Yao, Zhuang;Meng, Yu;Le, Huong Giang;Lee, Se Jin;Jeon, Hye Sung;Yoo, Ji Yeon;Kim, Hyun-Jin;Kim, Jeong Hwan
Journal of Microbiology and Biotechnology
/
v.30
no.11
/
pp.1720-1728
/
2020
We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40℃, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.
Journal of the Institute of Electronics Engineers of Korea SP
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v.46
no.6
/
pp.102-111
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2009
In this paper, we propose a de-interlacing algorithm that combines a motion compensation (MC) method and the vertical-temporal filter with motion compensation (MC V-T filter) according to motion compensation reliability. The MC method represent one of the best ways of improving the resolution of de-interlaced frames, but it may introduce motion compensation artifacts in regions with incorrect motion information. In these regions, the MC V-T filter that is very robust to motion vector errors can be used to correct motion compensation artifacts. The combination between two methods is controlled by the motion compensation reliability that is measured by analyzing the estimated motion vectors and the results of MC. The motion compensation reliability contains information about motion compensation artifacts of MC results and determines the combination weight according to this information. Therefore, the combination rule of the proposed method is more accurate than those of the conventional methods and it enables the proposed method to provide high quality video sequences without producing any visible artifacts. Experimental results with various test sequences show that the proposed algorithm outperforms conventional algorithms in terms of both visual and numerical criteria.
An, Ji Young;Park, Byung Bae;Byun, Jae Kyung;Cho, Min Seok;Kim, Yong Suk;Han, Si Ho;Kim, Se Bin
Journal of Korean Society of Forest Science
/
v.104
no.1
/
pp.43-49
/
2015
The production of high quality seedlings is a very important phase in silvicultural systems for successful reforestation or restoration. The purpose of this study was to quantitatively measure both growth performances and nutrient responses of Fraxinus rhynchophylla, Pinus densiflora, and Pinus koraiensis seedlings, which are commercially planted in Korea, according to the different types of soil improvement treatments. We applied soil brought (hereafter 'brought'), subsoil inversion (hereafter 'subsoil'), and mixture of brought soil with soil on nursery bed (hereafter 'mixing') in a permanent national nursery. Silt and clay contents were the highest at the subsoil treatment and organic material, soil nitrogen and phosphorus concentrations were the lowest at the brought treatment. The growth of F. rhynchophylla was the lowest at the subsoil treatment, but there were no significant differences among treatments. There were significant differences in only root nutrient concentrations of F. rhynchophylla among treatments: nitrogen, phosphorus, and potassium concentrations were the lowest at the subsoil or brought treatment. Mixing treatment increased N contents with deduction of N concentrations ('dilution') because of more dry weight increase compared with the amount of N uptake. This study suggested mix of brought soil with soil on a nursery bed in a permanently used nursery can economically be an effective technique to improve soil quality.
Debates over the ecological and public health impacts of aerial pesticide sprays are increasing. This is particularly true for controlling Monochamus beetles, which are vector insects of pinewood nematodes. In 2017, adult female orb-web spiders, Trichonephila clavata, were sampled from pine forests in Yangsan-si, Gyeongsangnam-do, Korea, where the aerial pesticide spray, fenitrothion or thiacloprid, was used for several decades. The biological traits of the spiders (body weight, body length, carapace width, and total hind leg length) were compared among treatment sites (no-spray, sprayed three times, and sprayed five times), and differences were observed. The body length, carapace width, and total hind leg length of the spiders in the sprayed areas were significantly shorter than in the no-spray area, but there were no differences between the area sprayed three or five times. These results indicate that repeated exposures to an aerial pesticide spray can alter morphological parameters, which influences population-level fitness. Future studies should monitor the spider long-term responses to pesticides (a direct effect) and prey availability (an indirect effect).
cDNA encoding somatolactin (SL) was obtained by RT-PCR from pituitary glands of rock bream (Oplegnathus fasciatus). The full length cDNA of rock bream somatolactin (rbSL) is 1636 bp long. It contains a 696 bp open reading frame encoding a signal peptide of 24 amino acids (an) and a mature protein of 207 aa. rbSL has seven cysteine residues$(Cys^{5},\; Cys^{15},\; Cys^{42},\; Cys^{65},\; Cys^{181},\; Cys^{198}\; $and $Cys^{206})$ and two potential N-glycosylation sites at positions $Asn^{121}$and $Asn^{153}$. The rbSL shares 61.1∼92.6% amino acid sequence similarities and 63∼92.6% nucleotide sequence identities with other teleost SLs, except for goldfish and channel catfish SL. Amino acid sequence alignment revealed that rbSL has four conserved domains $(A_{SL},\; B_{SL},\; C_{SL}and\; D_{SL})$ common to all SLs. Out of these domains, $(A_{SL},\; B_{SL},\; C_{SL}and\; D_{SL})$, are also conserved in all teleost growth hormones and prolactins. The cDNA of rbSL has been cloned into pET expression vector in order to produce recombinant rbSL in E. coli BL2l(DE3) cells. The recombinant protein showed a molecular weight of 27 kDa in SDS-PAGE.
A gene for cellulolytic xylanase of Bacillus circulnns ATCC21365 was cloned on pUC 19 in Eschwichia coli. The recombinant plasniid pXLI80 contained an 1.8 id, inselt composed of0.5 kb and 1.3 kb PslI fragments derived from B, circulans. The 0.5 kh fragment in the upstream region of 1.3 kb one was confirmed lo be indispensable for not only expression but also hyperexpression of the cloned gene. The transformant overproduced the xylanase 135 times greater than that produced by the orlginal B circulnns. The optimum pH and temperature of the cloned enzyme we]-e pH 5.2 and $60^{\circ}C$, respectively. Heal pretl-eatment at TEX>$55^{\circ}C$C for 1 Indid not cause inhibition of the activity of this enzyme. The elm.ynie could hydl-olyre CMC and lichenan as well as xylan to produce xylose(or GI), xylohiose(or G2) and xylolnose(or G3) as inah products. Hence We defined the cloned enzyme as a cellulolytic xylanase. The SDS-PAG electrophoretic mobility and zyiiogram of this enzyme derived from whole cell extracts or c~~lture supematants or E. coli(pXL180) indicated a molecular weight of 45,000 and nonprocessing of the enzyme in the peilplasln of E. coli.
Proceedings of the Korean Society of Life Science Conference
/
2001.06a
/
pp.67-86
/
2001
A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.
The amino acid analysis of polyhedrin protein and nucleotide sequence of polyhedrin gene in H. cunea nuclear polyhedrosis virus (HcNPV) genome have been studied. Polyhedrin had three polypeptide bands in SDS - polyactylamide gel electrophoresis. The major polypeptide had a molecular weight of 25 kd. The polyhedrin was composed of 17 different amino acids. HcNPV DNA was digested with EcoRI restriction enzyme and hybridized with ($\alpha^{32}P$) -labelled AcNPV polyhedrin gene cDNA. The polyhedrin gene was located on the fragment of EcoRI-H. The EcoRI - H fragment containing polyhedrin gene was cloned into the EcoRI site of pUC8 vector which was confirmed with southern blotting, and the recombinant plasmid containg polyhedrin gene was designated as hPE-H. The promoter region of polyhedrin genomic DNA was sequenced. The sequences identified as the TATA box was found at the 5' flanking region of the polyhedrin genomic DNA approximately -79 bp upstream from the transcriptional start site. But CAAT-like box was not shown near the TATA-like box in the polyhedrin gene. Four tandem repeats with the sequence 5' -CTAATAT-3' and 5'-TAAATAA-3' were found between -141 and -108 or -83 upstream and -52 bp downstream from the translation start site. About -141 bp region upstream from the translational start site was highly AT (78%) rich. The coding region for the polyhedrin starts and ends with ATG and TAA, respectively.
Speech articulators are coordinated for the purpose of segmental constriction in terms of a task. In particular, vertical jaw movements repeatedly contribute to consonantal as well as vocalic constriction. The current study explores vertical jaw movements in conjunction with bilabial constriction in bilabial stop /p/ in the context /a/-to-/a/. Revisiting kinematic data of /p/ collected using the electromagenetic midsagittal articulometer (EMMA) method from seven (four female and three male) speakers of Seoul Korean, we examined maximum vertical jaw position, its relative timing with respect to the upper and lower lips, and lip aperture minima. The results of those dependent variables are recapitulated in terms of linguistic (different word boundaries) and paralinguistic (different speech rates) factors as follows. Firstly, maximum jaw height was lower in the across-word boundary condition (across-word < within-word), but it did not differ as a function of different speech rates (comfortable = fast). Secondly, more reduction in the lip aperture (LA) gesture occurred in fast rate, while word-boundary effects were absent. Thirdly, jaw raising was still in progress after the lips' positional extrema were achieved in the within-word condition, while the former was completed before the latter in the across-word condition. Lastly, relative temporal lags between the jaw and the lips (UL and LL) were more synchronous in fast rate, compared to comfortable rate. When these results are considered together, it is possible to posit that speakers are not tolerant of lenition to the extent that it is potentially realized as a labial approximant in either word-boundary condition while jaw height still manifested lower jaw position in the across-word boundary condition. Early termination of vertical jaw maxima before vertical lower lip maxima across-word condition may be partly responsible for the spatial reduction of jaw raising movements. This may come about as a consequence of an excessive number of factors (e.g., upper lip height (UH), lower lip height (LH), jaw angle (JA)) for the representation of a vector with two degrees of freedom (x, y) engaged in a gesture-based task (e.g., lip aperture (LA)). In the task-dynamic application toolkit, the jaw angle parameter can be assigned numerical values for greater weight in the across-word boundary condition, which in turn gives rise to lower jaw position. Speech rate-dependent spatial reduction in lip aperture may be able to be resolved by means of manipulating activation time of an active tract variable in the gestural score level.
The aim of this research was to develop a climate change vulnerability index at the district level (Si, Gun, Gu) with respect to the health care sector in Korea. The climate change vulnerability index was esimated based on the four major causes of climate-related illnesses : vector, flood, heat waves, and air pollution/allergies. The vulnerability assessment framework consists of six layers, all of which are based on the IPCC vulnerability concepts (exposure, sensitivity, and adaptive capacity) and the pathway of direct and indirect impacts of climate change modulators on health. We collected proxy variables based on the conceptual framework of climate change vulnerability. Data were standardized using the min-max normalization method. We applied the analytic hierarchy process (AHP) weight and aggregated the variables using the non-compensatory multi-criteria approach. To verify the index, sensitivity analysis was conducted by using another aggregation method (geometric transformation method, which was applied to the index of multiple deprivation in the UK) and weight, calculated by the Budget Allocation method. The results showed that it would be possible to identify the vulnerable areas by applying the developed climate change vulnerability assessment index. The climate change vulnerability index could then be used as a valuable tool in setting climate change adaptation policies in the health care sector.
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