinical specimens and the molecular typing of the microorganisms was investigated by RAPD. Thirty strains of P. aeruginosa were selected from the strains isolated formerly and submitted for type identification to the University Hospital. 15 strains of P. aeruginosa were received from Chungnam University Hospital and 14 strains from Gyeongsang University Hospital. DNA of P. aeruginosa was extracted by Qiagen genomic DNA kit. PCR mixtures were set up and incubated, Reactions mixtures were made to be optimal for P. aeruginosa. RAPD typing analysis was carried out by the multivariate statistical program (MVSP) V3.0. RAPD type I was the most common pattern and included 23 strains. Most of strains from Gyeongsang University Hospital belonged to RAPD type lb and 15 strains from Chungnam University Hospital to RAPD type I or II. RAPD typing of P. aeruginosa isolated from the environmental and clinical specimens was very simple and reproducible.

• 대한의생명과학회지
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• 제11권3호
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• pp.295-299
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• 2005

Staphylococcus aureus is an important animal and human pathogen implicated in a variety of disease including food-poisoning caused by staphyloccal enterotoxins (SEs). In order to investigate the difference in genomic types and to monitor the transmission of S. aureus isolates, a total of 25 S. aureus isolates from different sources were determined for their genotypic characteristics by pulsed-field gel electrophoresis (PFGE) in addition to their ability to enterotoxin production and antibiotic resistance patterns in this study. All the isolates were susceptible to amikacin, and the resistance pattern to ampicillin and penicillin were most common among 14 different patterns. Eleven of 24 isolates produced one of three SEs, SEA, SEC or SED. Sixteen representative PFGE patterns were obtained by Smal restriction fragments of S. aureus isolates. Analysis of dendrogram based on PFGE band patterns suggested that food-poisoning outbreaks be caused by the diverse sources of food, of which their raw materials were infected with S. aureus. Also, it could be concluded that PFGE was a powerful tool for epidemiological tracing of infection source for food-initiated outbreaks.

• Journal of Microbiology and Biotechnology
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• 제21권12호
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• pp.1336-1344
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• 2011

This study details and examines a novel ligation-mediated polymerase chain reaction (LM-PCR) method. Named the LM-PCR/Shifter, it relies on the use of a Class IIS restriction enzyme giving restriction fragments with different 4-base, 5' overhangs, this being the Shifter, and the ligation of appropriate oligonucleotide adapters. A sequence of 4-base, 5' overhangs of the adapter and a 4-base sequence of the 3' end of the primer(s) determine a subset of the genomic restriction fragments, which are amplified by PCR. The method permits the differentiation of bacterial species strains on the basis of the different DNA band patterns obtained after electrophoresis in polyacrylamide gels stained with ethidium bromide and visualized in UV light. The usefulness of the LM-PCR/Shifter method for genotyping is analyzed by a comparison with the restriction endonuclease analysis of chromosomal DNA by the pulsed-field gel electrophoresis (REA-PFGE) and PCR melting profile (PCR MP) methods for isolates of clinical origin. The clustering of the LM-PCR/Shifter fingerprinting data matched those of the REA-PFGE and PCR MP methods. We found that the LM-PCR/Shifter is rapid, and offers good discriminatory power and excellent reproducibility, making it a method that may be effectively applied in epidemiological studies.

• 대한수의학회지
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• 제41권2호
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• pp.157-165
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• 2001

Amplified fragment length polymorphism(AFLP) technique is based on the polymorphism detection through selective PCR amplification of restriction fragments from digested genomic DNA and thus includes the procedures of the total DNA digestion by endonucleases, ligation of adapters to the ends of the fragments, and following the selective amplification of the restricted DNA fragments. This study were aimed to : (1) determine the genetic variability of S aureus strains, (2) estimate genetic diversity within and among these strains, (3) compare phylogenetic relationships among these strains as genetic markers using AFLP techniques. Genomic DNA was digested with a particular combination of three restriction enzymes with specific recognition sites and the DNA fragments were ligated to restriction specific adapters and amplified using the selective primer combinations. In the S aureus strain, the number of scorable AFLP bands detected per each primer combination varied from 29 to 102, with an average of 61.59 using 27 primer combinations. A total of 1,663 markers were generated, 904 bands of which were polymorphic, showing a 33.48% level of polymorphism with these primer combinations. Among the primer combinations, E02/T02, E02/T03, E04/H02, E02/T01 and E04/H03 primer combinations showed a high level of polymorphism with 0.78, 0.76, 0.74, 0.71 and 0.70, respectively. But T03/H01, E01/T02 and E01/T03 primer combinations showed a low level of polymorphism with 0.38, 0.37 and 0.15, respectively, Therefore, the former primer combinations will be the most effective for AFLP analysis of S aureus. In SA1 sub-types the level of polymorphism of S aureus KCTC 1927 was similar to that of S aureus CU 01(0.825) and higher than those of other strains such as S aureus CU 02 (0.715), S aureus KCTC 2199(0.625), S aureus KCTC 1916(0.607) and S aureus KCTC 1621 (0.553). In SA2 sub-types the level of polymorphism of S aureus CU 07 was similar to that of S aureus CU 08(0.935) and higher than those of both S aureus CU 04(0.883) and S aureus CU 05(0.883) and lower than those of S aureus CU 03(0.583). In SA3 subtypes the level of polymorphism of S aureus CU 11 was similar to that of S aureus CU 12(0.913) and lower than that of S aureus CU 15(0.623). The results proved that AFLP marker analysis of S aureus strain could be used to study the epidemiology of mastitis and in addition, common genotype in geographic region could be useful for the development of an effective vaccine or DNA marker for easy diagnosis of mastitis caused by S aureus infection.

• Journal of Periodontal and Implant Science
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• 제25권1호
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• pp.89-98
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• 1995

P. intermedia are considered an important pathogen in adult periodontitis, rapidly progressing periodontitis, refractory periodontitis, pregnancy gingivitis, acute necrotizing ulcerative gingivitis, pubertal gingivitis. So far 2 DNA homology groups and 3 serotypes of P. intermedia have been reported but there is no data available as yet regarding genetic diversity for the species P. intermedia. The purpose of this study is to investigate, using bacterial DNA restriction endonuclease analysis, genetic diversity between individual strains of P. intermedia which are indistinguishable by serotyping and biotyping, occurrence of an intrafamilial transmission and genetic heterogeneity between P. intermedia strains isolated within a patient and within the same serotypes. The families who have had no systemic disease, no experience of periodontal treatment for the previous 1 year and no experience of antibiotics for the previous 6 months were selected and subgingival plaque was collected at 4 sites in each person and incubated in the anaerobic chamber. P. intermedia were identified by colony shape, gram stain, biochemical test, SK-I03(Sunstar Inc.) test and IIF using monoclonal antibody was perfomed for the determination of serotypes. P. intermedia strains were grown in BHI broth and whole genomic DNA was extracted and digested by restriction endonuclease. The resulting DNA fragments were separated by agarose gel electrophoresis, stained and photographed under UV. As the results of this study, intrafamilial vertial transmissions could be assessed in 2 families and horizintal transmissions in another 2 families. There were different DNA digest patterns within a patient, so P. intermedia showed that individuals could be colonized by multiple clonal types at anyone time. And different serotypes could be found within a patient and in the same serotype within a patient, obvius genetic heterogeneity could not be assessed. But in the same serotype in different famies, there were differences in the DNA digest patterns.

• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.1027-1031
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• 2010

To examine the possibility of horizontal gene transfer between transgenic potatoes and microorganisms in potato fields, the gene flow from transgenic potatoes containing the nucleoside diphosphate kinase 2 (NDPK2) gene to microorganisms in soils was investigated. The soil samples collected from the potato fields from March to October 2007 were examined by PCR, Southern hybridization, and AFLP fingerprinting. The NDPK2 gene from soil genomic DNAs was not detected by both PCR and Southern hybridization, indicating that gene transfer did not occur in the potato fields. In addition, no discrepancy was found in pathogenicity and noticeable changes for the appearance of variants of Phytophthora infestans in each generation when serial inoculations and the analysis of genomic DNAs by AFLP were conducted. Thus, these data suggest that transgenic potatoes do not give significant impacts on the communities of soil microorganisms and the emergence of variants, although continued research efforts may be necessary to make a decisive conclusion.