• Journal of Microbiology
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• v.44 no.1
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• pp.42-49
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• 2006

In this study, we describe our newly-developed sensitive two-stage PCR procedure for the detection of 13 common mycoplasmal contaminants (M. arthritidis, M. bovis, M. fermentans, M. genitalium, M. hominis, M. hyorhinis, M. neurolyticum, M. orale, M. pirum, M. pneumoniae, M. pulmonis, M. salivarium, U. urealyticum). For primary amplification, the DNA regions encompassing the 16S and 23S rRNA genes of 13 species were targeted using general mycoplasma primers. The primary PCR products were then subjected to secondary nested PCR, using two different primer pair sets, designed via the multiple alignment of nucleotide sequences obtained from the 13 mycoplasmal species. The nested PCR, which generated DNA fragments of 165-353 bp, was found to be able to detect 1-2 copies of the target DNA, and evidenced no cross-reactivity with the generated DNA of related microorganisms or of human cell lines, thereby confirming the sensitivity and specificity of the primers used. The identification of contaminated species was' achieved via the performance of restriction fragment length polymorphism (RFLP) coupled with Sau3AI digestion. The results obtained in this study furnish evidence suggesting that the employed assay system constitutes an effective tool for the disagnosis of mycoplasmal contamination in cell culture systems.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.65-70
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• 2002

A TGF-${\beta}1$/GFP monomeric fusion protein was cloned from pPK9A and pGFP-Cl plasmid by PCR amplification. The fusion protein was expressed in a $Bac-To-Bac^{TM}$ baculovirus expression system. A 45 kDa fusion protein was purified using an Ni-NTA column with 300 mM imidazol from a cell lysate infected with recombinant viruses for 72 h post-infection. The fusion protein cross-reacted with the commercial $TGF-{\beta}1$ polyclonal Ab as well as Ab raised against a precursor, monomeric $TGF-{\beta}1$, and GFP. The binding activity of the fusion protein with a $TGF-{\beta}1$ receptor was examined. Fluorescence was observed in Mv1Lu cells, yet not in insect cells treated with the fusion protein. No fluorescence was detected in Mv1Lu cells incubated with the fusion protein treated with Ab prior to the binding reaction, or with GFP alone, thereby indicating that the binding of the fusion protein was specific to $TGF-{\beta}1$ with a receptor.

• Korea-Australia Rheology Journal
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• v.14 no.2
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• pp.57-62
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• 2002

The sensitivity of the final filament to the ongoing sinusoidal disturbances has been Investigated in the viscoelastic spinning using frequency response method. Amplification ratios or gains of the spinline cross-sectional area at the take-up to any disturbances show resonant peaks along the frequency regime, where the frequencies at theme points directly correspond to the imaginary parts of the successive leading eigenvalues from the linear stability analysis. As shown in Jung et al. (1999) and Lee et al (2001), the sensitivity results on the effect of various process conditions such as spinline cooling and fluid viscoelasticity, obtained by dynamic transient simulation have been corroborated in this study. That is, increasing spinline cooling makes the system less sensitive to disturbances, thus stabilizes the spinning. Also, an increasing viscoelasticity for extension-thickening fluids decreases the sensitivity of the spinning. i.e., stabilizing the system, where, as it increases the sensitivity of the spinning of extension-thinning fluids. Furthermore, it has been found in the present study that the inertia force as one of secondary forces causes the system to be more stabile or less sensitive to process disturbances.

• Korean Journal of Veterinary Research
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• v.57 no.1
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• pp.37-42
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• 2017

Getah virus (GETV) infection causes sporadic outbreaks of mild febrile illness in horses and reproductive failure in pigs. In this study, we established a reverse transcription polymerase chain reaction (RT-PCR) method to detect GETV from suspected virus-infected samples. The reaction conditions were optimized and validated by using RNA extracted from GETV propagated in cell culture. A GETV-specific GED4 primer set was designed and used to amplify a 177 bp DNA fragment from a highly conserved region of the E1 glycoprotein gene in the GETV genome. RT-PCR performed with this primer set revealed high sensitivity and specificity. In the sensitivity test, the GED4 primer set detected GETV RNA at the level of $10^{2.0}\;TCID_{50}/mL$. In the specificity test, the GED4 primer set amplified only a single band of PCR product on the GETV RNA template, without non-specific amplification, and exhibited no cross-reactivity with other viral RNAs. These results suggest that this newly established RT-PCR method is useful for accurate identification of GETV infection in animals.

• The Transactions of The Korean Institute of Electrical Engineers
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• v.57 no.12
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• pp.2300-2307
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• 2008

This study receives the noise transmitted in a constant audio frequency range through a microphone array in which the noise(like grease in a pan) occurs on the power supply line due to the troublesome partial discharge(arc). Then by going through a series of signal processing of removing noise, this study measures the distance and direction up to the noise caused by the troublesome partial discharge(arc) and monitors the result by displaying in the analog and digital method. After these, it determines the state of each size and judges the distance and direction of problematic part. When the signal sound transmitted by the signal source of bad insulator is received on each microphone, the signal comes only in the frequency range of 20 kHz by passing through the circuit of amplification and 6th low pass filter. Then, this signal is entered in a digital value of digital signal processing(TMS320F2812) through the 16-bit A/D conversion. By doing so, the sound distance, direction and coordinate of bad insulator can be detected by realizing the correlation method of detecting the arriving time difference occurring on each microphone and the algorithm of detecting maximum time difference.

• BMB Reports
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• v.41 no.11
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• pp.771-777
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• 2008

Calmodulin (CaM) proteins, members of the EF-hand family of $Ca^{2+}$-binding proteins, represent important relays in plant calcium signals. Here, OsCam1-1 was isolated by PCR amplification from the rice genome. The gene contains an ORF of 450 base pairs with a single intron at the same position found in other plant Cam genes. A promoter region with a TATA box at position-26 was predicted and fused to a gus reporter gene, and this construct was used to produce transgenic rice by Agrobacterium-mediated transformation. GUS activity was observed in all organs examined and throughout tissues in cross-sections, but activity was strongest in the vascular bundles of leaves and the vascular cylinders of roots. To examine the properties of OsCaM1-1, the encoding cDNA was expressed in Escherichia coli. The electrophoretic mobility shift when incubated with $Ca^{2+}$ indicates that recombinant OsCaM1-1 is a functional $Ca^{2+}$-binding protein. In addition, OsCaM1-1 bound the CaMKII target peptide confirming its likely functionality as a calmodulin.

• Korean Journal of Plant Taxonomy
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• v.50 no.1
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• pp.22-26
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• 2020

Chrysosplenium aureobracteatum Y. I. Kim & Y. D. Kim (Saxifragaceae) is a recently described endemic species growing in the central part of the Korean peninsula. It requires constant monitoring for conservation due to its limited distributions. There is also a need for molecular markers for proper assessments of the genetic differentiation of C. aureobracteatum from species morphologically similar to it. In this study, we developed microsatellite markers that can be used to evaluate the genetic diversity of this species, representing fundamental data with which to conserve the natural populations of the species. A total of 17 expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed by the Illumina pair-end sequencing of the transcriptomes of C. aureobracteatum. These markers were successfully applied to populations of C. aureobracteatum and to its most closely related species, C. barbatum, revealing high polymorphism in both species. The EST-SSR markers developed in this study were proven to be useful not only to monitor the population genetic structure of C. aureobracteatum for conservation purposes but also to study the genetic delimitation of the species from species closely related to it.

• Korea-Australia Rheology Journal
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• v.15 no.2
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• pp.91-96
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• 2003

The sensitivity of the product to the ongoing sinusoidal disturbances of the process has been investigated in the film casting of viscoelastic polymer fluids using frequency response analysis. As demonstrated for fiber spinning process (Jung et al., 2002; Devereux and Denn, 1994), this frequency response analysis is useful for examining the process sensitivity and the stability of extensional deformation processes including film casting. The results of the present study reveal that the amplification ratios or gains of the process/product variables such as the cross-sectional area at the take-up to disturbances exhibit resonant peaks along the frequency regime as expected for the systems having hyperbolic characteristics with spilt boundary conditions (Friedly, 1972). The effects on the sensitivity results of two important parameters of film casting, i.e., the fluid viscoelasticity and the aspect ratio of the casting equipment have been scrutinized. It turns out that depending on the extension thinning or thickening nature of the fluid, increasing viscoelasticity results in enlargement or reduction of the sensitivity, respectively. As regards the aspect ratio, it has been found that an optimum value exists making the system least sensitive. The present study also confirms that the frequency response method produces results that corroborate well those by other methods like linear stability Analysis and transient solutions response. (Iyengar and Co, 1996; Silagy et al., 1996; Lee and Hyun, 2001).

• Biomolecules & Therapeutics
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• v.9 no.2
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• pp.79-87
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• 2001

Heat shock proteins serve as chaperone by preventing the aggregation of denatured proteins and promote survival of pathogens in harsh environments. In this study, heat shock gene encoding a 84-kDa (p84) protein, which is one of the three major heat shock proteins in S. pneumoniae, was cloned and characterized. PCR with a forward primer derived from N-terminal amino acid sequence of the p84 and a reverse primer derived from the conserved second ATP-binding region of Clp family was used for amplification of the gene encoding the p84 and subsequently the PCR product was used for sequence determination. Sequence analysis of the p84 gene demonstrated that it is a member of ClpL. The deduced amino acid sequence of pneumococcal ClpL shows homology with other members of the Clp family, and particularly, even in variable leader region, with bovine Clp-like protein and L. lactis ClpL. S. pneumoniae clpL is the smallest clop member (701 amono acids) containing the two conserved ATP-binding regions, and hydrophilic N-terminal variable region of pneu-mococcal Clp ATPase is much shorter than any known Clp ATPases. Histidine tagged ClpL was overexpressed and purified from E. coli. Immunoblot analysis employing antisera raised against pneumococcus p84 demonstrated no cross-reactivity with Clp analog in Eschericha coli, Staphylococcus aureus and human HeLa cells. Preimmunization of mice with ClpL extended mice life partially but did not protect them from death.

• Structural Monitoring and Maintenance
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• v.9 no.2
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• pp.201-220
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• 2022

Lateral vibrations of footbridges may induce synchronization between pedestrians and structure itself, resulting in amplification of such vibrations, a phenomenon identified by lock-in. However, investigations about accelerations and frequencies of the structural movement that are related to the occurrence of synchronization are still incipient. The aim of this paper is to investigate conditions that could lead to avoidance of synchronization among pedestrians themselves and footbridge, expressed in terms of peak acceleration. The focus is on the low acceleration range, employed in some guidelines as a criterion to avoid synchronization. An experimental campaign was carried out, employing a prototype footbridge that was set into oscillatory motion through a pneumatic exciter controlled by a fuzzy system, with controlled frequency and amplitude. Test subjects were then asked to cross the oscillating structure, and accelerations were simultaneously recorded at the structure and at the subject's waist. Pattern and phase differences between these signals were analysed. The results showed that test subjects tended to keep their walking patterns without synchronization induced by the vibration of the structure, for structural peak acceleration values up to 0.18 m/s², when frequencies of oscillation were around 0.8 to 0.9 Hz. On the other hand, for frequencies of oscillation below 0.7 Hz, structural peak accelerations up to 0.30 m/s² did not induce synchronization.