• Parasites, Hosts and Diseases
• /
• 제46권3호
• /
• pp.157-164
• /
• 2008

Three Acanthamoeba isolates (KA/E9, KA/E17, and KA/E23) from patients with keratitis were identified as Acanthamoeba triangularis by analysis of their molecular characteristics, a species not previously recognized to be a corneal pathogen. Epidemiologic significance of A. triangularis as a keratopathogen in Korea has been discussed. Morphologic features of Acanthamoeba cysts were examined under a microscope with differential interference contrast (DIC) optics. Mitochondrial DNA (mtDNA) of the ocular isolates KA/E9, KA/E17, and KA/E23 were digested with restriction enzymes, and the restriction patterns were compared with those of reference strains. Complete nuclear 188 and mitochondrial (mt) 16S rDNA sequences were subjected to phylogenetic analysis and species identification. mtDNA RFLP of 3 isolates showed very similar patterns to those of SH621, the type strain of A. triangularis. 16S and 18S rDNA sequence analysis confirmed 3 isolates to be A. triangularis. 18S rDNA sequence differences of the isolates were 1.3% to 1.6% and those of 16S rDNA, 0.4% to 0.9% from A. triangularis SH621. To the best of our knowledge, this is the first report, confirmed by 18S and 16S rDNA sequence analysis, of keratitis caused by A. triangularis of which the type strain was isolated from human feces. Six isolates of A. triangularis had been reported from contaminated contact lens cases in southeastern Korea.

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

• Journal of Microbiology
• /
• 제43권1호
• /
• pp.1-10
• /
• 2005

We isolated and cultured bacteria inhabiting solar saltern ponds in Taean-Gun, Chungnam Province, Korea. All of the isolated 64 strains were found to be moderately halophilic bacteria, growing in a salt range of 2-20 %, with an optimal concentration of 5% salt. Bacterial diversity among the isolated halophiles was evaluated via RFLP analyses of PCR-amplified 16S rDNAs, followed by phylogenetic analysis of the partial 16S rDNA sequences. The combination of restriction enzyme digestions with HaeIII, CfoI, MspI and RsaI generated 54 distinct patterns. A neighbor-joining tree of the partial 16S rDNA sequences resulted in the division of the 64 strains into 2 major groups, 45 strains of ${\gamma}-Proteobacteria$ (70.3%) and 19 strains of Firmicutes (29.7%). The ${\alpha}-Proteobacteria$ and Cytophaga-Flavobacterium-Bacterioides groups, which were repeatedly found to exist in thalassohaline environments, were not represented in our isolates. The ${\gamma}-Proteobacteria$ group consisted of several subgroups of the Vibrionaceae (37.5%), Pseudoalteromonadaceae (10.9%), Halomonadaceae (7.8%), Alteromonadaceae (7.8%), and Idiomarinaceae (6.3%). Members of Salinivibrio costicola (29.7%) were the most predominant species among all of the isolates, followed by Halobacillus treperi (12.5%). Additionally, three new species candidates were found, based on similarities of the 16S rDNA sequences to those of previously published species.

• The Plant Pathology Journal
• /
• 제35권1호
• /
• pp.51-62
• /
• 2019

A great variable response was observed when PP-3 and PP-J encumbered with 116 populations of root knot nematode (RKN) at two different temperatures ($25{\pm}2^{\circ}C$ and $30{\pm}2^{\circ}C$) and concentrations ($10^4$ and $10^5$ spores/ml). The PCR reaction amplified intergenic region between cytochrome oxidase subunit II gene (COII) and large subunit of rRNA gene (lrRNA) of the mitochondrial genome of different RKN species. The primer C2F3 and 1108 identified M. incognita with the highest frequency (52.6%) followed by M. javanica (36.8%) and M. arenaria (10.5%). The sizes of PCR products were 1.7 kb for M. incognita and M. javanica populations while populations of M. arenaria produced 1.1 kb fragment. The digestion with Hinf I yielded three different fragment length patterns on 1.5 % agarose gel. From current research it is concluded that intra-Meloidogyne genetic variability exist in RKN populations which have better encumbrance with P. penetrans.

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
• /
• pp.147.2-148
• /
• 2003

Complete nucleotide sequence of genome RNA of a Korean isolate of Pepper mottle virus (PepMoV-Vb) from field-collected diseased paprika (Capsicum annuum var grossum) was determined in this study. Symptoms of isolates of PepMoV were divided largely into two groups, vein banding (Vb) and vein clearing (Vc) patterns. PepMoV-Vb RNA consists of 9,640 nucleotides excluding the poly(A) tail. A single open reading frame was identified beginning at nucleotide position 169 encoding a polyprotein of 3024 amino acids which is typical of the Potyvirus genus. The complete nucleotide sequence and coding regions of PepMoV-Vb were compared to that of 11 potyviruses within the genus Potyvirus. The overall nucleotide sequence identity was 94.7 and 94.1％ identical to PepMoV-C and PepMoV-FL, respectively. Full-length cDNAs of PepMoV-Vbl were synthesized from purified viral RNAs by RT-PCR and their genome structure was analysed by RFLP analysis. This is the first report on complete nucleotide sequence of PepMoV isolated from paprika in Korea.

• Asian-Australasian Journal of Animal Sciences
• /
• 제14권7호
• /
• pp.915-921
• /
• 2001

IGF-I is involved in the regulation of growth and differentiation in mammals, but its role as a modulator of growth and metabolism in poultry is poorly understood. And, no studies have so far been reported for the comparison between serum IGF-I concentration and body growth in the egg type or the dual purposes (meat and egg type) chicken including the Korean Native Ogol Chicken (KNOC). Therefore, in order to improve the body growth and meat production of the KNOC, this study was conducted for the identification of the polymorphism of IGF-I gene and for its possible association with both body weight and IGF-I concentration. The RFLP patterns for IGF-I gene were identified by the PstI restriction enzyme. The frequencies of +/+, +/-, and -/- genotype were 16.9%, 51.7%, and 31.4%, respectively. Any statistical significance was not observed in all variations except for sex variation (p<0.01) by covariate quadratic model. The significant effect of the IGF-I genotype on body weight by sex indicates that there are different physiological characteristics in gender. Although the body weights of male KNOCs in most ages were not significant, there was a tendency of KNOCs with +/+ IGF-I genotype to be heavier than those with any other genotypes. But all IGF-I genotypes in female did not influence on body weight. The ANOYA revealed no significant effects of IGF-I genotypes on serum IGF-I concentration but sex effect was highly significant on the IGF-I concentration at 20 and 40 weeks (p<0.01). Although the +/+ genotype, in gender, tended to express a higher IGF-I concentration than the other genotypes at all ages in males, a statistical difference among the genotypes was not found except for 60 weeks (p<0.05). Furthermore, since body weight and IGF-I genotypes are associated, it is possible to improve KNOC to a meat type breed if a continuous selection can be made for the body weight and/or IGF-I traits.

• Parasites, Hosts and Diseases
• /
• 제35권4호
• /
• pp.277-282
• /
• 1997

만손열두조충과 북미열두조충 (Spirometra mansonoides)의 형태 차이점은 성숙편절의 자궁의 형대 가 전자근 차곡차곡 쌓인 꼴이고 후자는 알과벳 씨자 (C)형태이라는 점이다 이 비슷한 명태의 두 종 의 조충이 유전학적으로는 얼마나 차이가 있는지를 알기 위하여 중합효소연쇄반응-마디길이여러꼴 분석법(polymerase chain reaction-restriction fragment length polymorphism analysis)을 이용하여 유전 형질을 비교하였다. 충체로부터 285리넓솜 리보핵산 (285 rDNA), 사립체 cytochlmsc 산화 효소 아단위 I (mitochondrial cytochrome c oxidase subunit 1, mCOI) 및 리보솜 내부 전사된 영역 1 (ribosomal internal transcribed spacer 1, ITSI)에 대한 중합효소반응 산물을 구하였다. 이 산를은 Msp I. Hue III, Alu I, Cfo I, Rsc I의 4 염기 제한 효소로 잘라서, 전기영동하여 길과를 PAUP 3.1.1을 이용하여 분석 하였다. 285 리보솜 리보핵산과 리보솜 내부 전사된 영역 1 유진자에서는 두 충체 가 동일 한 분류가지에 묶였고 홀로서기수 (bootstrap number)는 94, 100이었다. 사립체 cytochrome c 산 화효소 아단위 1에서는 다른 분류가지에 묶였고 홀로서기수가 74이었다. 위 결과로 투 조충이 같은 조상에서 유래함과 진화 단계에서 매우 가까운 위치에 있음을 알 수 있었다.

• Toxicological Research
• /
• 제32권1호
• /
• pp.81-87
• /
• 2016

Aflatoxin B1 (AFB1) is produced by Aspergillus flavus growing in feedstuffs. Early detection of maize contamination by aflatoxigenic fungi is advantageous since aflatoxins exert adverse health effects. In this study, we report the development of an optimized conventional PCR for AFB1 detection and a rapid, sensitive and simple screening Real-time PCR (qPCR) with SYBR Green and two pairs of primers targeting the aflR genes which involved aflatoxin biosynthesis. AFB1 contaminated maize samples were divided into three groups by the toxin concentration. Genomic DNA was extracted from those samples. The target genes for A. flavus were tested by conventional PCR and the PCR products were analyzed by electrophoresis. A conventional PCR was carried out as nested PCR to verify the gene amplicon sizes. PCR-RFLP patterns, obtained with Hinc II and Pvu II enzyme analysis showed the differences to distinguish aflatoxin-producing fungi. However, they are not quantitative and need a separation of the products on gel and their visualization under UV light. On the other hand, qPCR facilitates the monitoring of the reaction as it progresses. It does not require post-PCR handling, which reduces the risk of cross-contamination and handling errors. It results in a much faster throughout. We found that the optimal primer annealing temperature was $65^{\circ}C$. The optimized template and primer concentration were $1.5{\mu}L\;(50ng/{\mu}L)$ and $3{\mu}L\;(10{\mu}M/{\mu}L)$ respectively. SYBR Green qPCR of four genes demonstrated amplification curves and melting peaks for tub1, afIM, afIR, and afID genes are at $88.0^{\circ}C$, $87.5^{\circ}C$, $83.5^{\circ}C$, and $89.5^{\circ}C$ respectively. Consequently, it was found that the four primers had elevated annealing temperatures, nevertheless it is desirable since it enhances the DNA binding specificity of the dye. New qPCR protocol could be employed for the determination of aflatoxin content in feedstuff samples.

• 한국습지학회지
• /
• 제11권1호
• /
• pp.123-130
• /
• 2009

지표수-지하수 혼합대는 지표수와 지하수의 수리적 교환이 일어나는 하상의 간극수 지역으로 모든 자연하천에 존재한다. 이 지역은 그동안 다른 특성으로 분리되었던 지표수와 지하수의 전이대로 다양한 미생물이 서식하고 있으며, 하변과는 또 다른 생지화학적인 기작이 일어날 것으로 추정된다. 지금까지의 연구는 미미하지만, 지표수-지하수 혼합대를 고려한 하천 생태 연구는 하천 전체의 생지화학적 기작을 이해하는데 중요한 역할을 할 것으로 보인다. 본 연구 대상 지역은 내성천(경상북도 예천군 고평교 하류 구간)으로 이 하천은 사질 하상이며, 자연이 그대로 보존되어있다고 평가 받는 자연 직할 하천이다. 토양시료는 하변 달뿌리풀(Phragmites japonica) 식생지, 나지, 수변 혼합대와 사주 혼합대 하강류(downwelling), 상승류(upwelling) 지역에서 깊이 별로 채취하여 수분함량, pH 및 용존유기탄소와 같은 토양 이화학적인 분석과 체외 미생물 효소($\beta$-glucosidase, N-acetylglucosaminidase, phosphatase and arylsulfatase) 활성도, 미생물 군집구조(eubacteria, denitrifying bacteria)의 특성을 알아보았다. 실험 결과 수변 및 사주 지표수-지하수 혼합대 토양이 많은 수분을 함유하고 있었고, 수분함량과 용존 유기탄소는 양의 상관관계를 보였으며, 체외 미생물 효소 활성도는 용존유기탄소와 양의 상관관계를 보였다. 진정세균 군집구조 또한 수변 및 사주의 지표수-지하수 혼합대에서 독특한 경향을 보였다. 이와 같은 결과는 지표수-지하수 혼합대를 통해 일어나는 하천의 생지화학적인 순환 전체를 이해하기 위한 기반을 제공한다.

• 생명과학회지
• /
• 제17권3호통권83호
• /
• pp.386-395
• /
• 2007

본 연구는 1999년에서 2001년도 걸쳐서 3년간 국내에서 분리된 환자유래 장염비브리오 18균주와 일본 후쿠오카 지역에서 2002년도에 환자에서 분리한 장염비브리오 9균주 등 총 27균주에 대하여 toxR 유전자의 검출, 혈청형별 검사, 약제내성의 양상, tdh, trh1 및 trh2 유전자의 보유상태 및 urease 생성성을 살펴보고, 혈청형 O3:K6 균주에 대하여 TDH의 생성성 검사, tdh 양성균주의 RFLP 형별, ORF 8의 분포, PFGE법과 RAPD법을 실시하였으며 결과는 다음과 같다. 1. 한국 및 일본의 환자유래 균주 대부분에서 urease음성이었으며, toxR 유전자로 확인 동정하였고 혈청형의 분포는 국내 분리 주의 O3:K6, O4:K9, O6:K46, O3:K57, O5:Kl5와 일본 분리주의 O3:K6, O1:K38, O4:K68, O4:Kl2의 혈청형으로 나타났다. 2. 항생제 감수성 시험에서는 vancomycin과 oxacillin은 27균주 (100%), penicillin은 26균주 (96.3%)로 높은 내성을 나타내었고, 14균주 (51.9%)가 4가지 이상의 항생제에 다제내성을 나타내었다. 항생제의 내성양상은 6종류로 혈청형에 관계없이 vancomycin, oxacillin, penicillin에 내성을 나타내는 V형이 15균주 (55.6%)로 가장 많이 나타났다. 3. tdh 유전자는 26균주가 PCR법과 DNA probe hybridization법의 결과에서 양성으로 나타났으며, urease양성이었던 일본 환자유래 혈청형 O3:K6 1균주만이 tdh유전자 음성, trh2 유전자 양성을 나타냈다. 혈청형 O3:K6의 TDH 생성성역가는 전 균주가 256배에서 2,048배 정도로 나타났으며, RFLP 양상은 모든 균주가 11.5 kb와 4.3 kb에 tdh 유전자를 보유하고 있었다. 또한 혈청형 O3:K6 균주들은 RAPD법과 PFGE법으로 유전자형별을 비교 검토한 결과 8형으로 거의동일한 유전자형별의 결과를 나타내었다. 4. ORF 8의 분포는 혈청형 O3:K6 전 균주에서 양성이었고, 특히 혈청형 O6:K46 4균주 모두에서 ORF 8 양성을 나타내어 새롭게 나타난 유행균주일 가능성을 시사 하였다. 따라서 ORF 8 유전자의 검출은 범세계적으로 유행하는 균주들을 동정하는데 있어 genetic marker로서 매우 유용한 것으로 사료된다.