• Title/Summary/Keyword: PCR 동정

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Identification of Bacteria by Sequence Analysis of 16S rRNA in Testes of Jeju Horses (제주마 고환내 세균의 16S rRNA 염기서열 분석을 이용한 동정)

  • Park, Yong-Sang;Kim, Nam-Young;Han, Sang-Hyun;Park, Nam-Geon;Ko, Moon-Suck;Cho, Won-Mo;Chae, Hyun-Seok;Cho, In-Chul;Cho, Sang-Rae;Woo, Jae-Hoon;Kang, Tae-Young
    • Journal of Veterinary Clinics
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    • v.31 no.1
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    • pp.36-39
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    • 2014
  • Many bacteria colonized in the horse semen affect quality of the sperm and some may cause infection in the mare reproductive tract and infertility of susceptible mare. This study was initiated to determine the prevalence of bacteria in testes of Jeju horses by determining rRNA sequence. The samples were swabed from the testes of nine Jeju horses (aged from 8 to 12 months after birth). Bacteria isolated from testes were identified by 16S rDNA sequencing. 1.6-kbp PCR products for 16S rRNA coding region were obtained using the universal primers. The PCR products were further purified and sequenced. Maximum similar species were found by BLAST search in the GenBank DNA database. BLAST results showed that the sequences were similar to those of Acinetobacter sp (A. schindleri, A. ursingii)., Bacillus cereus, Corynebacterium glutamicum, Escherichia coli, Gamma proteobacterium, Micrococcus luteus, Pseudomonas mendocina, Shigella sonnei, Sphingomonas sp., Staphylococcus sp (S. cohnii, S. saprophyticus, S. xylosus)., and Stenotrophomonas maltophilia. DNA sequences for 16S rRNA is provided useful informations for species identification of pathogenic microorganisms for the reproductive organs in horses.

Outbreak Pattern of Isolated Enterovirus Causing Aseptic Meningitis in Busan, 2002 Years. (2002년도 바이러스성 뇌수막염의 발생양상)

  • 조경순;정명주
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.179-183
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    • 2004
  • Enterovirus isolation was attempted from samples obtained from aseptic meningitis-suspected patients in hospitals in Busan during 2002. Enteroviruses were found in 83 of 703 cases. Echovirus serotypes 2, 3, 6, 7, 9, 13, 25, 29, 30 were isolated in 72 cases while coxsackievirus serotypes B3 and B4 were isolated in 11 cases. The occurrence was found to be distributed from April thru November with the highest rate during June and July. The strains of Vero and HEp-2 of echovirus and coxsackievirus, respectively, are highly infectious. Age distribution of patients, 61 patients in Enteroviruses and 11 patients in coxsackievirus, respectively, occurred in 0 to 10 year-olds. The sex distribution of the patients is as follows, 52 males (62.7%), 31 females (37.3%). Occurrence rate was found to be higher in male patients. Electron micrograph of echovirus and coxsackievirus show that they are small nonenevolped, isometric-shaped viruses. Isolated RNA from strains of echovirus and coxsackievirus showing cytopathic effects were used to undergo nested PCR which resulted in a 437 bp single band in all the strains.

DETECTION OF BLACK-PIGMENTED BACTERIA IN INFECTED ROOT CANALS (감염근관에서 Black-Pigmented Bacteria의 동정에 대한 연구)

  • Kwon, Eun-Kyoung;Kim, Eun-Sook;Kwak, Ju-Seog;Lee, Hwang;Lee, Su-Jong;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.27 no.1
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    • pp.54-65
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    • 2002
  • Black-pigmented bacteria have been implicated in the endodontic infections. This group of microorganisms includes Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. The organisms display a wide variety of virulence factors that may be pertinent to acute endodontic infections. The aim of this study was to identify P. endodontalis, P. gingivalis, P. intermedia, and P. nigrescens by using special potency disk test, filter paper spot test, 165 rRNA gene-directed PCR, and API 32A. Microbial samples were collected from root canals of 33 intact teeth with necrotic pulp and/or apical periodontitis. Conventional laboratory methods were used for identification of the strains of black pigmented bacteria. Eighteen of 33 samples were positive for the growth of black-pigmented bacteria Five colonies were cultured from each pure cultured colonies from Brucella agar plate. Seventy seven colonies were positive for the growth of black-pigmented bacteria. Thirty three of 77(42.6%) were identifed as P. nigrescens, 10 of 77(12.9%)were P. gingivalis, 6 of 77(7.8%) were P. endodontalis, 10 of 77(12.9%) were P. intermedia. On the contrary the reference strains of P. nigrescens, experimental strains of P nigrescens was sensitive to kanamycin in special potency disk test. 165 rRNA gene PCR and API test after rapid presumptative identification methods, such as special potency disk test and filter paper spot test, would be accurate detection methods for black-pigemented bacteria.

Characterization of Phosphatidylinositol Glycan, Class K (PIGK) Gene and Analysis of Association with Quantitative Traits in Pigs (돼지 Phosphatidylinositol Glycan, Class K (PIGK) 유전자의 동정과 양적형질과의 연관성 분석)

  • Lim, H.T.;Kim, J.H.;Choi, B.H.;Lee, S.H.;Park, E.W.;Kim, T.H.;Cho, I.C.;Oh, S.J.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.47 no.2
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    • pp.167-176
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    • 2005
  • PIGK(phosphatidylinositol glycan, class K) is a subunit of GPI transamidase that cleaves the signal peptide in proproteins and replaces it with GPI. In addition, the structure and synthesis of GPI are critically involved in some of the cellular actions of insulin. Therefore, PIGK would be essential for mammalian development and many specific cellular functions as well as for metabolic activity of insulin associated with GPI. Two types of" full-length cDNAs of porcine PIGK were cloned through RT-PCR and RACE experiments. One is thought to be a normal form(consist of 395 amino acids) and the other is considered as an alternative spliced form(consist of 371 amino acids) which contains additional 63 bps in intron 7. Since a stop codon was contained within the insertion, the spliced form has a shorter coding sequence than that of normal form. A missense mutation (T314I) in exon 6 was detected and used for genotyping to estimate association with the growth and fat deposition traits for 545 $F_2$ animals(Korean native boars ${\times}$ Landrace). From the PCR-RFLP analysis using HpyCH4III, CT genotype showed highly significant relationship(P< 0.01) with carcass fat contents.

Biovars of Pseudomonas syringae pv. actinidiae Strains, the Causal Agent of Bacterial Canker of Kiwifruit, Isolated in Korea (우리나라에서 분리한 참다래 궤양병균 Pseudomonas syringae pv. actinidiae 균주들의 Biovar)

  • Lee, Young Sun;Kim, Jin;Kim, Gyoung Hee;Choi, Eu Ddeum;Koh, Young Jin;Jung, Jae Sung
    • Research in Plant Disease
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    • v.23 no.1
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    • pp.35-41
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    • 2017
  • Pseudomonas syringae pv. actinidiae is the causative agent of bacterial canker of kiwifruit. The population of this pathogen is differentiated into three biovars, biovar 1, 2 and 3, according to their molecular characteristics. In this work, we determined biovars of P. syringae pv. actinidiae strains isolated in Korea since 1997 and stored in Department of Biology, Sunchon National University, Suncheon, Korea. The biovars of P. syringae pv. actinidiae strains were determined by PCR using biovar specific primers developed previously. Of 682 strains investigated, 288 strains belonged to biovar 2, while 394 strains were biovar 3. There were no P. syringae pv. actinidiae strains belonging to biovar 1 among the strains isolated in Korea. Sudden outbreak and spreading of bacterial canker caused by biovar 3 strain suggest that this strain has character of rapid transmission.

Isolation, identification and serological investigation of Actinobacillus pleuropneumoniae in slaughtered pigs (도축돈에서의 Actinobacillus pleuropneumoniae 분리, 동정 및 감염률 조사)

  • Kim, Kyung-Eon;Ku, Kyung-Nyer;Ko, Jae-Hyung;Moon, Hyeong-Jun;Choi, Kwon-Rag;Song, Eun-Ah;Park, Mi-Young
    • Korean Journal of Veterinary Service
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    • v.36 no.3
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    • pp.181-186
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    • 2013
  • This study was conducted to isolate the Actinobacillus pleuropneumoniae (APP) and to find out the distribution of 15 serovars mainly in southern Gyeonggi province, Korea. From July 2011 to Nov. 2012, a total of 2,204 slaughter pigs (110 herds) were inspected for evaluation of APP like pneumonic lesions. 48 (33.8%) APP strains were isolated from the 142 lungs and identified using PCR assays (cps, apx/omlA, biovar). Consequently, the serotype ratio were as in the following; type2 41.7% (n=20), type5 33.3% (n=16), type12 10.4% (n=5), type1 6.2% (n=3), type4 and 7 2.1% (n=1) and unknown 4.2% (n=2). Also serological test was implemented for 452 (83 herds) serum samples randomly collected from above slaughter pigs using commercial ELISA kits. The positive ratio of each serotype for tested pigs were 19.1% (77/404) on [2], 7.1% (32/452) on [3, 6, 8], 6.9% (28/404) on [5a, 5b], 6.2% (28/452) on [4, 7], 2.8% (9/320) on [12], 2.0% (9/452) on [1, 9, 11] and 0.0% (0/452) on [10]. And 49.3% (223/452) of pigs were positive on apxIV antibody. On the basis of latter screening test, the infected farm ratio accounted for 71.1% (59/83) and that was much higher than previously reported data.

Unrecorded fungi isolated from Lentinula edodes cultivation houses in Korea (표고 재배사에서 분리한 국내 미기록 진균 보고)

  • Ahn, Geum Ran;Noh, Hyeungjin;Kim, Jun Young;Ko, Han Gyu;Kim, Seong Hwan
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.72-78
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    • 2020
  • During the monitoring of fungal pests in 2016 and 2017, Acrodontium crateriforme, Naganishia friedmannii, Pestalotiopsis trachicarpicola, Penicillium wollemiicola, and Trichoderma thailandicum were isolated from indoor air, mushroom flies (Phytosciara flavipes), and media materials in the cultivation houses of oak wood mushroom (Lentinula edodes) located in Seocheon, Jangheung, Buyeo, and Yeoju, Korea. These fungal species were identified based on their morphological characteristics after their growth on PDA and subsequent molecular analyses of the 26S rDNA, 28S rDNA, β-tubulin gene, and translation elongation factor 1-α gene using PCR amplification and nucleotide sequencing were performed. The results showed that these fungi were previously undocumented in Korea. This study reports descriptions of their taxonomical and known properties.

Phytophthora citricola, a Causal Agent of Jujube (Zizyphus jujuba) Fruit Rot (대추 역병균의 Phytophthora citricola의 동정)

  • 지형진;임양숙;정기채;조원대
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.402-407
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    • 1998
  • Phytophthora rot on jujube fruit has occurred at several cultivation areas in Kyung-buk and Kyung-nam provinces. Symptoms consisted of brownish to reddish rot on fruits resulting in early drop or mummification. The causal fungus isolated from infected fruits and adjacent leaf stalks was identified as Phytophthora citricola, which has never been reported in Korea. Sporangia were semi-papillate, noncaducous and highly variable in shapes. Plerotic oospores with paragynous antheridia were abundant is single cultures. Sporangia of two isolates were measured as 38-76$\times$20-40 ${\mu}{\textrm}{m}$ and averaged 51.4$\times$27.0 and 55.6$\times$36.0 ${\mu}{\textrm}{m}$. Oogonia were ranged from 26 to 36 ${\mu}{\textrm}{m}$ and averaged 31.3 and 32.0 ${\mu}{\textrm}{m}$. Colony pattern was slightly radiated with sparse aerial mycelia on common media. Minium, optimum and maximum temperatures for mycelial growth were recorded at 7, 25, and 32$^{\circ}C$, respectively. Among tested media, 10% V8A was the best and $25^{\circ}C$ was better than 15$^{\circ}C$ for oospore formation of the fungus. The jujube isolates of P. citricola were readily differentiated from other closely related species in the genus, namely; P. nicotianae, P. citrophthora, P. cactorum, P. capsici, and P. plalmivora on the basis of PCR-RFLP of r-DNA. The fungus showed strong pathogenicty to jujube, apple, pear, orange, persimmon and eggplant, and relatively weak to citron, tomato, pepper and cucumber. In this study, P. citrocola is firstly identified and jujube fruit rot caused by the fungus is recorded as a new disease in Korea.

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Identification of the Black-pigmented Prevotella Species and Porphyromonas Species from Infected Dental Root Canals (치아 근관 감염 검체에서 검은 색소 형성 Prevotella species와 Porphyromonas species의 동정)

  • Kim, Eun-Sook;Kim, Shin-Moo
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.1
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    • pp.45-53
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    • 2006
  • Anaerobic black-pigmented bacteria have been implicated in the endodontic infections. This group of microorganisms includes Porphyromonas endodontalis, P. gingivalis, Prevotella intermedia, and Prevotella nigrescens. The organisms display a wide variety of virulence factors that may be pertinent to acute endodontic infections. The aim of this study was to identify P. endodontalis, P. gingivalis, P. intermedia, and P. nigrescens by using the special potency disk test, filter paper spot test, 16S rRNA gene-directed PCR, and API 32A system. Microbial samples were collected from root canals of 33 intact teeth with necrotic pulp and apical periodontitis. Conventional laboratory methods were used to identify the strains of anaerobic black pigmented bacteria. Eighteen out of 33 samples were positive for the growth of black-pigmented bacrteria. Five colonies were cultured from each pure cultured colony from Brucella agar plates. Seventy seven colonies were positive for the growth of black-pigmented bacteria. Thirty three out of 77(42.8%) were identifed as P. nigrescens, 10 out of 77(13%)were P. gingivalis, 6 out of 77(7.8%) were P. endodontalis, 10 out of 77(13%) were P. intermedia. On the contrary the reference strains of P. nigrescens, experimental strains of P. nigrescens were susceptible to kanamycin in the special potency disk test. We concluded that after rapid presumptive identification methods, such as the special potency disk test and filter paper spot test were done, 16S rRNA gene PCR and API 32A test would be accurate detection methods for black-pigemented bacteria.

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A Study for the Expression of Melatonin Receptor Gene and Reproductive Indices in Golden Hamsters Exposed to Photoperiods (골든 햄스터에서 광주기에 따른 멜라토닌 수용체 유전자 발현과 생식 지수들에 관한 연구)

  • Choi, Donchan;Choi, Hyungjae;Lim, Sinae;Park, Changeun
    • Development and Reproduction
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    • v.6 no.1
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    • pp.45-54
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    • 2002
  • Reproductive activity of golden hamsters(Mesocricetus auratus) is regulated by the photoperiod. They are sexually active in summer and inactive in winter. Melatonin, a pineal hormone, has been known to mediate sexual activities in seasonal breeding animals. Melatonin receptor was recently identified in several animal species including hmm. But little has been known about it in relation to the reproductive activities of golden hamsters. By using reverse transcription polymerase chain reaction(RT-PCR) methods, a portion of the melatonin receptor gene(309 nucleotides) was identified in golden hamsters. The nucleotide sequence of the melatonin receptor and the amino acid sequence deduced were compared to those reported in other animals. Melatonin receptors were obviously detected in hypothalamus, pituitary containing pars tuberalis, blood, and spleen. Although the testicular weights and the levels of reproductive hormones were dramatically affected by photoperiods, the expression of melatonin receptor was not markedly changed by them. These results suggest that the action of melatonin in regulating reproduction might be mainly due to the affinity of melatonin receptor rather than the density fi melatonin receptor.

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