• The Korean Journal of Mycology
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• v.26 no.4 s.87
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• pp.450-457
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• 1998

The cryparin of Cryphonectria parasitica belongs to a cell wall associated fungal hydrophobin. The cryparin, though it is encoded by a single copy gene, is known for the high expression during the liquid culture of C. parasitica, and it turns out that 22% of total mRNA was transcribed for cryparin at 48hr after the liquid culture. In addition, it is also known as one of down-regulated fungal proteins by the presence of double stranded RNA virus, Cryphonectria hypovirus 1. In previous studies (Kim et al., 1999), we have constructed a cryparin-null mutant by replacing the cryparin gene with hygromycin B resistance gene due to site directed homologous recombination. In order for the promoter analysis of cryparin which seems to be very strong as well as mycoviral specific, it is preferable to have a strain with only a target promoter replaced and a discernable target site for incoming vectors. However, the cryparin-null mutant revealed the presence of an additional copy of transforming vector except the one which replaced the cryparin gene. In addition, the cryparin-null mutant did not contain any markers for targeted integration of incoming vectors. This prompts us to design an experiment to obtain a strain for promoter analysis of cryparin gene. A different mating type strain EP6(Mata, $met^-$) was mated with the cryparin-null mutant ${\triangle}$Crp194-7(MatA, Crp${\triangle}$::hph) to make the progenies with only a single replacement vector and $met^-$ characteristic remained. Nutritional assay as well as Southern blot analysis revealed that the progeny, ${\triangle}$Crp194-a6, was the methionine auxotroph with a single replacing vector in genome. Northern blot analysis and PAGE showed that there was no cryparin produced in this bred strain either.

• The Journal of Korean Society of Virology
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• v.30 no.1
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• pp.71-81
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• 2000

We have determined and analyzed the full-length cDNA sequence of a coxsackievirus B3 (CVB3) Korean isolate (CVB3-Korea/97) which has been known as a general human pathogen. The whole genome contains 7,400 nucleotides and has a single large open reading frame with 6,555 nucleotides that encodes a potential polyprotein precursor of 2,185 amino acids. The genome also contains a 5' non-coding region (NCR) of 741 bases and a 3' NCR of 104 bases followed by poly(A) tail. Sequence homologies of nucleotides and deduced amino acids between the CVB3-Korea/97 strain and the prototype (Nancy strain) were 81.7% and 91.5%, respectively. The genes encoding the functional proteins including viral protease and RNA dependent RNA polymerase showed higher homology than those encoding the structural proteins. We have further analyzed the sequences of 5' NCR, VP1 and VP2 of CVB3-Korea/97, which are known as cardiovirulent determining factors at the nucleotide and amino acid levels. Although the CVB 3-Korea/97 strain was isolated from an aseptic meningitis patient without cardiomyopathy, its 234th nucleotide and 165th amino acid were uracil and Asn as same as those of other cardiovirulent strains one. However, the 155th amino acid of VP1, which closely associated with cardiovirulence, was replaced with $Arg^{155}$ by single nucleotide substitution from $A^{2916}$ to $T^{2916}$. Moreover, additional amino acid substitutions were observed in the flanking region of $Asp^{155}$. Taken together, amino acid(s) substitution in VP1 may playa critical role in determining cardiovirulence of the CVB3-Korea/97 strain rather than individual nucleotide replacements in the 5' NCR and/or an amino acid substitution in VP2.

• Journal of the Korean GEO-environmental Society
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• v.12 no.11
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• pp.71-77
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• 2011

Recently, non-destructive stress measurement method using magnetic anisotropy sensor has been applied to the construction site such as steel bridges and steel pipes. In addition, steel rib used in the tunnel construction site was found to be possible to measure the stress by non-destructive method. In this study, steel loading experiments using magnetic anisotropy sensor developed in Japan and strain gauges were conducted to derive stress sensitivity curve for domestic steel SS400. Also, additional steel loading experiments and numerical analysis were performed for evaluation of applicability for non-destructive stress measurement method using magnetic anisotropy sensor. As a result of this study, stress sensitivity curves for domestic steel SS400 were derived using output voltage measured by magnetic anisotropy sensor and average of stress measured by strain gauges depending on the measurement location. And as a result of comparing additional steel loading experiments with the numerical analysis, error level of magnetic anisotropy sensor is around 20MPa. When considering the level of the yield stress(245MPa) of steel, in case of using magnetic anisotropy sensor in order to determine the stress status of steel, it has sufficient accuracy in engineering. Especially, magnetic anisotropy sensor can easily identify the current state of stress which considers residual stress at steel structure that stress measurement sensor is not installed, so we found that magnetic anisotropy sensor can be applied at maintenance of steel structure conveniently.

• Journal of the Computational Structural Engineering Institute of Korea
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• v.16 no.2
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• pp.165-172
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• 2003

In this study, a new and efficient harmonic axisymmetric shell element for static and dynamic analysis Is proposed. The present element considering shear strain is based on a modified mixed variational principle in which the independent unknowns are only the Quantities prescribable at the shell edges. Unlike existing hybrid-mixed axisymmetric shell elements, the present element introduces additional nodeless degrees for displacement field Interpolation In order to enhance the numerical performance. The stress parameters are eliminated by the stationary condition and the nodeless degrees are condensed out by the Guyan reduction. Through several numerical examples, the hybrid-miked shell element with the additional nodeless degrees and the consistent stress parameters is shown to be efficient and yield very accurate results for static and vibration analysis.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2004.10a
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• pp.756-759
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• 2004

We present a piezoelectric actuator using stiffness control and stroke amplification mechanism in order to make large lateral displacement. In this work, we suggest stiffness control approach that generates lateral displacement by increasing the vertical stiffness and reducing the lateral stiffness using additional structure. In addition, an additional structure of a serpentine spring amplifies the lateral displacement like leverage structure. The suggested lateral PZT actuator (bellows actuator) consists of serpentine spring and PZT/electrode layer which is located at the edge of the serpentine spring. The edge of the serpentine spring prevents the vertical motion of PZT layer, while the other edge of the serpentine spring makes stroke amplification like leverage structure. We have determined dimensions of the bellows actuator using ANSYS simulation. Length, width and thickness of PZT layer are 135$\mu$m, 20$\mu$m and 0.4$\mu$m, respectively. Dimensions of the silicon serpentine spring are thickness of 25$\mu$m, length of 300$\mu$m, and width of 5$\mu$m. The bellows actuator has been fabricated by SOI wafer with 25$\mu$m-top silicon and 1$\mu$m-buried oxide layer. The bellows actuator shows the maximum 3.93$\pm$0.2$\mu$m lateral displacement at 16V with 1Hz sinusoidal voltage input. In the frequency response test, the fabricated bellows actuator showed consistent displacement from 1Hz to 1kHz at 10V. From experimental study, we found the bellows actuator using thin film PZT and silicon serpentine spring generated mainly laterally displacement not vertical displacement at 16V, and serpentine spring played role of stroke amplification.

• Asian Journal of Atmospheric Environment
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• v.5 no.3
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• pp.164-171
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• 2011

This work focuses on the analysis of bioaerosols in the atmosphere at higher altitudes over Noto Peninsula, Japan. We carried out direct sampling via aircraft, separated cultures, and identified present isolates. Atmospheric bioaerosols at higher altitudes were collected using a Cessna 404 aircraft for an hour at an altitude of 3,500 m over the Noto Peninsula. The aircraft-based direct sampling system was devised to improve upon the system of balloon-based sampling. In order to examine pre-existing microorganism contamination on the surface of the aircraft body, bioaerosol sampling was carried out just before takeoff using the same method as atmospheric sampling. Identification was carried out by a homology search for 16S or 18S rDNA isolate sequences in DNA databases (GenBank). Isolate sampling just before takeoff revealed Stretpomyces sp., Micrococcus sp., and Cladosporium sp. One additional strain, Bacillus sp., was isolated from the sample after bioaerosol collection at high altitude. As the microorganism contamination on the aircraft body before takeoff differed from that while in the air, the presence of additional, higher atmosphere-based microorganisms was confirmed. It was found that Bacillus sp. was floating at an altitude of 3,500 m over Noto Peninsula.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1125-1129
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• 2005

Cloning of a 6.6-kb BamHI digested chromosomal DNA from S. griseus IFO13350 revealed the presence of an additional gene encoding a novel trypsin-like enzyme, named SprU. The SprU protein shows a high homology ($79\%$ identity, $88\%$ similarity) with the SGT protease, which has been reported as a bacterial trypsin in the same strain. The amino acid sequence deduced from the nucleotide sequence of the sprU gene suggests that SprU is produced as a precursor consisting of an amino-terminal presequence (29 amino acid residues), prosequence (4 residues), and mature trypsin consisting of 222 amino acids with a molecular weight of 22.94 kDa and a calculated pI of 4.13. The serine, histidine, and aspartic acid residues composing the catalytic triad of typical serine proteases are also well conserved. When the trypsin activity of the SprU was spectrophotometrically measured by the enzymatic hydrolysis of the artificial chromogenic substrate, N-${alpha}$-benzoyl-DL-arginine-p-nitroanilide, the S. lividans transformant with pWHM3-U gave 3 times higher activity than that of control. When the same recombinant plasmid was introduced into S. griseus, however, the gene dosage effect was not so significant, as in the cases of other genes encoding serine proteases, such as sprA, sprB, and sprD. Although two trypsins, SprU and SGT, have a high degree of homology, the pI values, the gene dosage effect in S. griseus, and the gene arrangement adjacent to the two genes are very different, suggesting that the biochemical and biological function of the SprU might be quite different from that of the SGT.

• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.918-929
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• 2015

Microbial enhanced oil recovery (MEOR) is being used more widely, and the biological contributions involved in MEOR need to be identified and quantified for the improvement of field applications. Owing to the excellent interfacial activity and the wide distribution of producing strains in oil reservoirs, lipopeptides have proved to be an essential part of the complex mechanisms in MEOR. In this study, crude lipopeptides were produced by a strain isolated from an indigenous community in an oil reservoir. It was found that crude lipopeptides can effectively reduce the IFT (interfacial tension) to 10^-1~10^-2 mN/m under high salinity without forming stable emulsions, and the wettability of natural sandstone can be enhanced (Amott index, from 0.36 to 0.48). The results of core flooding experiments indicate that an additional 5.2% of original oil in place can be recovered with a 9.5% reduction of injection pressure. After the shut-in period, the wettability of the core, the reduction of injection pressure, and the oil recovery can be improved to 0.63, 16.2% and 9.6%, respectively. In the microscopic flooding experiments, the crude oil in membrane, cluster, and throat states contribute nearly 90% in total of the additional oil recovery, and the recovery of membranestate oil was significantly enhanced by 93.3% after shut in. Based on the results in macro and pore scale, the IFT reduction and the wettability alteration are considered primary contributors to oil recovery, while the latter was more dominant after one shut-in period.

• Bulletin of the Korean Chemical Society
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• v.28 no.12
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• pp.2303-2309
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• 2007

Expressed protein ligation (EPL) technique, joining recombinantly expressed proteins to polypeptides, has been widely adopted for addressing various biological questions and for drug discovery. However, joining two recombinant proteins together is sometimes difficult when proteins are expressed insoluble and unrefoldable, because ligation-active proteins via intein-fusion are obtainable when they are folded correctly. We overcame this limitation coexpressing target protein with additional methionine aminopeptidase (MAP) which enhances removal of the initiation methionine of recombinantly expressed protein. Our approach demonstrated that two domains of 46 kDa 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, a target of herbicide glyphosate, were successfully joined by native chemical ligation, although its C-terminal domain was expressed as an inclusion body. The intein-fused N-terminal fragment of EPSP synthase (EPSPSN, residues 1-237) was expressed and the ligation-active thioester tagged N-terminal fragment (EPSPSN-thioester) was purified using a chitin affinity chromatography and mercapto-ethanesulphonate (MESNA) as intein thiolysis reagent. Its Cterminal fragment (EPSPSC, residues Met237-238CYS-427), expressed as an inclusion body, was prepared from an additional MAP-expressing strain. Protein ligation was initiated by mixing ~1 mM of EPSPSN-thioester with ~2 mM of EPSPSCCYS (residues 238CYS-427). Also we found that addition of 2% thiophenol increased the ligation efficiency via thiol exchange. The ligation efficiency was ~85%. The ligated full-length EPSP synthase was dissolved in 6 M GdHCl and refolded. Circular dichroism (CD) and enzyme activity assay of the purified protein showed that the ligated enzyme has distinct secondary structure and ~115% specific activity compared to those of wild-type EPSP synthase. This work demonstrates rare example of EPL between two recombinantly expressed proteins and also provides hands-on protein engineering protocol for large proteins.

• Transactions of Materials Processing
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• v.17 no.2
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• pp.124-129
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• 2008

Net shape forging technologies give many effects into the costs and qualities for the finished products. So, the studies to reduce the additional machining amount are very important in forging industry. Specially, there are two main topics in cold forging industry, such as, tool life and precision forging. In this study, new forging technique was proposed to eliminate the machining process for fixing up the length and improve the lead accuracy of gear. The luck-up hub is manufactured through many processes, such as upsetting, piercing and direct extrusion. The gear is formed in direct extrusion process; however, lead accuracy of the gear is over allowance limit. Therefore, the additional sizing process must be added. In this study, process design for closed-die forging of a lock-up hub used for a component of automobile transmission was made using three-dimensional finite element simulations, and the strain distributions and velocity distributions are investigated through the post processor. The rigid-plastic finite-element method for back pressure forging has been used in order to reduce development time and die cost. Using the FEM simulation, we found the optimum value of back pressure. The prototypes of lock-up hub parts were forged into the net-shape. In the experiment, lead precision of tooth are measured by the CCMM(Contact Coordinate Measuring Machine). The dimensional accuracy of forged part was improved up to the 40% when back press was applied.