• Journal of Life Science
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• v.13 no.4
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• pp.463-472
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• 2003

Interactions between factor Va (HFVa) and membrane phosphatidylserine (PS) regulate the activity of the prothrombinase complex. I have previously shown that two solvent exposed hydrophobic residues located in the C2-domain, Trp2063 and Trp2064, are required for binding to immobilized PS and for expression of procoagulant activity on membranes containing 5% PS. In order to fully define the functional importance of these two residues I have expressed and isolated recombinant factor Va (rHFVa) W2063A/W2064A double mutant. In contrast to the native protein the two glycoforms resulting from alternative glycosylation of Asn2181 eluted as a single peak with rHFVa1 W2063A/W2064A eluting on the leading edge and rHFVa2 W2063A/W2064A eluting on the trailing edge. The double mutant rHFVa2 W2063A/W2064A expressed little or no procoagulant activity on membranes containing 1-10% mol % PS. In contrast, the procoagulant activity of this mutant was slightly greater than the native protein on membranes containing>18 mol % PS. The binding of rHFVa2 W2063A/W2064A to immobilized phospholipid vesicles was markedly reduced compared to the native protein in a surface plasmon resonance binding assay. I conclude that Trp2063 and Trp2064 are required for high affinity binding of factor Va to PS membranes and that this interaction is necessary for assembly of the prothrombinase complex on membranes containing physiological concentrations of PS.

• Journal of the Korea Society of Computer and Information
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• v.13 no.3
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• pp.35-44
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• 2008

In this paper, we present an EST sequence annotation system based on Service Oriented Architecture, called SeqWeB. We developed the web services of eight applications (Phred, cross_match, RepeatMasker, TGICL, ICAtools, CAP3, Phrap and Blast) which are located in sequence annotation process and integrated the web services through BFEL. SeqWeB uses an XML file format for data input and output to maximize interoperability between each application. SeqWeB can be extended or modified easily through some modification such as insertion, deletion and replacement because service-oriented architecture allows loose coupling between applications.

• 한국사회정책
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• v.25 no.1
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• pp.345-375
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• 2018

This study examines labour market and unemployment protection policies as a configuration in 12 OECD countries in order to investigate how countries from different regime conform to or diverse from previous welfare state regime discussion, and to examine its relationship with poverty and inequality. In analyzing the combination of the unemployment insurance, the unemployment assistance, and active labour market policy, firstly, fuzzy scores of unemployment insurance was calculated by analyzing the strictness of eligibility, duration of benefit and the generosity of income replacement rate. For unemployment assistance, the ratio of public assistance expenditure to the GDP in each country and the ratio of unemployment benefit level to the average wage in each country have been considered. As for the active labour market policy, the total expenditure per GDP of this policy was converted into fuzzy points and analyzed. As a result, 5 types in 2005 and 6 types in 2010 were generated. Specifically, 'assistance type(iAp)', 'insurance type (Iap)', 'comprehensive safety net type (IAP)', 'weak safety net type(iap)' were analyzed. This paper suggested policy implication for South Korean case, which consistently had high score for weak safety net type(iap).

• Journal of the Korean Recycled Construction Resources Institute
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• v.5 no.4
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• pp.389-394
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• 2017

The aim of the research is providing the application method of recycled materials to manufacture the low costed eco-friend block at currently operated concrete block plant. In this research, based on the previous research results on three types of slag cement with illite, desulfurized gypsum, and wasted refractory products, the actual block product was manufactured by the currently operated plant facility and evaluated their properties to suggest the optimal proportions. As an experimental results, in aspect of compressive strength, absorption ratio, freezing resistance, and pH, type III slag incorporating 5% desulfurized gypsum with 1% replaced illite as an aggregate could be suggested as am optimal proportion. In additionally, considering the high cost of the illite, it can be considered as an optimal proportion that type III slag incorporating 5% desulfurized gypsum for binder.

• Korean Chemical Engineering Research
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• v.55 no.1
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• pp.27-33
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• 2017

The determination of disulfide bonds is important for comprehensive understanding of the chemical structure of protein. So far, many strategies for the disulfide bond analysis have been suggested in terms of speed and sensitivity. However, most of these strategies have not considered free thiol residues in the target protein in the process of determining the disulfide bond. We suggested the strategy which was composed of four steps for the identification of disulfide bonds; the first step was the prediction of possible disulfide bonds, the second step was the determination of free cysteine residues, the third step was the analysis of disulfide bond using a high-resolution mass spectrometry, and the final step was the determination of disulfide bonds based on the comprehensive verification. In this study, we performed the characterization of disulfide bonds for the recombinant protein (HRPE1), where 1 and 5 inter- and intra-chain disulfide bonds were identified, respectively.

• Journal of Life Science
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• v.15 no.5 s.72
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• pp.734-738
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• 2005

A gene coding for xylanase from alkali-tolerant Bacillus alcalophilus AX2000 was cloned into Escherichia coli $DH5\alpha$ using pUC19. Among 2,000 transformants, one transformant showed clear zone on the detection agar plate containing oat-spells xylan. Its recombinant plasmid, named pXTY99, was found to carry 7.0 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynT) was determined, xynT gene was found to consist of 1,020 base-pair open reading frame coding for a poly-peptide of 340 amino acids with a deduced molecular weight of 40 kDa. The coding sequence was preceded by a putative ribosome binding site, and the transcription initiation signals. The deduced amino acid sequence of xylanase is similar to those of the xylanases from Bacillus sp. Nl37 and B. stearothermophilus 21 with $61\%$ and $59\%$ identical residues, respectively.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.248-256
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• 1996

The present study was performed to clone and express human poly (ADP-ribose) synthetase (PARS) cDNA in E coli. For these purposes, the CDNA for human poly (ADP-ribose) synthetase, encoding the entire protein, was cloned into pGEM-7Zf(+). The resulting recombinant plasmid pPARS6.1 was restriction enzyme mapped and its identity was confirmed by Southern blot analysis. The pPARS6. 1 contained full-length CDNA of human PARS and the nudeotide sequences were identical with those reported previously. The recombinant protein which migrated as a unique 120 kDa band on 10% SDS-polyacrylamide gels, was identified as PARS by Southwestern blots using nick-translated DNA probes and by activity gels and activity blots using 32 P-NAD as a substrate for poly (ADP-ribose) synthetase (PARS). The signals corresponding to 120 and 98 kDa proteins were obtained following IPTG (0.4 mM) induction of the PARS cDNA cloned into Xba I-digested pGEM-7Zf(+) vector. Nonspecific signals corresponding to 45 and 38 kDa proteins were also shown in both IPTG-induced and noninduced cells. The nonspecific proteins may be products of incomplete translation or proteolytic products of intact PARS.

• Applied Chemistry for Engineering
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• v.24 no.5
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• pp.558-561
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• 2013

In the present work, we constructed a recombinant Escherichia coli with cytoplasmic-expressed phosphate-binding protein (PBP) and investigated its phosphate removal in water phase. When the recombinant bacteria were cultured for 6 h to treat phosphate, the removal efficiencies were 90, 49, and 41% for the treatment of 1.0, 1.5, and 2.0 mg/L phosphate, respectively, indicating good specific phosphate removal of our developed system. Also, cell densities of 2.5 and 5.0 Optical density resulted in high phosphate removal efficiencies and ~80% of 2.0 mg/L phosphate was efficiently removed. A novel biotechnology developed in this study could be effectively employed for resolving eutrophication problem in water body.

• Korean Journal of Plant Resources
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• v.17 no.2
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• pp.161-168
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• 2004

Optimal regeneration conditions of ginseng transformants were studied. It has been known that Ferritin Light Heavy Chain (FLHC) gene remove the several heavy metal by combination, store and transport. To obtain the ginseng tolerant to heavy metal, binary vector was introduced in Agrobacterium by tri-parental mating and then Agrobacterium tumefaciens MP90/FLHC was selected on the AB media and MS media containing kanamycin. Explants were co-cultured with Agrobacterium tumefaciens MP90/FLHC, which contained NPT II as a selectable marker, tadpole ferritin heavy chain (FLHC) gene and human ferritin light chain gene and then a number of embryos were induced. The induced embryo transferred to shooting media consisting of MS medium supplemented with GA 10 mg/L. As a result of examination that induced the normal growth of transfomants, transformants showed the equivalent growth in both root and shoot on the media containing the 1/3 MS.

• Journal of Marine Bioscience and Biotechnology
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• v.1 no.2
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• pp.119-125
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• 2006

To understand the comprehensive mechanisms of biological function for insulin-like growth factor-I (IGF-I) in vertebrates, we have investigated the cDNA sequence of this gene in the korean rockfish (Sebastes schlegeli). The mature form of korean rockfish IGF-I was found to be comprised of 67 amino acid residues, showing about a 7 kDa molecular weight. In this study, we used the polymerase chain reaction (PCR) to obtain a korean rockfish IGF-I (KR IGF-I) cDNA fragment, and methods of rapid amplification of cDNA ends (RACE) to obtain a full length of the KR IGF-I sequence. The KR IGF-I encoded for a predicted amino acid sequence showed identities of 93.6 %, 90.7 %, and 85.4 % in comparison with flounder, chinook salmon, and human IGF-I, respectively. To obtain recombinant biologically active polypeptides, korean rockfish B-C-A-D domains were amplified using the PCR, then the isolated cDNA was expressed in the E. coli BL21(DE3). The recombinant KR IGF-I protein biological function was measured by stimulation of [$^3H$] thymidine incorporation, suggesting the cDNA codes for the korean rockfish proIGF-I.