• Title/Summary/Keyword: Strain Amplification

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Isolation and Characterization of 4-(2,4-Dichlorophenoxy)Butyric Acid-Degrading Bacteria from Agricultural Soils

  • Park, In-Hyun;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.243-250
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    • 2003
  • Eight numerically dominant 4-(2,4-dichlorophenoxy) butyric acid (2,4-DB)-degrading bacteria and three pairs of bacteria showing syntrophic metabolism of 2,4-DB were isolated from soils, and their phylogenetic and phenotypic characteristics were investigated. The isolates were able to utilize 2,4-DB as a sole source of carbon and energy, and their 2.4-DB degradative enzymes were induced by the presence of 2.4-DB. Analysis of 16S rDNA sequences indicated that the isolates were related to members of the genera, Variovorax, Sphingomonas, Bradyrhizobium, and Pseudomonas. The chromosomal DNA patterns of the isolates obtained by polymerase-chain-reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences were distinct from each other. Four of the isolates had plasmids, but only one strain, DB 1, rad a transmissible 2,4-D degradative plasmid. When analyzed with PCR using primers targeted to the tfdA, B, and C genes, only strains DB2 and DB9a produced DNA bands of the expected sizes with the tfdA and C primers, respectively. All of the isolates were able to degrade 2,4-D as well as 2,4-DB, suggesting that the degradation pathways of these compounds were closely related to each other, but respiratory activities of many isolates adapted to 2,4-DB metabolism were quite low with 2,4-D.

Evaluation of Dorim-Goh bridge using ambient trucks through short-period structural health monitoring system

  • Kaloop, Mosbeh R.;Hwang, Won Sup;Elbeltagi, Emad;Beshr, Ashraf;Hu, Jong Wan
    • Structural Engineering and Mechanics
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    • v.69 no.3
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    • pp.347-359
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    • 2019
  • This paper aims to evaluate the behavior of Dorim-Goh bridge in Seoul, Korea, under static and dynamic loads effects by ambient trucks. The prestressed concrete (PSC) girders and reinforcement concrete (RC) slab of the bridge are evaluated and assessed. A short period monitoring system is designed which comprises displacement, strain and accelerometer sensors to measure the bridge performance under static and dynamic trucks loads. The statistical analysis is used to assess the static behavior of the bridge and the wavelet analysis and probabilistic using Weibull distribution are used to evaluate the frequency and reliability of the dynamic behavior of the bridge. The results show that the bridge is safe under static and dynamic loading cases. In the static evaluation, the measured neutral axis position of the girders is deviated within 5% from its theoretical position. The dynamic amplification factor of the bridge girder and slab are lower than the design value of that factor. The Weibull shape parameters are decreased, it which means that the bridge performance decreases under dynamic loads effect. The bridge girder and slab's frequencies are higher than the design values and constant under different truck speeds.

Molecular Typing of Leuconostoc citreum Strains Isolated from Korean Fermented Foods Using a Random Amplified Polymorphic DNA Marker

  • Kaur, Jasmine;Lee, Sulhee;Sharma, Anshul;Park, Young-Seo
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.174-179
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    • 2017
  • For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Mornitoring and Identification of Human Astrovirus from Groundwater in Korea Based on Highly Sensitive RT-nested PCR Primer Sets

  • Lee, Siwon;Bae, Kyung Seon;Park, Jihyun;Kim, Jin-Ho;Lee, Jin-Young;Choi, Jiwon;Park, Eung-Roh;You, Kyung-A
    • Biomedical Science Letters
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    • v.27 no.4
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    • pp.255-263
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    • 2021
  • Human Astrovirus (HuAstV) is an important gastrointestinal pathogen that is frequently reported worldwide. Monitoring of contaminated groundwater has been suggested since HuAstV is transmitted through the fecal-oral route. This study developed a test method based on conventional reverse transcription (RT)-nested polymerase chain reaction (PCR) that involves SL® non-specific reaction inhibitor for unknown non-specific amplification taking place in the groundwater environment. An optimal method for detecting HuAstV in groundwater sample through analysis and comparison against conventionally reported method was also suggested. The developed method enabled the production of nested PCR amplicon of 630 nt, which is a sufficient length for similarity analysis based on sequencing and genotyping. Amplicons suspected to be HuAstV were amplified in two out of the twenty groundwater samples collected in Korea, presenting 99.77% and 99.73% similarity against HuAstV 1 strain lhar/2011/kor (JN887820.1) in sequencing, respectively. These amplicons were identified as HuAstV 1.

Finite element analysis of ratcheting on beam under bending-bending loading conditions

  • Sk. Tahmid Muhatashin Fuyad;Md Abdullah Al Bari;Md. Makfidunnabi;H.M. Zulqar Nain;Mehmet Emin Ozdemir;Murat Yaylaci
    • Structural Engineering and Mechanics
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    • v.89 no.1
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    • pp.23-31
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    • 2024
  • Ratcheting is the cyclic buildup of inelastic strain on a structure resulting from a combination of primary and secondary cyclic stress. It can lead to excessive plastic deformation, incremental collapse, or fatigue. Ratcheting has been numerically investigated on a cantilever beam, considering the current study's primary and secondary bending loads. In addition, the effect of input frequency on the onset of ratcheting has been investigated. The non-linear dynamic elastic-plastic approach has been utilized. Analogous to Yamashita's bending-bending ratchet diagram, a non-dimensional ratchet diagram with a frequency effect is proposed. The result presents that the secondary stress values fall sequentially with the increase of primary stress values. Moreover, a displacement amplification factor graph is also established to explain the effect of frequency on ratchet occurrence conditions. In terms of frequency effect, it has been observed that the lower frequency (0.25 times the natural frequency) was more detrimental for ratchet occurrence conditions than the higher frequency (2 times the natural frequency) due to the effect of dynamic displacement. Finally, the effect of material modeling of ratcheting behavior on a beam is shown using different hardening coefficients of kinematic hardening material modeling.

Mband characterization of a cultivar "DanBi 5Ho" with a long shelf life (큰느타리버섯 단핵균주 간 교잡에 의한 저장우수형 품종 "단비5호" 육성 및 특성)

  • Lee, Young-Han;Kim, Ya-El;Seuk, Su-Won;Jeong, Jeong-Min;Ryu, Jae-San;Heo, Jae-Young;Kim, Hee-Dae;Choe, Yong-Jo;Kim, Min-Keun
    • Journal of Mushroom
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    • v.14 no.2
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    • pp.64-69
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    • 2016
  • Pleurotus eryngii is one of the most widely cultivated and important edible mushrooms. However, the shelf life of the fruiting body is short, which. To solve this problem, a new cultivar, DanBi 5Ho, of Pleurotus eryngii was developed by mono-mono crossing between monokaryotic strains derived from KNR2598 and KNR2610. The optimum temperature formycelial growth was 25 and that forfruiting body development was 15-16. The quality did not change after a period of 40.0 d at 4. This result was significantly different compared to that ofthe control strain Knneuari 2 Ho. Analysis of genetic characteristics of the new hybrid strain DanBi 5Ho revealeda different profile thanthat of the parental and control strains when random amplification of polymorphic DNA (RAPD) primers we used. The results obtained from this study show that DanBi 5Ho is a new hybrid strain, characterized by improved storability after harvesting.

Cultural Conditions for Mycelial Growth and Molecular Phylogenetic Relationship in Different Wild Strains of Schizophyllum commune

  • Alam, Nuhu;Cha, Youn-Jeong;Shim, Mi-Ja;Lee, Tae-Soo;Lee, U-Youn
    • Mycobiology
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    • v.38 no.1
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    • pp.17-25
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    • 2010
  • The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

In Vivo Analysis of fadB Homologous Enzymes Involved in Biosynthesis of Polyhydroxyalkanoates in Recombinant Escherichia coli (재조합 대장균에서 fadB 유사효소의 Polyhydroxyalkanoates 합성에 미치는 역할의 규명)

  • 최종일;박시재;이상엽
    • KSBB Journal
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    • v.19 no.4
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    • pp.331-334
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    • 2004
  • In vivo characterization of FadB homologous enzymes including PaaG, YdbU and YgfG for medium-chain-length (MCL) polyhydroxyalkanoate (PHA) biosynthesis was carried out in fadB mutant Escherichia coli. Previously, it was reported that amplification of FadB homologous enzymes such as PaaG and YdbU in fadB mutant E. coli resulted in enhanced biosynthesis of MCL-PHA by greater than two fold compared with control strain. In this study, we constructed paaG fadB double mutant E. coli WB114 and ydbU fadB double mutant E. coli WB115 to investigate the roles of PaaG and YdbU in biosynthesis of MCL-PHA. Inactivation of paaG and ydbU genes in fadB mutant E. coli harboring Pseudomonas sp. 61-3 phaC2 gene reduced the MCL-PHA production to 0.16 and 0.16 PHA g/L, respectively from 2 g/L of sodium decanoate, which are much lower than 0.43 PHA g/L obtained with fadB mutant E. coli WB101 harboring the phaC2 gene. Also, we identified new FadB homologous enzyme YgfG, and examined its roles by overexpression of ygfG and construction of ygfG fadB double mutant E. coli WB113.

Gene Mutations of 23S rRNA Associated with Clarithromycin Resistance in Helicobacter pylori Strains Isolated from Korean Patients

  • Kim, Jung-Mogg;Kim, Joo-Sung;Kim, Na-Young;Kim, Yeoung-Jeon;Kim, In-Young;Chee, Young-Joon;Lee, Chul-Hoon;Jung, Hyun-Chae
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1584-1589
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    • 2008
  • Although resistance of Helicobacter pylori to clarithromycin is a major cause of failure of eradication therapies, little information is available regarding gene mutations of clarithromycin-resistant primary and secondary H. pylori isolates in Korea. In the present study, we examined gene mutations of H. pylori 238 rRNA responsible for resistance to clarithromycin. DNA sequences of the 238 rRNA gene in 21 primary clarithromycin-resistant and 64 secondary clarithromycin-resistant strains were determined by PCR amplification and nucleotide sequence analyses. Two mutations of the 238 rRNA gene, A2143G and T2182C, were observed in primary clarithromycin-resistant isolates. In secondary isolates, dual mutation of A2143G+T2182C was frequently observed. In addition, A2143G+T2182C+ T2190C, A2143G+T2182C+C2195T, and A2143G+T2182C+A2223G were observed in secondary isolates. Furthermore, macrolide binding was tested on purified ribosomes isolated from T2182C or A2143C mutant strains with $[^{14}C]$erythromycin. Erythromycin binding increased in a dose-dependent manner for the susceptible strain but not for the mutant strains. These results indicate that secondary isolates show a greater variety of 238 rRNA gene mutation types than primary isolates, and triple mutations of secondary isolates are associated with A2143G+T2182C in H. pylori isolated from Korean patients.

Taxonomic Characterization, Evaluation of Toxigenicity, and Saccharification Capability of Aspergillus Section Flavi Isolates from Korean Traditional Wheat-Based Fermentation Starter Nuruk

  • Bal, Jyotiranjan;Yun, Suk-Hyun;Chun, Jeesun;Kim, Beom-Tae;Kim, Dae-Hyuk
    • Mycobiology
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    • v.44 no.3
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    • pp.155-161
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    • 2016
  • The most economically important species used in a wide range of fermentation industries throughout Asia belong to Aspergillus section Flavi, which are morphologically and phylogenetically indistinguishable, with a few being toxigenic and therefore a major concern. They are frequently isolated from Korean fermentation starters, such as nuruk and meju. The growing popularity of traditional Korean alcoholic beverages has led to a demand for their quality enhancement, therefore requiring selection of efficient non-toxigenic strains to assist effective fermentation. This study was performed to classify the most efficient strains of Aspergillus section Flavi isolated from various types of traditional wheat nuruk, based on a polyphasic approach involving molecular and biochemical evaluation. A total of 69 strains were isolated based on colony morphology and identified as Aspergillus oryzae/flavus based on internal transcribed spacer and calmodulin gene sequencing. Interestingly, none were toxigenic based on PCR amplification of intergenic regions of the aflatoxin cluster genes norB-cypA and the absence of aflatoxin in the culture supernatants by thin-layer chromatography analysis. Saccharification capability of the isolates, assessed through ${\alpha}-amylase$ and glucoamylase activities, revealed that two isolates, TNA24 and TNA15, showed the highest levels of activity. Although the degrees of variation in ${\alpha}-amylase$ and glucoamylase activities among the isolates were higher, there were only slight differences in acid protease activity among the isolates with two, TNA28 and TNA36, showing the highest activities. Furthermore, statistical analyses showed that ${\alpha}-amylase$ activity was positively correlated with glucoamylase activity (p < 0.001), and therefore screening for either was sufficient to predict the saccharifying capacity of the Aspergillus strain.