• Proceedings of the Zoological Society Korea Conference
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• 1998.10b
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• pp.113-113
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• 1998

In Saccharomyces cerevisiae UV irradiation and a variety of chemical DNA -damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of DNA -damage inducible genes is PHRI, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHRI require an upstream activation sequence, UASPHRI. Here we report the identification of the UlvIE6 gene of S. cerevisiae as a regulator of UASPHRl activity. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHRI is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UNIE6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHRI mRNA, and increases the UV sensitivity of a rad2 mutant. The results suggest that UM E6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. Furthermore, the upstream repression sequence, URSPHRI, is required for repression and damage-induced expression of PHRl. Here we show identification of YER169W and YDR096W as putative regulators acting through $URS_{PHRI}$. These open reading frames were designated as RPHI (YERl69W) and RPH2 (YDR096W) indicating regulator of PHRI. Simultaneous disruption of both genes showed a synergistic effect, producing a four-fold increase in basal level expression and a similar decrease m the induction ratio following treatment of methyl methanesulfonate(MMS). Mutation of the sequence ($AG_4$) bound by Rphlp rendered the promoter of PHRI insensitive to changes in RPHI or RPH2 status. The data suggest that RPHI and RPH2 act as damage-responsive negative regulators of PHRI. Surprisingly, the sequence bound by Rphlp in vitro is found to be $AG_4$ which is identical to the consensus binding site for the regulators Msn2p and Msn4p involved in stress-induced expression. Deletion of MSN2 and MSN4 has little effect on the induction$.$ ratio following DNA damage. However, all deletions led to a significant decrease in basal-level and induced expression of PHRI. These results imply that MSN2 and MSN4 are positive regulators of P HRI but are not required for DNA damage repression. [Supported by grant from NIH]om NIH]

• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1460-1468
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• 2007

In this study, an approx. 2.5-kb gene fragment including the catalase gene from Rhodospirillum rubrum S1 was cloned and characterized. The determination of the complete nucleotide sequence revealed that the cloned DNA fragment was organized into three open reading frames, designated as ORF1, catalase, and ORF3 in that order. The catalase gene consisted of 1,455 nucleotides and 484 amino acids, including the initiation and stop codons, and was located 326 bp upstream in the opposite direction of ORF1. The catalase was overproduced in Escherichia coli UM255, a catalase-deficient mutant, and then purified for the biochemical characterization of the enzyme. The purified catalase had an estimated molecular mass of 189 kDa, consisting of four identical subunits of 61 kDa. The enzyme exhibited activity over a broad pH range from pH 5.0 to pH 11.0 and temperature range from $20^{\circ}C$ to $60^{\circ}C$C. The catalase activity was inhibited by 3-amino-1,2,4-triazole, cyanide, azide, and hydroxylamine. The enzyme's $K_m$ value and $V_{max}$ of the catalase for $H_2O_2$ were 21.8 mM and 39,960 U/mg, respectively. Spectrophotometric analysis revealed that the ratio of $A_{406}$ to $A_{280}$ for the catalase was 0.97, indicating the presence of a ferric component. The absorption spectrum of catalase-4 exhibited a Soret band at 406 nm, which is typical of a heme-containing catalase. Treatment of the enzyme with dithionite did not alter the spectral shape and revealed no peroxidase activity. The combined results of the gene sequence and biochemical characterization proved that the catalase cloned from strain S1 in this study was a typical monofunctional catalase, which differed from the other types of catalases found in strain S1.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.111-117
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• 1995

In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

• Korean Journal of Applied Biomechanics
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• v.25 no.4
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• pp.431-440
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• 2015

Objective : The purpose of this study was to investigate the relations between accuracy score and the motions which are performed in PyeongWon Poomsae, thereby developing objectivity in accuracy evaluations for Poomsae competitions. Method : The subjects were six male Poomsae players (age: $21.2{\pm}1.17yrs$, height: $173.4{\pm}3.95cm$, weight: $67.7{\pm}7.65kg$). A three-dimensional motion analysis was performed on the subjects using six high-speed cameras (60 frames/sec) and subjects' performed motions were evaluated by 5 evaluators. The entire Pyeong Poomsae was divided into 13 events and 9 phases; there were three pairs of symmetric phases among them: front kick & turning side kick phases (3PH, 3-1PH), arm motion & stance phases (4PH, 4-1PH), side kick with Hakdari-stance phases (5PH, 5-1PH). Performance time, change and range of COM, height of vertex, and foot of side kicks were analysed. The Data was analysed utilizing correlation analysis. Results : There was a positive correlation between accuracy score and the difference between right and left range of COM (X direction) at 4PH (r=0.921, p=0.009). Conclusion : The results of our study indicate that it is necessary to consider some of objective criterion such as performance time, COM range, and symmetrical movements in accuracy evaluations of Poomsae competitions.

• Korean Journal of Applied Biomechanics
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• v.25 no.1
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• pp.1-10
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• 2015

Objective : The purpose of this study was to find some kinetic variable's relationships between personal records and low records in female high jump. Methods : Collected data of the subjects(N=8, ages: $25.5{\pm}1.85$, height: $173{\pm}5.83$, mass: $54.75{\pm}6.36$ personal record: $1.71{\pm}0.04$, low record: $1.62{\pm}0.03$) were used for the last three strides and take-off phase. Five video cameras set in 30frames/s were used for recording. After digitizing motion, the Direct Linear Transformation(DLT) technique was employed to obtain 3-D position coordinates. The kinematic and kinetic factors of distance, velocity, angle, impulse, jerk variables were calculated. A paired t-test was applied for the difference of variables between personal records and lower records and for correlation with performances and variables. The significance level was accepted at p<.05. Results : There was no relationship between pattern of stride and performance. However, rate of change of velocity was related with cental of mass height(CMH) at peak point(PP). Knee, hip, backward lean, foot plant, approach and take off angle showed no difference between best record and low record. Vertical impulse momentum also showed no difference between performances. Conclusion : According to a t-test result, there were significant differences in CMH at PP and jerk at touch down between best record and low record.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2004.06a
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• pp.278-281
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• 2004

The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.6
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• pp.839-846
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• 1994

Changing concepts in fishery science increasingly are recognizing depletion of traditional stocks, utilization of alternate(non-traditional) species, demand for high quality products, and a total resource utilization approach. Innovative practices are occurring in fisheries processing wherein solid and liquid discharges are no longer treated as 'waste,' but rather as valuable feedstocks for recovery of a variety of value-added ('value enhanced') by-products. Among these are protein hydrolysates, soluble proteins and amino acids, proteolytic enzymes, flavor and flavor extracts, pigments, and biopolymers such as chitosan. Properties and applications of this deacetylated derivative of chitin are noted. Crustacean processing by-products are discussed in terms of their serving as materials for generation of natural flavors and flavor extracts, and products such as fish sauces using contemporary enzymatic techniques. Various food and feed applications of fisheries processing by-products are illustrated with increased usage seen in formulated diets for an expanding aquaculture market. Examples are given of aquaculture becoming increasingly significant in global fisheries resource projections. Critical issues in the international seafood industry Include those of seafood quality, processing quality assurance (HACCP), and recognition of the nutritional and health-related properties of fisheries products. A variety of current seafood processing research is discussed, including that of alternate fish species for surimi manufacture and formulation of value-added seafood products from crawfish and blue crab processing operations. Increasing emphasis is being placed on international aspects of global fisheries and the role of aquaculture in such considerations. Coupled with the need for the aquatic food industry to develop innovative seafood products for the 21st century is that of total resource utilization. Contemporary approaches in seafood processing recognize the need to discard the traditional concept of processing 'waste' and adapt a more realistic, and economically sound, approach of usable by-products for food and feed application. For example, in a period of declining natural fishery resources it is no longer feasible to discard fish frames following fillet removal when a significant amount of residual valuable flesh is present that can be readily recovered and properly utilized in a variety of mince-based formulated seafood products.

• BMB Reports
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• v.38 no.1
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• pp.49-57
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• 2005

Major Royal Jelly Protein cDNAs of Apis cerana (AcMRJP) were cloned and characterized. The open reading frames (ORFs) of the AcMRJP1 and AcMRJP2 genes were 1302 and 1392 nucleotides, encoding 433 and 463 amino acid residues, respectively. The sequence divergences between AcMRJP1 and AcMRJP2 and their corresponding protein families in A. mellifera were 0.0618 and 0.0934 at the nucleotide level and 0.0912 and 0.1438 at the protein level, respectively. Phylogenetic analysis supports the orthologous similarity between these proteins. The deduced amino acids indicated high essential amino acid contents of AcMRJP1 and AcMRJP2 (47.5 and 44.8%, respectively). The genomic organization of both AcMRJP1 and AcMRJP2 was determined. Both the AcMRJP1 (3663 bp) and AcMRJP2 (3963 bp) genes contained six exons and five introns, where all boundaries conformed to the GT/AG rule. AcMRJP1 and AcMRJP2 cDNAs were cloned into pET17b, and both the recombinant (r) AcMRJP1 (47.9 kDa) and rAcMRJP2 (51.7 kDa) were expressed in the insoluble form. Western blot analysis and N-terminal sequencing of the solubilized proteins revealed successful expression of rAcMRJP1 and rAcMRJP2 in vitro. The yields of the purified rAcMRJP1 and rAcMRJP2 were approximately 20 and 8mg protein per liter of the flask culture, respectively.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1189-1196
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• 2005

The cyclohexanol dehydrogenase (ChnA), produced by Rhodococcus sp. TK6, which is capable of growth on cyclohexanol as the sole carbon source, has been previously purified and characterized. However, the current study cloned the complete gene (chnA) for ChnA and its flanking regions using a combination of a polymerase chain reaction (PCR) based on the N-terminal amino acid sequence of the purified ChnA and plaque hybridization from a phage library of Rhodococcus sp. TK6. A sequence analysis of the 5,965-bp DNA fragment revealed five potential open reading frames (ORFs) designated as partial pte (phosphotriesterase), acs (acyl-CoA synthetase), scd (short chain dehydrogenase), stp (sugar transporter), and chnA (cyclohexanol dehydrogenase), respectively. The deduced amino acid sequence of the chnA gene exhibited a similarity of up to $53\%$ with members of the short-chain dehydrogenase/reductase (SDR) family. The chnA gene was expressed using the pET21 a(+) system in Escherichia coli. The activity of the expressed ChnA was then confirmed (13.6 U/mg of protein) and its properties investigated.

• Earthquakes and Structures
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• v.15 no.5
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• pp.529-540
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• 2018

The influences of initial damage paths and aftershock (AS) spectral shape on the assessment of AS collapse fragility are investigated. To do this, a four-story ductile reinforced concrete (RC) frame structure is employed as the study case. The far-field earthquake records recommended by FEMA P695 are used as AS ground motions. The AS incremental dynamic analyses are performed for the damaged structure. To examine the effect of initial damage paths, a total of six kinds of initial damage paths are adopted to simulate different initial damage states of the structure by pushover analysis and dynamic analysis. For the pushover-based initial damage paths, the structure is "pushed" using either uniform or triangle lateral load pattern to a specified damage state quantified by the maximum inter-story drift ratio. Among the dynamic initial damage paths, one single mainshock ground motion or a suite of mainshock ground motions are used in the incremental dynamic analyses to generate a specified initial damage state to the structure. The results show that the structure collapse capacity is reduced as the increase of initial damage, and the initial damage paths show a significant effect on the calculated collapse capacities of the damaged structure (especially at severe damage states). To account for the effect of AS spectral shape, the AS collapse fragility can be adjusted at different target values of ${\varepsilon}$ by using the linear correlation model between the collapse capacity (in term of spectral intensity) and the AS ${\varepsilon}$ values, and coefficients of this linear model is found to be associated with the initial damage states.