• Title/Summary/Keyword: 유전자 예측

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Development of L-Threonine Producing Recombinant Escherichia coli using Metabolic Control Analysis (대사 조절 분석 기법을 이용한 L-Threonine 생산 재조합 대장균 개발)

  • Choi, Jong-Il;Park, Young-Hoon;Yang, Young-Lyeol
    • KSBB Journal
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    • v.22 no.1
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    • pp.62-65
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    • 2007
  • New strain development strategy using kinetic models and metabolic control analysis was investigated. In this study, previously reported mathematical models describing the enzyme kinetics of intracellular threonine synthesis were modified for mutant threonine producer Escherichia coli TF5015. Using the modified models, metabolic control analysis was carried out to identify the rate limiting step by evaluating the flux control coefficient on the overall threonine synthesis flux exerted by individual enzymatic reactions. The result suggested the production of threonine could be enhanced most efficiently by increasing aspartate semialdehyde dehydrogenase (asd) activity of this strain. Amplification of asd gene in recombinant strain TF5015 (pCL-$P_{aroF}$-asd) increased the threonine production up to 23%, which is much higher than 14% obtained by amplifying aspartate kinse (thrA), other gene in threonine biosynthesis pathway.

Characterization of a fad3 cDNA Encoding Microsomal Fatty Acid Desaturase from Arabidopsis thaliana (Arabidopsis thaliana로부터 지방산 불포화효소 유전자의 분석)

  • 박희성;임경준
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.93-97
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    • 1997
  • For the molecular genetic study of cold tolerance mechanism in plants, a cDNA encoding fatty acid desaturase (fad3), converting linoleic acid (18:2, $\omega$-6) to linolenic acid (18:3, $\omega$-3), was isolated from $\lambda$ZAPII Arabidopsis thaliana cDNA expression library by plaque hybridization using fad3 cDNA probe derived from Brassica napus. A 1.8 kb-EcoRI fragment from a lambda clone showing a strong positive hybridization signal was subcloned into pGEM7 and analyzed for its nucleotide sequence. From deduced amino acid sequences, the fad3 gene was revealed to have an open reading frame(ORF) consisting of 386 amino acids with a molecular mass of 44,075 Da. The fad3 gene was compared to chloroplast $\omega$-3 fatty acid desaturase (fad7) and endoplasmic reticulum Δ12 fatty acid desaturase (fad2) to show 70% and 58% amino acid sequence homology, respectively, Especially, amino acids of internal (82 to 151) and carboxy terminal (276 to 333) regions were highly conserved, implying their requisite role for enzymatic functioning of fatty acid desaturases. IPTG-induced fad3 cDNA expression in E. coli cells was suggested to be toxic to bacterial growth.

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Expression Pattern of Acetyl Xylan Esterase of Streptomyces coelicolor A3(2) in Escherichia coli (Escherichia coli에서의 Streptomyces coelicolor A3(2)의 acetyl xylan esterase 발현 양상)

  • 이인숙;윤석원;정상운;오충훈;김재헌
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.83-88
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    • 2003
  • We cloned a gene encoding acetyl xylan esterase(axeA) of Streptomyces coelicolor A3(2) and studied its expression pattern in Escherichia coli. The full sequence of axeA was amplified by PCR. Sequence analysis of the PCR product revealed an open reading frame of 1,008 nucleotides encoding a protein consisted of 335 amino acid residues, with a calculated molecular mass of about 38 kDa. The base sequence showed 98% homology to the same gene of Streptomyces lividans. Two different kinds of acetyl xylan esterases were produced in Escherichia coli(pLacI) by IPTG induction; their molecular weights were 38 kDa and 34 kDa, respectively. Of these, 38 kDa protein seemed to be a total protein holding N-terminal signal peptide region, whereas 34 kDa protein seemed to be a matured protein without signal peptide which was produced by peptide bond cleavage between two amino acid residues of alanine 41 and alanine 42.

Development of a Bi-objective Cycle-free Signal Timing Model Using Genetic Algorithm (유전자 알고리즘을 이용한 이중목적 주기변동 신호시간 결정 모형 개발)

  • 최완석;이영인
    • Journal of Korean Society of Transportation
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    • v.20 no.5
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    • pp.81-98
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    • 2002
  • This paper documents the development of a bi-objective(minimizing delays and Queue lengths) cycle-free signal timing length model using Genetic Algorithm. The model was embodied using MATLAB. the language of technical computing. A special feature of this model is its ability to concurrently manage delays and queue lengths of turning movement concurrently. The model produces a cycle-free signal timing(cycles and green times) for each intersection on the cycle basis. Appropriate offsets could be also accomplished by applying cycle-free based signal timings for respective intersections. The model was applied to an example network which consists of four intersections. The result shows that the model produces superior signal timings to the existing signal timing model in terms of managing delays and queue lengths of turning movements.

Selection of next-generation antigen protein for diagnosis of pfhrp2/pfhrp3 gene deleted plasmodium falciparum based on bioinformatics (pfhrp2/pfhrp3 유전자 결여 열대열 말라리아 특이 진단을 위한 생물정보학 기반 차세대 항원 단백질 선정)

  • Seo, Seung Hwan;Lee, Jihoo;Choi, Jae-Won;Kim, Hak Yong
    • Proceedings of the Korea Contents Association Conference
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    • 2016.05a
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    • pp.187-188
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    • 2016
  • 열대열 말라리아(Plasmodium falciparum, P. falciparum, P. f) 신속진단키트의 경우, P. falciparum에 특이적인 단백질로써 Histidine Rich Protein 2 (PfHRP2)가 사용되고 있다. 그러나 최근 연구에서 남아메리카와 중앙아메리카를 중심으로 pfhrp2/pfhrp3 유전자가 결여된 P. falciparum 열원충이 나타나는 것으로 보고된 바 있다. 본 연구에서는 생물정보학을 기반으로 PfHRP2 항원 단백질을 대체할 수 있는 새로운 P. falciparum 특이 항원 단백질을 선정하고자, PlasmoDB에서 5,777개의 P. falciparum 관련 단백질 리스트를 얻었다. 이후 NCBI BLAST를 통해 단백질 아미노산 서열을 분석하고 정상인에게 존재하지 않으며, 동시에 다른 말라리아 열원충(P. vivax, P. ovale, P. malariae, P. knowlesi)에도 존재하지 않는 P. falciparum 특이 아미노산 서열을 가진 단백질 15개를 추출하였다. IEDB analysis를 이용하여 에피토프, 수용성, 베타-턴, 접근성, 유연성, 면역원성을 분석하여 높은 평균값을 갖는 상위 3개 단백질을 선별하였다. KEGG pathway와 EMBL-EBI를 통해 선별된 3개 단백질의 혈액내 검출 가능성 및 아미노산 서열의 보존성을 분석하여 최종적으로 Glutamate-Rich Protein (GLURP)을 선정하였다. AIDA를 통해 단백질 아미노산 서열을 이용한 3차 구조 예측으로 GLURP의 구조 및 항체와의 결합을 도식화하였다. 최종적으로 선정한 GLURP는 pfhrp2/pfhrp3 유전자 결여 P. falciparum까지 특이적으로 진단이 가능하여 차세대 P. falciparum 특이 신속진단키트 개발에 도움이 될 수 있을 것으로 기대한다.

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Task Sequence Optimization for 6-DOF Manipulator in Press Forming Process (프레스 공정에서 6자유도 로봇의 작업 시퀀스 최적화)

  • Yoon, Hyun Joong;Chung, Seong Youb
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.2
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    • pp.704-710
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    • 2017
  • Our research team is developing a 6-DOF manipulator that is adequate for the narrow workspace of press forming processes. This paper addresses the task sequence optimization methods for the manipulator to minimize the task-finishing time. First, a kinematic model of the manipulator is presented, and the anticipated times for moving among the task locations are computed. Then, a mathematical model of the task sequence optimization problem is presented, followed by a comparison of three meta-heuristic methods to solve the optimization problem: an ant colony system, simulated annealing, and a genetic algorithm. The simulation shows that the genetic algorithm is robust to the parameter settings and has the best performance in both minimizing the task-finishing time and the computing time compared to the other methods. Finally, the algorithms were implemented and validated through a simulation using Mathworks' Matlab and Coppelia Robotics' V-REP (virtual robot experimentation platform).

Evaluation of the preservation state of human skeletal remains using real-time PCR (출토 인골 DNA의 real-time PCR 정량에 의한 보존상태 평가 연구 - 부여 오수리 출토 인골을 중심으로 -)

  • Kwon, Eun-Sil;Cho, Eun-Min;Kim, Sue-Hoon;Kang, Soyeong
    • 보존과학연구
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    • s.32
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    • pp.171-183
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    • 2011
  • In this study molecular genetic analysis was carried out on 4 human skeletal remains from Osuri, Buyeo. We showed that real-time PCR is the method of the choice to assess the initial number of genuine ancient DNA molecules. Human mitochondrial DNA quantification was accomplished by the real-time PCR for the cytochrome b gene of the mitochondria. Histological results proved to be a good potentiality for biochemical analysis using biomolecule. The level of specimen's preservation state was proved that level of quantitative result was BO-04, BO-01, BO-03, BO-02. Continually, we showed that biochemical and biomolecule results for the level of preservation state were similar. This study will be useful to important material for predicting biochemistry and biology analysis of the ancient bone.

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A Study on the Training Optimization Using Genetic Algorithm -In case of Statistical Classification considering Normal Distribution- (유전자 알고리즘을 이용한 트레이닝 최적화 기법 연구 - 정규분포를 고려한 통계적 영상분류의 경우 -)

  • 어양담;조봉환;이용웅;김용일
    • Korean Journal of Remote Sensing
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    • v.15 no.3
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    • pp.195-208
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    • 1999
  • In the classification of satellite images, the representative of training of classes is very important factor that affects the classification accuracy. Hence, in order to improve the classification accuracy, it is required to optimize pre-classification stage which determines classification parameters rather than to develop classifiers alone. In this study, the normality of training are calculated at the preclassification stage using SPOT XS and LANDSAT TM. A correlation coefficient of multivariate Q-Q plot with 5% significance level and a variance of initial training are considered as an object function of genetic algorithm in the training normalization process. As a result of normalization of training using the genetic algorithm, it was proved that, for the study area, the mean and variance of each class shifted to the population, and the result showed the possibility of prediction of the distribution of each class.

Genetic Polymorphism of Epoxide Hydrolase and GSTM1 in Chronic Obstructive Pulmonary Disease (만성폐쇄성폐질환 발생에 Epoxide hydrolase와 GSTM1유전자 다형성의 의의)

  • Park, Sang Sun;Kim, Eun Joung;Son, Chang Young;Wi, Jeong Ook;Park, Kyung Hwa;Cho, Gye Jung;Ju, Jin Young;Kim, Kyu Sik;Kim, Yu II;Lim, Sung Chul;Kim, Young Chul;Park, Kyung Ok;Na, Kook Joo
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.1
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    • pp.88-97
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    • 2003
  • Background : Although smoking is a major cause of chronic obstructive pulmonary disease (COPD), only 10-20% of cigarette smokers develop symptomatic COPD, which suggests the presence of genetic susceptibility. This genetic susceptibility to COPD might depend on variations in the activities of the enzyme that detoxify hazardous chemical products, such as microsomal epoxide hydrolase (mEPHX) and glutathione-S transferase M1 subunit (GSTM1) genes. Methods : The genotypes of 58 patients with COPD, and 79 age matched control subjects, were determined by a polymerase chain reaction, followed by restriction fragment length polymorphism (PCR-RFLP) for the mEPHX, and multiplex PCR for the GSTM1. Results : GSTM1 was deleted in 53.3% of the subjects. There was no difference in GSTM1 deletion rates between the COPD patients (32/58, 55.2%) and the control subjects (41/79, 51.9%). The combination patterns of two polymorphisms of mEPHX showed slow enzyme activity in 29(21.2%), normal in 73(53.3%) and fast in 32(23.4%). The COPD group (7/57, 12.3%) showed a significantly lower incidence of slow enzyme activity compared to the control subjects (22/77, 28.6%, p<0.05). However, when the COPD and control groups were compared with smokers only, there were no significant differences in the genotypes of GSTM1 and mEPHX. Conclusion : The genotypes of GSTM1 and mEPHX were not significant risk factors of COPD in this cohort of study.

Genetic Diversity of the Slender Shinner(Pseudopuntungia tenuicorpa) and Its Conservational Implications (가는돌고기(Pseudopuntungia tenuicorpa) 보전을 위한 유전적 다양성 연구)

  • Kim, Dong-Young;Suk, Ho Young
    • Korean Journal of Ichthyology
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    • v.32 no.2
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    • pp.39-48
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    • 2020
  • The slender shinner (Pseudopungtungia tenuicorpa), a tiny freshwater fish of about 8 to 10 cm belonging to Cyprinidae, is an endangered species found only in the Han and Imjin Rivers on the Korean Peninsula. During the breeding season, this species spawns in nests of Coreoperca herzi, a predator of this species, or small crevices on rocks. This unique reproductive ecology can make this species more vulnerable to anthropogenic perturbance that can further limit the places to spawn. Here, mtDNA and microsatellite loci were analyzed to identify the genetic diversity and structure of slender shinners and further to provide the basic data necessary for the conservation planning of this species. A total of 28 polymorphic microsatellite markers were developed using Illumina paired-end sequencing, and 67 slender shinners collected from three localities in the Han River were genotyped using these loci. This species showed a remarkably high level of genetic diversity with mean expected heterozygosity of 0.914 and mean allele number per locus of 27.9, and no signature of drastic demographic decline was detected. As a result of our microsatellite analysis, the genetic structure between the two stems of the Han River, North Han and South Han, was prominent. Such a genetic structure was also evident in the sequence analysis of 14 haplotypes obtained from mtDNA control region. Although slender shinners are only found in very limited areas around the world, the genetic structure indicates that there is a block of gene flow among the populations, which should be reviewed in the future if management and restoration of this species is needed.