• Title/Summary/Keyword: strain-specific condition

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Polymerase chain reaction for the detection of Newcastle disease virus (닭 뉴캐슬병 바이러스의 특이 검출을 위한 polymerase chain reaction 법)

  • Yeo, Sang-geon;Kim, Do-kyoung;Park, Seon-ja
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.565-573
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    • 1998
  • To study the specific tools for the diagnosis of Newcastle disease virus (NDV) in chicken, polymerase chain reaction (PCR) and its presumable conditions were evaluated for the detection of hemagglutinin-neuraminidase (HN) gene of NDV RNA. For these purposes, Kyojeongwon strain of the NDV was propagated in allantoic cavity of SPF embryonating chicken eggs, and viral RNA was extracted from fractionated virus after the allantoic fluids were ultracentrifuged with sucrose gradient. The first-strand cDNA was then made for the HN gene of NDV RNA by reverse transcription at $42^{\circ}C$ for 1 hour using specific primer complementary to the HN gene. The single-stranded cDNA was used as template in the PCR of the HN-DNA, and various conditions of the PCR were evaluated to set up method for the specific detection of the HN-DNA. The PCR conditions promising for the detection of HN gene consist of preheating at $94^{\circ}C$, 5 min, 30 cycles of denaturation at $94^{\circ}C$, 1 min, annealing at $55^{\circ}C$, 1 min and polymerization at $72^{\circ}C$, 2 min, and a cycle of extension at $72^{\circ}C$, 5 min. when NDVs of allantoic fluids without fractionation were applied to the above PCR condition, the HN genes were detected effectively not only from Kyojeongwon but from other velogenic strains such as Herts and a field isolate.

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Molecular Detection and Characterization of Orf Virus from Outbreak of Contagious Pustular Dermatitis in Korean Indigenous Goats (한국 재래 산양의 전염성 농피성 피부병에서 orf virus의 검출과 B2L 유전자를 통한 계통발생학적 분석)

  • Park, Jin-Ho;Kim, Guk-Jong;Choi, Wook;Kim, Eun-Ha;Han, Jae-Chul;Ou, Sung-Guk;Lee, Jon-Hwa;Cho, Mae-Rim;Song, Hee-Jong;Chae, Jun-Seok
    • Journal of Veterinary Clinics
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    • v.21 no.2
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    • pp.102-108
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    • 2004
  • Orf virus (ORFV), a member of genus Parapoxvirus (family-Poxviridae), a causative agent of contagious ecthyma in sheep and goat leading to a condition commonly known as vesicular dermatitis. Recently, twelve goats from Iksan in Jeonbuk province were observed with clinical signs like necrotic vesicular lesions around the mucosa of mouth, nasal cavity, eye, ear, teats, abdomen and groin. Based on these clinical symptoms, contagious ecthyma infection was suspected. The skin scrapping was collected from lesions for isolation of DNA and subsequent PCR amplification of ORFV specific 235 bp region of B2L gene. All of the samples were found positive by PCR analysis. Sequencing and further phylogenetic analysis of the PCR product revealed 100% identity to Japan isolate of ORFV (Okinawa, GenBank accession number AB080769), and showed 99.6% of similarity to New Zealand strain (NZ-2, GenBank accession number U06671). It was concluded that ORFV strain detected in the present study is homologous to Japan isolate and New Zealand strain. The PCR test based on amplification of B2L gene is a highly useful tools for rapid and specific diagnosis of contagious ecthyma.

Study of Viral Effects of the Mycovirus (LeV) and Virus-Free Commercial Line in the Edible Mushroom Lentinula edodes

  • Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.37-37
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    • 2015
  • dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

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Evaluation of spring shape effect on the nuclear fuel fretting using worn area (핵연료 프레팅 마멸에서 마멸면적을 이용한 스프링 형상 영향 평가)

  • Lee Young-Ho;Kim Hyung-Kyu;Jung Youn-Ho
    • Proceedings of the Korean Society of Tribologists and Lubrication Engineers Conference
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    • 2003.11a
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    • pp.313-323
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    • 2003
  • The sliding wear behaviors of Zircaloy-4 nuclear fuel rod were investigated using two support springs with convex and concave shapes in room temperature air and water. The main focus is to compare the wear behavior of various test variables such as slip amplitude, environment, contact contours with different spring shape and a number of cycles. The results indicated that wear volume and maximum wear depth increased with slip amplitude in both air and water, but their trends tended to change according to the spring shapes and test environments. In air condition, the wear volume was controlled by wear debris behavior generated on worn surface. As a result, final wear volume and maximum wear depth decreased if a ratio of protruded wear volume to worn area $(D_p)$ would be saturated to specific value. This is because wear particle layer could accommodate large strain by accumulating and transforming wear particle layer. However, in water condition, metal-to metal contact was more dominant and wear volume was greatly affected by changed mechanical behavior between contact surfaces since wear debris should be generated after repeated plastic deformation and fracture. After wear test, worn surfaces were examined using optical microscope and SEM and details of wear mechanism were discussed using a ratio of wear volume to worn area $(D_e)$ at each test condition.

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Studies on the Brewing of Apple Wine -Culture Conditions of a Cider Yeast, Saccharomyces sp. R-11 on the Synthetic Medium (사과주(酒) 양조(釀造)에 관한 연구(硏究) -사과주효모(酒酵母) Saccharomyces sp. R-11의 합성배지((合成培地)에서의 배양(培養) 조건(條件)-)

  • Chung, Ki-Taek;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.10 no.2
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    • pp.75-83
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    • 1982
  • As a primary study for cell growth and alcohol production of a cider yeast, Saccharomyces sp. R-11, cultural and nutritional characteristics of the strain were investigated. The results obtained were as follows: The optimum culture medium for this strain was a synthetic medium, Henneberg B, and sucrose was the best carbon source for yeast growth and alcohol production. Optimum sugar concentrations for yeast growth and alcohol production were 15% and 25%, respectively. Optimum pH and temperature of the basal medium for growth of this strain were 4.5 and $30^{\circ}C$ respectively. The yeast growth was enhanced by the addition of 100 ppm of $Mg^{2+}$, but significantly inhibited by the addition of 100 ppm of $Co^{2+}$. Lower temperature and maintenance of optimum pH for yeast growth increased the final alcohol concentration. Under optimum condition for cell growth at stationary culture, generation time and specific growth rate of the strain were 7.5 hr and 0.092 $hr^{-1}$, respectively. At 8% initial alcohol concentration, yeast growth was inhibited about 50% and this strain could not be grown at more than 12% initial alcohol. The strain could be grown at less than 125ppm $SO_2$without alcohol addition, and at less than 75 ppm $SO_2$ with 8% initial alcohol. The higher sulfur dioxide concentration of a medium, the longer lag phase in yeast growth was observed. This strain could induced alcoholic fermentation at less than 10% initial alcohol concentration with 0 and 25 ppm $SO_2$, at less than 8% initial alcohol with 50 and 75 ppm $SO_2$, and at less than 6% initial alcohol with 100 and 125 ppm $SO_2$.

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Purification of Alkaline Restriction Endonuclease from Alkalophilic Bacillus sp. 8-13 (제한효소 생성능을 지닌 알칼리성 Bacillus sp. 8-13 균주로부터 알칼리성 제한효소의 정제)

  • Bae, Moo;Lee, Jee-Eun;Park, Kyoung-Sook;Lee, Kang-Man
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.289-294
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    • 1992
  • Twenty-four bacterial strains among alkalophillic bacteria isolated from soil samples were examined for the presence of type II restriction endonuclease in aerobic culture. One strain was found to contain specific enzyme to cleave lambda DNA. The characteristics of this microorganism is the ability to grow well in alkalophilic and high temperature condition, that is at pH 10.3 and $50^{\circ}C$. This strain was tentatively identified to Bacillus alkaloPhilus subsp. halodurans when morphological, physiological and biochemical characteristics were examined. The enzyme was purified from crude extract by streptomycin sulfate, ammonium sulfate precipitation, which was followed by DEAE-cellulose and phosphocellulose ion exchange column chromatography, and the subunit molecular weight was about, 32,000 daltons by polyacrylamide gel electrophoresis containing 0.1% SDS.

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Sliding Wear Mechanism of Ultra-Fine Grained Low Carbon Dual Phase Steel as n Function of Applied Load (결정립 미세화에 따른 이상조직 탄소강의 하중에 따른 마멸 기구)

  • Yu, H.S.;Yi, S.K.;Shin, D.H.;Kim, Y.S.
    • Proceedings of the Korean Society for Technology of Plasticity Conference
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    • 2007.05a
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    • pp.421-424
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    • 2007
  • Dry sliding wear behavior of ultra-fine grained (UFG) plain low carbon dual phase steel, of which microstructure consists of hard martensite in a ductile ferrite matrix, has been investigated. The wear characteristics of the UFG dual phase steel was compared with that of a coarse grained dual phase steel under various applied load conditions. Dry sliding wear test were carried out using a pin-on-disk type tester at various loads of 1N to 100N under a constant sliding speed condition of 0.20m/s against an AISI 52100 bearing steel ball at room temperature. The sliding distance was fixed as 1000m for all wear tests. The wear rate was calculated by dividing the weight loss, measured to the accuracy of 10-5g by the specific gravity and sliding distance. The worn surfaces and wear debris were analyzed by SEM, EDS and profilometer. Micro-vickers hardness of the cross section of worn surfaces were conducted to analyze strain hardening underneath the contact surfaces. The wear mechanism of the UFG dual phase steel was investigated with emphasis on the unstable nature of the grain boundaries of the UFG microstructure.

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Study on the sexing of preimplantation mouse embryo exposed to H-Y ntisera I. Sexing of mouse embryos by cytolytic assay (H-Y항체에 의한 생쥐초기배의 성판별에 관한 연구 I. 세포발육능검사에 의한 성판별)

  • 양부근;김정익
    • Korean Journal of Animal Reproduction
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    • v.12 no.1
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    • pp.31-36
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    • 1988
  • There studies were conducted using inbred ICR mice to examine the sex of preimplantation mouse embryo. The morphological normality of mice embryos treated with the culture medium containing rat H-Y antiserum(10%, v/v) plus complement(20%,v/v) was observed and also the sexing of embryos was investigated by chromosomal analysis. The results obtained were summarized as follows: 1. The viability of preimplantation mouse embryos, which were incubated in vitro with different media condition, was scored 68.9-85.5% in control group. However, 151 embryos normally developed up to blastocyst and 160 embryos were retarded growth or destroyed out of total 311 embryos treated in the medium containing H-Y antiserum(10%, v/v) plus complement(20%,v/v). 2. H-Y antiserum was prepared from inb red rats (Wistar and Donryu strain) with different immunization times (4, 5 and 6th) to examine the specific titer of embryos by the number of immunization. Precentage of normally developed embryos incubated either in the medium containing the antiserum of Wistar plus complement or Donryu plus complement was revealed 50.9, 47.4 and 50.0% (4, 5 and 6th immunization and 47.8, 41.2 and 48.7%, respectively. 3. Twenty two females and five males were identified out of fourty-eight normally developed embryos incubated in the medium containing H-Y antiserum plus complement by chromosomal analysis.

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ACOUSTIC EMISSION CHARACTERISTICS OF STRESS CORROSION CRACKS IN A TYPE 304 STAINLESS STEEL TUBE

  • HWANG, WOONGGI;BAE, SEUNGGI;KIM, JAESEONG;KANG, SUNGSIK;KWAG, NOGWON;LEE, BOYOUNG
    • Nuclear Engineering and Technology
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    • v.47 no.4
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    • pp.454-460
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    • 2015
  • Acoustic emission (AE) is one of the promising methods for detecting the formation of stress corrosion cracks (SCCs) in laboratory tests. This method has the advantage of online inspection. Some studies have been conducted to investigate the characteristics of AE parameters during SCC propagation. However, it is difficult to classify the distinct features of SCC behavior. Because the previous studies were performed on slow strain rate test or compact tension specimens, it is difficult to make certain correlations between AE signals and actual SCC behavior in real tube-type specimens. In this study, the specimen was a AISI 304 stainless steel tube widely applied in the nuclear industry, and an accelerated test was conducted at high temperature and pressure with a corrosive environmental condition. The study result indicated that intense AE signals were mainly detected in the elastic deformation region, and a good correlation was observed between AE activity and crack growth. By contrast, the behavior of accumulated counts was divided into four regions. According to the waveform analysis, a specific waveform pattern was observed during SCC development. It is suggested that AE can be used to detect and monitor SCC initiation and propagation in actual tubes.

Sliding Wear Mechanism of Ultra-Fine Grained Low Carbon Dual Phase Steel as a Function of Applied Load (경정립 미세화에 따른 이상조직 탄소강의 하중에 따른 마멸기구)

  • Yu, H.S.;Yi, S.K.;Shin, D.H.;Kim, Y.S.
    • Transactions of Materials Processing
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    • v.16 no.4 s.94
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    • pp.299-303
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    • 2007
  • Dry sliding wear behavior of ultra-fine grained(UFG) plain low carbon dual phase steel, of which microstructure consists of hard martensite in a ductile ferrite matrix, has been investigated. The wear characteristics of the UFG dual phase steel was compared with that of a coarse grained dual phase steel under various applied load conditions. Dry sliding wear test were carried out using a pin-on-disk type tester at various loads of 1N to 100N under a constant sliding speed condition of 0.20m/s against an AISI 52100 bearing steel ball at room temperature. The sliding distance was fixed as 1000m for all wear tests. The wear rate was calculated by dividing the weight loss, measured to the accuracy of 10-5g by the specific gravity and sliding distance. The worn surfaces and wear debris were analyzed by SEM, EDS and profilometer. Micro-vickers hardness of the cross section of worn surfaces were conducted to analyze strain hardening underneath the contact surfaces. The wear mechanism of the UFG dual phase steel was investigated with emphasis on the unstable nature of the grain boundaries of the UFG microstructure.