• International Journal of Industrial Entomology and Biomaterials
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• v.8 no.2
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• pp.145-149
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• 2004

The expression of a fusion protein comprised of the B. thuringiensis crystal protein, Cry1Ac, and enhanced green fluorescent protein (EGFP) in Escherichia coli XLl-blue was examined. Three recombinant plasmids were transformed into E. coli XL1-blue and named as ProAc/Ec, MuEGFP/Ec and ProMu-EGFP/Ec, respectively. All transformants were observed by light and fluorescence microscopy at mid-log phase. The expression in E. coli transformants, ProMu-EGFP/Ec and MuEGFP/Ec, exhibited bright enough fluorescence to be observed. Furthermore, ProMu-EGFP/Ec produced fluorescent inclusions, which may have been recombinant crystals between EGFP and Cry1Ac while MuEGFP/Ec expressed soluble EGFP in cell. In SDS-PAGE, ProAc/Ec had 130 kDa crystal protein band and MuEGFP/Ec had thick 27 kDa EGFP band. However, ProMu-EGFP/Ec had about 150 kDa fusion protein band. Accordingly, these results indicated that a fusion protein between the B. thuringiensis crystal protein and a foreign protein under the lacZ promoter was successfully expressed as granular structure in E. coli. It is suggested that the E. coli expression system by N-terminal fusion of B. thuringiensis crystal protein may be useful as excellent means for fusion expression and characterization of B. thuringiensis fusion crystal protein.

• International Journal of Industrial Entomology and Biomaterials
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• v.8 no.1
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• pp.89-93
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• 2004

Expression of a fusion protein between B. thuringiensis crystal protein, Cry1Ac1 and a scorpion insect toxin (AaIT, Androctonus australis Hector insect toxin) in acrystalliferous B. thuringiensis strain (Cry-B strain) was examined. The cry 1Ac1 gene was cloned in B. thuringiensis-E coli shuttle vector, pHT3101, under the control of the native cry 1Ac1 gene promoter (pProAc) and a gene encoding AaIT was inserted in XhoI site in the middle of the cry 1Ac1 gene (pProAc-ScoR). B. thuringiensis Cry-B strain carrying pProAc-ScoR (PyoAc-ScoR/CB) produced an inclusion body of irregular shape and the expressed fusion protein is approximately 65 kDa in size. Sporulated cells and spore-crystal mixtures of ProAc-ScoR/CB had insecticidal activity against Plutella xylostella larvae, showing $LT_50$ of ProAc-ScoR/CB (22.59 hrs) lower than that of ProAc/CB (30.06 hrs) at $1{\times}{10^7} {CEU/cm^2}$. These results suggest that the fusion protein including a B. thuringiensis crystal protein and an AaIT may be functionally expressed in B. thupingiensis. Moreover, we verified the additive toxicity of AaIT, which is a new feasible candidate for insect control.

• Journal of Microbiology
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• v.42 no.4
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• pp.340-345
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• 2004

To investigate the co-expression and crystallization of a fusion gene between the Bacillus thuringiensis crystal protein and a foreign protein in B. thuringiensis, the expression of the Cry1Ac fused with green fluorescent protein (GFP) genes in a B. thuringiensis $Cry^-B$ strain was examined. The cry1Ac gene was cloned in the B. thuringiensis-E. coli shuttle vector, pHT3101, under the control of the native cry1Ac gene promoter, while the GFP gene was inserted into the XhoI site upstream of the proteolytic cleavage site, in the middle region of the crylAc gene (pProAc-GFP). The B. thuringiensis $Cry^-B$ strain carrying pProAc-GFP (ProAc-GFP/CB) did not produce any inclusion bodies. However, the transformed strain expressed fusion protein forms although the expression level was relatively low. Furthermore, an immu­noblot analysis using GFP and Cry1Ac antibodies showed that the fusion protein was not a single spe­cies, but rather multiple forms. In addition, the N-terminal fragment of Cry1Ac and a non-fused GFP were also found in the B. thuringiensis $Cry^-B$ strain after autolysis. The sporulated cells before autolysis and the spore-crystal mixture after autolysis of ProAc-GFP/CB exhibited insecticidal activities against Plutella xylostella larvae. Accordingly, the current results suggest that a fusion crystal protein produced by the transfomant, ProAc-GFP/CB, can be functionally expressed but easily degraded in B. thuring­iensis.

• BMB Reports
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• v.40 no.3
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• pp.396-403
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• 2007

Proline accumulation has been shown to correlate with tolerance to drought and salt stresses in plants. We attempt to introduce the wild-type, mutant, and fusion proBA genes derived from Bacillus subtilis into Arabidopsis thaliana under the control of a strong promoter cauliflower mosaic virus 35S (CaMV35S). The transgenic plants produced higher level of free proline than control and the overproduction of proline resulted in the increased tolerance to osmotic stress in transgenic plants. Besides, the mutation in proBA genes, which were proved to lead $\alpha$-glutamyl kinase ($\alpha$-GK) reduces sensitivity to the end-product inhibition and the fusion of proB and proA also result in increasing proline production and confer osmotolerance in transgenic lines.

• Journal of the Korean Society for Precision Engineering
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• v.24 no.8 s.197
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• pp.122-129
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• 2007

To evaluate the effect of artificial disc implantation and fusion on the biomechanics of adjacent motion segment, a nonlinear three-dimensional finite element model of whole lumbar spine (L1-S1) was developed. Biomechanical analysis was performed for two different types of artificial disc, ProDisc and SB $Charit{\acute{e}}$ III model, inserted at L4-L5 level and these results were also compared with fusion case. Angular motion of vertebral body, forces on the spinal ligaments and facet joint under sagittal plane loading with a compressive preload of 150 N at a nonlinear three-dimensional finite element model of Ll-S1 were compared. The implant did not significantly alter the kinematics of the motion segment adjacent to the instrumented level. However, $Charit{\acute{e}}$ III model tend to decrease its motion on the adjacent levels, especially in extension motion. Contrast to motion and ligament force changes, facet contact forces were increased in the adjacent levels as well as implanted level for constrained instantaneous center of rotation model, i.e. ProDisc model.

• Journal of Fisheries and Marine Sciences Education
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• v.28 no.4
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• pp.903-912
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• 2016

Hagfish which belongs to the chordate contact cyclostomata, is important phylogenetic relationship between vertebrate and invertebrate. Cathepsins of the cysteine protease family have traditionally been thought to play a major role in intracellular protein degradation and turnover in lysosomes. In this study, Catepsin L was cloned from Inshore hagfish (Eptatretus burgeri), the cDNA encoding ORF of the Eptatretus burgeri Cathepsin L (EbCtL) is 978 bp. The cDNA encoding proEbCtL was expressed in Escherichia coli strain BL21(DE3) using the pGEX-4T-1 expression vector system. The recombinant proEbCtL protein was overexpressed as a approximately 55 kDa fusion protein. The overproduced soluble GST-fusion protein was then applied to glutathione-Sepharose 4B column chromatography; the sample harboring the fusion protein evidenced a high degree of purity when analyzed via SDS-PAGE and Western blot analysis. Its activity was quantied by cleaving the synthetic peptide Z-FR-AMC, Z-LLE-AMC, and Suc-AAF-AMC, and the optimal pH for the protease activity was 8, 9.5, and 9, respectively.

• Bulletin of the Korean Chemical Society
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• v.30 no.5
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• pp.1207-1210
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• 2009

• Current Optics and Photonics
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• v.7 no.3
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• pp.237-247
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• 2023

In middle- and long-distance imaging systems, due to the atmospheric turbulence caused by temperature, wind speed, humidity, and so on, light waves propagating in the air are distorted, resulting in image-quality degradation such as geometric deformation and fuzziness. In remote sensing, astronomical observation, and traffic monitoring, image information loss due to degradation causes huge losses, so effective restoration of degraded images is very important. To restore images degraded by atmospheric turbulence, an image-restoration method based on improved compound multibranch feature fusion (CMFNetPro) was proposed. Based on the CMFNet network, an efficient channel-attention mechanism was used to replace the channel-attention mechanism to improve image quality and network efficiency. In the experiment, two-dimensional random distortion vector fields were used to construct two turbulent datasets with different degrees of distortion, based on the Google Landmarks Dataset v2 dataset. The experimental results showed that compared to the CMFNet, DeblurGAN-v2, and MIMO-UNet models, the proposed CMFNetPro network achieves better performance in both quality and training cost of turbulent-image restoration. In the mixed training, CMFNetPro was 1.2391 dB (weak turbulence), 0.8602 dB (strong turbulence) respectively higher in terms of peak signal-to-noise ratio and 0.0015 (weak turbulence), 0.0136 (strong turbulence) respectively higher in terms of structure similarity compared to CMFNet. CMFNetPro was 14.4 hours faster compared to the CMFNet. This provides a feasible scheme for turbulent-image restoration based on deep learning.

• The Korean Journal of Mycology
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• v.21 no.3
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• pp.200-211
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• 1993

Somatic hybrids of Pleurotus florida ASI 2016 and Pleurotus ostreatus ASI 2018 were obtained by protoplast fusion. The 40 fusants($P1{\sim}P40$) was examined for the yield on fermented and pasteurized rice straw in a tray. The carpophore yield of them were showed as the range of $27.0{\sim}155.2$, based on parental values of 100(ASI 2018), The pilei of fusants between orange white colored P. florida and dark grey colored P. ostreatus had mixed colors in the young stage. Other breeding programmes were performed to improve new varieties with high yield and good quality. A new oyster mushroom variety, Wonhyeongneutaribeosus(P72), was developed at the Agricultural Sciences Institute, Rural Development Administration in 1990. This P.florida-ostreatus-ostreatus hybrid P72 was selected from 38 protoplast fusion products($P41{\sim}P78$) between P.florida-ostreatus recombinant P5-M 43-arg rib and P. ostreatus ASI 2-13-0 2001-19-pro orn. The yield indexes of 38 hybrids ranged $40.5{\sim}152.7$ compared with the parental values of 100(ASI 2001). Hybrid P72 was characterized by the large fruiting bundle of semispherical shape with long stipe and by the small and circular pileus, resulting in lower harvesting cost. A significant increase in carpophore production was observed in somatic hybrids of protoplasts due to heterosis. A comparision of hybrid with parents P72 was made using isozyme analysis. The esterase banding patterns could be characterized by new bands in the hybrids. Seven fusion products of four crosses between P.florida ASI 2016 and P. ostreatus ASI 2018 were analysed with respect to the distribution of progenies and segregation of gene markers by random basidiospore analysis. Segregation of alleles should yield progeny of four genotypes in a Mendelian ratio of 1 : 1 : 1 : 1 for prototrophs, auxotrophs of one parental type, auxotrophs of the other parental type, and auxotrophic recombinants, respectively. However, five fusants of them did not detect one parental, P.ostreatus, type. Basidiospores could yield progeny of 16 genotypes in the cross of one of the recombinant P5-M43-arg $rib{\times}P. ostreatus$ ASI 2-13-pro orn but the segregants of three fusants were not detected clearly. The allele ratio of loci could be expected 1 : 1 : 1 : 1 for arg, rib, pro and orn. The ratio, however, would be changed to 4 : 1 : 1 : 1 with increasing proportion of argo In almost all the fusants, prototrophic recombinants were recovered in large numbers against auxotrophic markers. Parental genotypes were recovered with the recombinant progeny amounting to $38.68{\sim}99.56%$. The analysis provides proof of heterokaryosis and strong evidence for haploidy of vegetative nuclei, a sexual cycle consisting of nuclear fusion and meiosis.

• Proceedings of the Korean Nuclear Society Conference
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• 2004.10a
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• pp.719-720
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• 2004

A multi-megawatt long pulse ion source (LPIS) of neutral beam injector was developed for the KSTAR. Beam extraction experiments of the LPIS were carried out at the neutral beam test stand (NBTS). Design requirements for the ion source were 120 kV/65 A deuterium beam and a 300 s pulse length. A maximum ion density of $9.1310^{11}$ $cm^{-3}$ was measured by using electric probes, and an optimum arc efficiency of 0.46 A/kW was estimated with ion saturation current of the probes, arc power, and total beam area. An arcing problem, caused by the structural defect of decelerating grid supporter, in the third gap was solved by the blocking of backstream ion particles, originated from the plasma in the neutralizer duct, through the unnecessary spaces on the side of grid supporter. A maximum drain power of 1.5 MW (i.e. 70 kV/21 A) with hydrogen was measured for a pulse duration of 0.5 s. Optimum beam perveance was ranged from 0.75 to 0.85. An improved design of accelerator for the effective control of beam particle trajectory should provide higher beam perveance.