• 미생물학회지
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• 제37권1호
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• pp.72-79
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• 2001

폐수 처리장의 방류수에 페놀을 첨가한 후 terminal restriction fragment length polymorphism (T-RFLP) 방법을 이용하여 세균군집의 구조와 변화를 조사하였다. 시료로부터 얻은 16S rRNA gene은 eubacterial primer로 증폭하였으며, 한 primer는 5'말단에 biotin을 부착하였다. 증폭된 product는 HaeIII와 AluI으로 각각 절단하였고, 절단된 단편 중에서 terminal restriction fragment (T-RF)를 streptavidin paramagnetic particle을 이용하여 분리하였다. 분리된 T-RF는 전기영동과 silver staining을 통하여 확인하였다. 본 실험의 유용성을 검증하기 위하여 표준 균주 10 균주를 대상으로 실험하였고, 균주마다 특징적인 T-RF를 가지는 것과 그 크기가 Ribosomal database project (RDP) 자료로부터 계산된 결과와 일치하는 것을 확인할 수 있었다. 한편, 대조군으로 사용된 페놀을 첨가하지 않은 방류수 시료에서는 Acinetobacter, Bacillus, Pseudomonas 속 등이 우점종을 차지하고 있었고, 페놀 (최종농도 250mg.$l^{-1}$)을 첨가한 방류수 시료에서는 Acinetobacter, Comamonas, Cytophaga, Pseudomonas 속 등이 우점종을 차지함을 알 수 있었다. Gel에서 분리한 Acinetobacter와 Cytophaga에 해당되는 T-RF는 재증폭 및 염기 서열 분석이 가능하였는데, database의 염기서열과 비교한 결과 Acinetobacter junii와 유연관계가 가깝다는 것을 확인하였다.

• 셀메드
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• 제10권3호
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• pp.19.1-19.6
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• 2020

The kidneys are exposed to toxicants and waste product and can be affected easily by these toxicants and by products of the metabolism. The consumption of adequate water is necessary to remove waste and to keep kidney healthy. Deficiency of liquid in the blood leads to various adverse effects on the kidney. The most common adverse deficiency of liquid in blood is deposition of solid matter in the kidney and subsequently formation of kidney stone. Nephrolithiasis (kidney stone) can be treated by drugs if it is small in size but if it blocks the route due to its big size then surgery is the only way to remove it. The recurrence rate of the problem is very high and it may reappear within 10 years. In Unani literature Hasāh wa Raml al-Kulya (nephrolithiasis) is described in detail. As per Unani literature stagnation of Ghalīz mādda (filthy and viscous matter) in the kidney is the main cause of the formation of kidney stone. Various single and compound formulations drugs are described for the management of kidney stone which are very effective as well as safe. Management is divided into two parts i.e. symptomatic treatment to relieve pain and to methods adopted to remove stone from the kidney. Musakkin-i-Waja'(analgesic) drugs are used for pain while Mufattit-i-Hasāh (lithotriptic) and Mudirr-i-Bawl (Diuretic) drugs are used to remove stone. Majoon Aqrab, Qurs Kaknaj and Dawa-e-Gurda etc. are compound drugs mentioned in literature for removal of kidney stone. Single drugs like Alu Balu, Tukhm Khayar, and Kharkhask etc. are also used for same purpose.

• The Plant Pathology Journal
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• 제23권1호
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• pp.13-21
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• 2007

Sweet potato feathery mottle virus(SPFMV) is one of the most prevalent viruses infecting sweet potatoes and occurs widely in sweet potato cultivating areas in Korea. To assess their genetic variation, a total of 28 samples infected with SPFMV were subjected to restriction fragment length polymorphism(RFLP) analysis using DNAs amplified by RT-PCR with specific primer sets corresponding to the coat protein(CP) region of the virus. The similarity matrix by UPGMA procedure indicated that 28 samples infected with SPFMV were classified into three groups based on the number and size of DNA fragments by digestion of CP-encoding regions with 7 enzymes including SalI, AluI, EcoRI, HindIII, FokI, Sau3AI, and DraI bands. Four primer combinations out of 5 designed sets were able to differentiate SPFMV and sweet potato virus G infection, suggesting that these specific primers could be used to differentiate inter-groups of SPFMV. Sequence analysis of the CP genes of 17 SPFMV samples were 97-99% and 91-93% identical at the intra-group and inter-groups of SPFMV, respectively. The N-terminal region of the CP is highly variable and examination of the multiple alignments of amino acid sequences revealed two residues(residues 31 and 32) that were consistently different between SPFMV-O and SPFMV-RC.

• 미생물학회지
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• 제41권1호
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• pp.24-37
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• 2005

국내 사육 하우 염소, 돼지, 개, 닭 및 마우스 등을 포함한 각종 동물과 사람환자에서 분리한 총 116주의 S. aureus 분리주에 대해서 5종의 PCR 기법을 적용하여 분자역학적 특성을 분석하였다. 먼저 종특이 유전자(SSG: aroA, coa, nuc 및 spa-gene)의 다양성을 PCR 기법으로 조사하였고, 또한 약제내성의 MRSA 균주의 분포양상과 내독소(Enterotoxin)산생유전자(SE)의 분포양상에 대해서도 조사하였다. 그리고 이 연궁선 수행한 5종의 PCR 기법 들이 생산한 개별성적을 객관적으로 비교하여, 가장 신뢰도 높고 효율적 PCR기법을 선발하였다. 먼저 PCR기법을 적용한 SSG의 분포양상 조사에서는 $nuc-gene\;(100\%)$, $spa-gene\;(91.4\%)$, $coa-gene\;(87.9\%)$, 및 $aroA-gene\;(26.7\%)$의 빈도로 조사되었다. 그리고 aroA와 coa-gene PCR 증폭산물에 대한 RsaI과 AluI 효소로 소화시킨 RELP 성적에서 coa-Bene PCR-RFLP에서는 모두 10 type의 con-type이 동정되었고, 그중 coa-3 type (809 bp)이 닭, 마우스, MRSA 및 사람유래 균주를 포함한 총 36주 $(33.0\%)$로서 가장 대표적인 genotype으로 결정되었다. 반면에 aroA-gene PCR-RELP에서는 단지 7 type의 genotype만이 산생되어 대조를 보였고, 17주 $(73.9\%)$가 대표적인 그룹으로 분류되었다. 한편 spa-gene PCR에서는 총 11 type의 spa-type이 확인되었고, 그중 spa-7 type (9 repeats; 263 bp)이 한우, 닭, 개, 염소, 마우스 및 MRSA 균주 등을 포함한 총 39주 $(34.8\%)$로서 가장 대표적인 genotype으로 결정되었다. 또한 총 116주에 대해 MRSA 균주의신속한 검출을 위해서 mecA-gene PCR을 적용하였던 바, 단지 사람유래의 14주 $(12.1\%)$에서만 양성의 증폭산물 (533 bp)이 검출되었고, 이들 증폭된 PCR산물의 유전학적 검증을 위해 HhaI으로 소화시켰던바 14주 모두 $(100\%)$가 알려진 2개의 DNA band (332 & 201 bp)로 양분되어 유전자수준에서 MRSA 양성균주로 검증되었다. 한편 내독소 산생유전자 (SE-gene)는 multiplex-PCR기법으로 조사하였으며 sea-gene $(63.7\%)$이 가장 지배적이었고, 이어서 seb-gene $(10.0\%)$ 및 sec-gene $(7.2\%)$의 순서였다. 특히 sea+sec의 2종의 SE genes를 보유한 균주도 11주로서 가장 많았으며 그 외에도 sea+seb (2주)및 seb+see (1주) genes를 보유한 균주도 함께 검출되었다. 최종적으로 5종의 PCR기법들이 생산한 성적들을 수치 (SID)로 변환하여 객관적으로 감별능력 (DA)을 비교하였던바, aroA-gene PCR-RFLP는 가장 저조한 DA [SID=0.462]을 나타내었고, 반면에 coa-gene PCR-RELP는 5종의 분석기법 중에서 가장 신뢰도 높은 DA [SID=0.894]을 발현하였다. 따라서 현재의 연구결과con-gene PCR-RELP기법이 사람을 포함한 다양한 동물종유래 S. aureus 균주들의 유전학적 분석목적에 가장 신뢰도 높고 감별능력이 뛰어난 분석기법으로 선발되었다.

• The Plant Pathology Journal
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• 제33권2호
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• pp.140-147
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• 2017

Fusarium wilts of strawberry, caused by Fusarium oxysporum f. sp. fragariae, is a serious soil-borne disease. Fusarium wilt causes dramatic yield losses in commercial strawberry production and it is a very stubborn disease to control. Reliable chemical control of strawberry Fusarium wilt disease is not yet available. Moreover, other well-known F. oxysporum have different genetic information from F. oxysporum f. sp. fragariae. This analysis investigates the genetic diversity of strawberry Fusairum wilt pathogen. In total, 110 pathogens were isolated from three major strawberry production regions, namely Sukok, Hadong, Sancheong in Gyeongnam province in South Korea. The isolates were confirmed using F. oxysporum f. sp. fragariae species-specific primer sets. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses were executed using the internal transcribed spacer, intergenic spacer, translation elongation factor1-${\alpha}$, and ${\beta}$-tubulin genes of the pathogens and four restriction enzymes: AluI, HhaI, HinP1I and HpyCH4V. Regarding results, there were diverse patterns in the three gene regions except for the ${\beta}$-tubulin gene region. Correlation analysis of strawberry cultivation region, cultivation method, variety, and phenotype of isolated pathogen, confirmed that genetic diversity depended on the classification of the cultivated region.

• Journal of Microbiology and Biotechnology
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• 제21권10호
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• pp.1001-1011
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• 2011

To explore bacterial diversity for elucidating genetic variability in acylhomoserine lactone (AHL) lactonase structure, we screened 800 bacterial strains. It revealed the presence of a quorum quenching (QQ) AHL-lactonase gene (aiiA) in 42 strains. These 42 strains were identified using rrs (16S rDNA) sequencing as Bacillus strains, predominantly B. cereus. An in silico restriction endonuclease (RE) digestion of 22 AHL lactonase gene (aiiA) sequences (from NCBI database) belonging to 9 different genera, along with 42 aiiA gene sequences from different Bacillus spp. (isolated here) with 14 type II REs, revealed distinct patterns of fragments (nucleotide length and order) with four REs; AluI, DpnII, RsaI, and Tru9I. Our study reflects on the biodiversity of aiiA among Bacillus species. Bacillus sp. strain MBG11 with polymorphism (115Alanine > Valine) may confer increased stability to AHL lactonase, and can be a potential candidate for heterologous expression and mass production. Microbes with ability to produce AHL-lactonases degrade quorum sensing signals such as AHL by opening of the lactone ring. The naturally occurring diversity of QQ molecules provides opportunities to use them for preventing bacterial infections, spoilage of food, and bioremediation.

• 대한미생물학회지
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• 제35권1호
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• pp.69-76
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• 2000

Members of the genus Acinetobacter are recognized as newer pathogens of the nosocomial infection with an increasing frequency in recent years. Strains that belonged to A. calcoaceticus A. baumannii complex (genomic species 1, 2, 3, and 13TU) were major groups associated with nosocomial infection. Phenotypic identification was unreliable and laborious method to classify Acinetobacter strains into 19 genomic species. Rapid and reliable identification of clinical isolates is essential to diagnosis and epidemiology of Acinetobacter. We investigated the suitability of amplified ribosomal DNA restriction analysis (ARDRA) to identify genomic species of 131 Acinetobacter isolates. The 16S rRNA genes (ribosomal DNA) were enzymatically amplified and the amplified PCR products were restricted independently with the enzymes, AluI, CfoI, and MboI. Genomic species of Acinetobacter was classified by the combinations of restriction patterns. The analysis was showed that restriction profiles were characteristic for each genomic species. One hundred fourteen isolates were identified as A. baumannii, twelve were identified as genomic species 13TU, and one was identified as genomic species 3. Four isolates were found to be unknown organisms. All of the isolates which were identified to A. baumannii by phenotypic tests were completely discriminated into A. baumannii and genomic species 13TU by ARDRA. This study demonstrates that ARDRA is a rapid and simple techniques for the identification of Acinetobacter species according to the genomic species.

• 한국식물병리학회지
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• 제13권4호
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• pp.239-243
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• 1997

1990년 8월 충북 보은의 참깨 밭에서 전형적인 엽화병(葉化病; phyllody disease)이 발생하였다. 병징은 꽃이 잎처럼 녹색으로 변하고, 이병엽은 정상엽보다 작고 잔가지가 마치 빗자루 모양으로 총생하였고 개화 및 결실이 되지 않았다. 투과 전자현미경으로 병든 잎의 사관요소에서 phytoplasma가 존재하는 것을 확인하였다. 병든 잔가지에서 추출한 DNA를 중합효소 연쇄반응으로 분석한 결과 대추나무 빗자루병에 걸린 나무에서 증폭되는 DNA와 같은 크기의 band가 관찰되었다. 따라서 위의 시료는 phytoplasma 벙으로 확인 되었으며 참깨 엽화병으로 명명하였다. 또한 참깨 엽화병을 일으키는 phytoplasma와 대추 나무 빗자루병 phytoplasma를 PCR 증폭 및 제한효소로 절단하여 분석한 결과 이들은 서로 다른 그룹이라는 것을 알 수 있었다.

• Food Science and Biotechnology
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• 제17권4호
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• pp.888-891
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• 2008

Twenty-three fungal strains were isolated from meju that had originated from the Sunchang province, the famous location for making fermented soybean foods in Korea. The restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS) region of the rDNA (ITS-RFLP) was applied to differentiate the isolated fungal strains. First, the ITS region by polymerase chain reaction (PCR) with specific primers was amplified and then cleaved the products with different restriction enzymes. Cleavage of the amplified fragments with the restriction enzymes AluI, HaeIII, HhaI, and TaqI revealed extensive polymorphisms. The ITS-RFLP results highly correlated with ITS sequence analysis. All of the 23 fungal strains were classified into 5 groups by ITS-RFLP analysis. Aspergillus oryzae was the major fungal strain isolated from Sunchang meju (12 out of 23), while Aspergillus fumigatus was the next most frequently isolated strain (7 out of 23). In contrast, it was found that Fusarium asiaticum, Aspergillus sydowii, and Arthrinium sp. were the minor fungal strains in meju.

• Asian-Australasian Journal of Animal Sciences
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• 제17권10호
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• pp.1334-1337
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• 2004

The present study was conducted to detect polymorphism at growth hormone gene in Karan Fries bulls. A 428 bp fragment of growth hormone gene spanning over $4^{th}$exon, $4^{th}$intron and $5^{th}$ exon was amplified and digested with AluI restriction enzyme to identify polymorphism at this locus. Karan Fries bulls were found to be polymorphic at this locus. Two genotypes LL and LV were identified in Karan Fries with higher allelic frequency for L allele. In Karan Fries males, the average birth weight, 3 months body weight and daily body weight gains of LL homozygotes were significantly higher than that of LV heterozygotes. Genetic distances of KF bulls with respect to genotype along with 3 months body weight and average daily body weight gain forms a single cluster of bulls with LL genotype, while individuals with LV genotype forms three distinct clusters indicating more influence of L allele on growth traits.