• 생명과학회지
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• 제18권11호
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• pp.1592-1599
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• 2008

$\beta$-Lactam계 항생물질에 강한 내성을 가지는 균주 Bacillus sp. J105가 생산하는 $\beta$-lactamase의 유전자를 E. coli DH5$\alpha$에 cloning하였다. Cosmid vector pLAFR3을 이용하여, Sau3AI 으로 부분 분해한 chromosomal DNA와 BamHI으로 처리한 pLAFR3을 ligation하였다. In vitro packaging kit를 사용하여 E. coli에 형질도입 하였으며 $\beta$-lactamase양성 clone주를 획득하였다. 이 recombinant plasmid ($\beta$-lac+)를 pACYC184 (4.2kb) vector를 사용하여 subcloning 하여 최종 $\beta$-lactamase의 활성이 있는 6.4 kb 단편이 포함된 pKL11${\Delta}4.6$을 제작하였다. 이 단편을 DNA 염기서열을 분석한 결과 309개의 아미노산으로 구성된 $\beta$-lactamase를 코딩하는 927 bp를 포함하고 있었다. 클로닝된 $\beta$-lactamase 유전자의 upstream을 포함하는 170 bp의 염기서열을 분석한 결과, B. thuringinesis와 B. cereus 유래의 $\beta$-lactamase 유전자의 upstream 부위와 97%의 일치를 보였다. 본 연구에서 클로닝된 $\beta$-lactamase의 아미노산을 서열을 NCBI BLAST program을 이용하여 분석해 본 결과 B. thuringinesis와 B. cereus의 $\beta$-lactamase와 각각 97%와 94%의 일치를 보였다. 또한 계통도 분석 결과 역시 본 연구에서 클로닝된 $\beta$-lactamase의 아미노산을 서열은 B. thuringinesis와 B. cereus 와 유전학적으로 아주 밀접한 관계를 보여주었다. 이 pKL11-${\Delta}4.6$를 E. coli에서 형질전환 시켜 발현 양상을 조사해 본 결과 $\beta$-lactamase의 secretion efficiency는 약 $4{\sim}5%$％였다. E. coli의 세포 내 단백질로부터 $\beta$-lactamase를 정제하여 분자량을 확인한 결과 31 kDa로 wild type의 분자량과 일치함을 확인하였다.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.178-186
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• 1995

Streptomyces albus wild type ATCC 21838 produced salinomycin, polyether antibiotic. To clone genes related salinomycin production, a genomic library was screened using actI as a DNA hybridization probe. pWHM 210 was isolated, which contained an approximately 24 kb of insert DNA. A 3.8 kb region in the 24 kb insert DNA was hybridized to actI and the nucleotide sequence of this region was determinied. Two open reading frames found in the same direction were homologous to genes for $\beta$-keto acyl synthase/acyl transferase and chain length determining factor in type II PKS (polyketide synthase). The genes were components of minimal type II PKS genes, highly conserved and showed the strong simiarity to other type II PKS genes known today.

• Journal of Interdisciplinary Genomics
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• 제1권2호
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• pp.15-18
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• 2019

Gaucher disease (GD) is an autosomal recessive inborn error of metabolism resulting from a deficiency in ${\beta}$-glucocerebrosidase (GBA) activity that leads to the accumulation of glucocerebroside in macrophages in multiple organs, such as the bone marrow, liver, spleen, and brain. GD can be classified into three clinical types: type 1 (non-neuropathic form, OMIM #230800); type II (acute neuropathic form, OMIM #230900); and type III (chronic neuropathic form, OMIM #231000). Type III is the subacute form of neuropathic GD. The best available treatment for GD is long-term enzyme (imiglucerase) replacement therapy (ERT) performed every two weeks. This report describes the long-term clinical course of a patient with type III GD who was treated with ERT for 18 years.

• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2005년도 BIOINFO 2005
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• pp.325-330
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• 2005

We are generally interested in the analysis, detection and prediction of structural motifs in proteins, in order to infer compatibility of amino acid sequence to structure in proteins of known three-dimensional structure available in the Protein Data Bank. In this context, we are analyzing some of the well-characterized structural motifs in proteins. We have analyzed simple structural motifs, such as, ${\beta}$-turns and ${\gamma}$-turns by evaluating the statistically significant type-dependent amino acid positional preferences in enlarged representative protein datasets and revised the amino acid preferences. In doing so, we identified a number of ‘unexpected’ isolated ${\beta}$-turns with a proline amino acid residue at the (i+2) position. We extended our study to the identification of multiple turns, continuous turns and to peptides that correspond to the combinations of individual ${\beta}$ and ${\gamma}$-turns in proteins and examined the hydrogen-bond interactions likely to stabilize these peptides. This led us to develop a database of structural motifs in proteins (DSMP) that would primarily allow us to make queries based on the various fields in the database for some well-characterized structural motifs, such as, helices, ${\beta}$-strands, turns, ${\beta}$-hairpins, ${\beta}$-${\alpha}$-${\beta}$, ${\psi}$-loops, ${\beta}$-sheets, disulphide bridges. We have recently implemented this information for all entries in the current PDB in a relational database called ODSMP using Oracle9i that is easy to update and maintain and added few additional structural motifs. We have also developed another relational database corresponding to amino acid sequences and their associated secondary structure for representative proteins in the PDB called PSSARD. This database allows flexible queries to be made on the compatibility of amino acid sequences in the PDB to ‘user-defined’ super-secondary structure conformation and vice-versa. Currently, we have extended this database to include nearly 23,000 protein crystal structures available in the PDB. Further, we have analyzed the ‘structural plasticity’ associated with the ${\beta}$-propeller structural motif We have developed a method to automatically detect ${\beta}$-propellers from the PDB codes. We evaluated the accuracy and consistency of predicting ${\beta}$ and ${\gamma}$-turns in proteins using the residue-coupled model. I will discuss results of our work and describe databases and software applications that have been developed.

• Composites Research
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• 제13권2호
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• pp.30-39
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• 2000

고급복합재료(advanced composite material)는 여러 가지 훌륭한 장점에도 불구하고 건설기술자에게는 그 이론이 너무 복잡하여 건설구조에 대한 응용이 지연되고 있다. [$\alpha$/$\beta$/$\beta$/$\gamma$/$\alpha$/$\alpha$/$\beta$]r, $\alpha$=-$\beta$, $\gamma$=$0^{\circ}$또는 $90^{\circ}$와 같은 몇 가지 배향각을 갖은 적층판은, r이 증가함에 따라, $B_{i,j}$ 강성이 0으로 접근한다(1995). 또한 건설구조물은 그 치수가 크고 r이 증가해도 지간 대 두께비가 충분히 커서, 횡방향 변형율($varepsilon_{xz}$, $varepsilon_{yz}$)의 영향이 무시될 수 있다. 이 경우 판의 지배방정식은 간단한 특별직교 이방성 판의 지배방정식으로 변화된다. 김덕현은 토목구조물에 흔히 적용될 수 있는 이러한 배향각을 가진 적층판의 지배방정식을 취급이 간단한 특별직교 이방성 판의 경우로 변경시킨 다음 해석을 더 간단히 진행할 수 있도록 기술자들이 유사 등방성 상수(quasi-isotropic constant)를 사용하여 낯익은 등방성판의 방정식으로 답을 구하고 그가 개발한 "수정계수(correction factor)"를 사용하여 "정확한"해를 구하는 방법을 발표한 바 있다(1991).이 논문에는 [$\alpha$/$\beta$/$\beta$/$\gamma$/$\alpha$/$\alpha$/$\beta$]r 배향각을 갖는 적층판을, 일반기술자들이 이해하기 쉬운 본 논문의 방법으로 해석하는 방법을 제시하고 기술자들의 참고자료로 사용될 수 있는 여러 가지 경우에 대한 고유진동수의 계산결과가 제공되고 있다. 고유진동수의 계산결과가 제공되고 있다.

• 대한약침학회지
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• 제19권2호
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• pp.163-166
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• 2016

Objectives: Quercetin is a flavonoid and an important dietary constituent of fruits and vegetables. In recent years, several pharmacological activities of quercetin, such as its neuroprotective activity and, more specifically, its anti-convulsant effects in animal models of epilepsy, have been reported. This study evaluated the role of quercetin pretreatment on gene expression of ${\gamma}$-amino butyric acid type A ($GABA_A$) receptor beta subunits in kainic acid (KA)-induced seizures in mice. Methods: The animals were divided into four groups: one saline group, one group in which seizures were induced by using KA (10 mg/kg) without quercetin pretreatment and two groups pretreated with quercetin (50 and 100 mg/kg) prior to seizures being induced by using KA. Next, the messenger ribonucleic acid (mRNA) levels of the $GABA_A$ receptor ${\beta}$ subunits in the hippocampus of each animal were assessed at 2 hours and 7 days after KA administration. Quantitative real-time polymerase chain reaction (RT-PCR) assay was used to detect mRNA content in hippocampal tissues. Results: Pretreatments with quercetin at doses of 50 and 100 mg/kg prevented significant increases in the mRNA levels of the ${\beta}_1$ and the ${\beta}_3$ subunits of the $GABA_A$ receptor at 2 hours after KA injection. Pretreatment with quercetin (100 mg/kg) significantly inhibited ${\beta}_1$ and ${\beta}_3$ gene expression in the hippocampus at 7 days after KA injection. But, this inhibitory effect of quercetin at 50 mg/kg on the mRNA levels of the ${\beta}_3$ subunit of the $GABA_A$ receptor was not observed at 7 days after KA administration. Conclusion: These results suggest that quercetin (100 mg/kg) modulates the expression of the $GABA_A$ receptor ${\beta}_1$ and ${\beta}_3$ subunits in the KA model of epilepsy, most likely to prevent compensatory responses. This may be related to the narrow therapeutic dose range for the anticonvulsant activities of quercetin.

• 대한유전성대사질환학회지
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• 제5권1호
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• pp.65-75
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• 2005

목적: 뮤코지방증 유형 II와 III은 리소좀 효소인N-acetylglucosaminyl-1-phosphotransferase (UDP-N-acetylglucosamine, GlcNAc-phospho-transferase)의 결손에 의해 초래되며, 상염색체 열성으로 유전되는 질환이다. ${\alpha}/{\beta}/{\gamma}$ subunit로 구성되는 이 효소의 결핍으로 인해 리소좀으로 운반되는 수십 종류의 효소들에 mannose-6-phosphate(M6P)를 부착하는 과정에 장애가 생겨 분해되지 않은 물질이 축적되어 질병이 초래된다. 이 질환에 있어서 산전 진단과 유전 상담을 위해서는 상기 효소의 해당 유전자가 밝혀져 있어야 하나, 현재 이 효소의 ${\gamma}$ subunit를 암호화하는 유전자가 GNPTAG에 해당된다고 밝혀져 있을 뿐 ${\alpha}/{\beta}$ subunit을 암호화하는 GNPTA에 해당하는 유전자는 밝혀져 있지 않고 돌연변이 역시 보고된 적이 없다. 본 연구는 리소좀 효소의 인산화에 관여하는 N-acetylglucosamine-1-phospho-transferase의 결함이 있는 환자의 섬유아세포를 이용하여 리소좀 연관 신규 유전자를 찾아내고, 그 유전자를 대상으로 돌연변이를 규명하고자 하였다. 방법: 이를 위해 5명의 환자와5명의 연령, 성별이 일치하는 정상아의 섬유아세포를 계대 배양하여 이 세포를 이용하여 수행한 subtractive hybridization을 통해 신규 유전자를 탐색하고, 신규 유전자를 대상으로 돌연변이 분석을 수행하였다. 결과: 연구 결과 환자에서 발현이 증가된 유전자 73개와 발현이 감소된 유전자 50개를 밝혀냈다. 분석된 유전자 중에서 MGC4170이환자에서는 발현되지 않으나 정상인에서는 발현됨을 발견하였고, 이 유전자가 아직 밝혀지지 않은 GNPTA로 확인되어 환자를 대상으로 돌연변이 분석을 시행하였다.분석 결과 기존에 돌연변이가 보고되었던 GNPTAG에는 돌연변이가 없었으나, MGC4170에는 7개의 돌연변이가 발견되었다. 본 연구는 GlcNAc-phosphotransferase의 ${\alpha}/{\beta}$ subunit에 해당되는 MGC4170의 최초 돌연변이 보고이다. 결론: 본 연구를 통해 뮤코지방증 II형과 III형에서 발현되는 유전자 군을 파악할 수 있었으며, 동시에 MGC4170 돌연변이를 규명함으로써 이 질환의 병리 기전 연구와 산전 진단에 기여하고자 한다.

• 한국작물학회지
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• 제39권1호
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• pp.63-72
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• 1994

보리의 이용성을 검토하기 위하여, 찰성 부정형 Shrunken endosperm 유전자를 결합하여 육성한 Isogenic line을 이용하여, 취반 및 제분에 대한 제특성을 조사한 바 그 결과를 요약하면 다음과 같다. 1. 보리의 배유 전분 특성이 다른 Isogenic type 간에 정곡 천립중, 회분, 단백질 함량은 별차이가 없었으나 Amylose함량, $\beta$-glucan viscosity와 취반특성은 뚜렷한 차이를 보였다. 2. 취반특성중 흡수율과 퍼짐성은 천립중, Amylose 함량과 부의 상관이 있었고, 용출고형물량과 점도간에도 부의 상관이 있었다. 3, 흡수율과 퍼짐성은 배유 전분 특성과 입의 크기에 의해 영향을 받았으며 그 결과 찰성 Isogenic type이 높았고 부정형 전분 Isogenic type이 가장 낮았으며 종실의 크기가 작은 것이 큰 것에 비해 높았다. 4. 보리밥의 Mixogram pattern은 전분 특성에 따라 차이가 있었으며 부정형 전분이 가장 많이 으깨어졌다. 5. 제분 특성은 메성이면서 부정형인 Isogenic type이 좋았고 찰성의 Shrunken endosperm Isogenic type은 대단히 불량하였다.

• The Korean Journal of Physiology and Pharmacology
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• 제22권5호
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• pp.493-501
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• 2018

The present study was carried out with the hypothesis that combination of canagliflozin and omega-3 fatty acid may have potential effect on insulin level, insulin resistance, cardiac biomarkers, inflammatory cytokines and histological studies in type 2 diabetes mellitus (DM). Type 2 DM was induced by injecting nicotinamide (120 mg/kg, i.p.) 15 min before STZ (60 mg/kg) injection. Canagliflozin (5 and 10 mg/kg) and omega-3 fatty acid (300 mg/kg) were given for 28 days after confirmation of diabetes. Biochemical estimations revealed elevated levels of glucose, insulin, HOMA-R and inflammatory cytokines in diabetic group. Daily dosing of alone canagliflozin and omega-3 fatty acid slightly reduced elevated levels of glucose, insulin, HOMA-R and inflammatory cytokines ($IL-1{\beta}$, IL-2, and $TNF{\alpha}$), whereas canagliflozin and omega-3 fatty acid combination has reduced these biochemical parameters significantly when compared with diabetic group. Similarly in diabetic group the levels of cardiac biomarkers such as lipid profile, LDH, CKMB and troponin were significantly increased. Elevated levels of cardiac biomarkers were significantly reduced after daily dosing of alone canagliflozin and omega-3 fatty acid. Canagliflozin and omega-3 fatty acid combination has offered better improvement in cardiac biomarkers compared to alone canagliflozin and omega-3 fatty acid. Histopathological analysis also supported the above hypothesis that combination therapy (canagliflozin and omega-3 fatty acid) offered better protection against degenerative changes in ${\beta}-cells$ of pancreas as compared to alone treatment with these drugs. Thus the present study revealed that canagliflozin and omega-3 fatty acid can be used as potential combination therapy in type 2 DM along with cardiac complication.

• 한국학교보건학회지
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• 제32권2호
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• pp.67-76
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• 2019

Purpose: The purpose of this study was to explore self-directed learning ability and its affecting factors among undergraduate students. Methods: A descriptive cross-sectional study design was used. Using a convenience sampling method, data were collected from 196 undergraduate students enrolled in one national university. Negative psychological tendency (i.e. Type D personality), academic engagement, and self-directed learning ability were assessed using a structured questionnaire. Data were analyzed by descriptive statistics, t-test, and analysis of covariance, Pearson's correlation coefficients, and stepwise multiple linear regression, using SPSS/WIN 23.0 program. Results: The mean age of the students was $21.61{\pm}2.40years$ and 56.6% were male students. Approximately, one third (n=67, 34.2%) of the students had Type D personality. The average scores of academic engagement and self-directed learning ability were $3.01{\pm}1.14$ and $3.46{\pm}0.50$, respectively. After controlling for sociodemographic variables, the Type D personality and academic engagement were significant predictors of self-directed learning ability (${\beta}=.64$, p<.001; ${\beta}=-.13$, p=.021, respectively). This model explained 53.6% of the variance in self-directed learning ability. Conclusion: The study identified that Type D personality and academic engagement affect self-directed learning ability of undergraduate students, one in a negative way, the other in a positive way. Educators and educational policy makers need to make efforts to include interventions and strategies that increase academic engagement and change negative psychological dispositions such as D-type personality in the undergraduate education curriculum.