• Title/Summary/Keyword: E vector

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Transformation of Carrot (Daucus carota) Cells Using Binary Vector System (Binary Vector System을 이용한 당근 (Daucus carota) 세포의 형질전환)

  • 양덕조;이성택
    • KSBB Journal
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    • v.5 no.3
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    • pp.247-253
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    • 1990
  • These studies were carried out to obtain the transformant from carrot cells by using binary vector pGA472 with NPT II gene to confer kanamycin resistance in the plant cells. The binary vector pGA472 was mobilized from E. coli MC1000 into A. tumefaciens strains isolated in the Korea, C23-1. K29-1, and disarmed Ti-plasmid PC2760, and A28l using a tri-parental mating method with E. coli HB101/pRK2013. Transconjugants, C23-1/pGA472, K29-1/pGA472, PC2 760/pGA472 and A28l/pGA472 were obtaind on the minimum AB media containing tetracycline and kanamycin, were comfirmed to hold the Ti-plasmid and pGA472 binary vector on the 0.7% agarose gel. Transformed carrot calli were initiated on the MS media supplemented with l00$\mu\textrm{g}$/ml kanamycin and 250$\mu\textrm{g}$/ml carbenicillin after co-cultivation of carrot explant and transconjugant Agrobacteria. Selected callus was grown vigouousley for subculture on the medium containing 100$\mu\textrm{g}$/ml kanamycin, thus indication that the selected callus was transformed with NPT II gene.

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Effect of Exercise and Physical Stresses on the Electrocardiogram (운동부하 및 각종 신체조건이 혈압 및 ECG에 미치는 영향 -제2보- (각종 Stress에 의한 심전도 변화))

  • Park, Won-Kyun;Chae, E-Up
    • The Korean Journal of Physiology
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    • v.16 no.2
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    • pp.129-136
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    • 1982
  • We studied this experiment to compare the effects of exercise and other body conditions: i.e., Flack test, cold pressor test and bicycle ergometry on the electrocardiogram. We had sixty healthy college students who were thirty nine men and twenty one women. Their $mean{\pm}SD$ values of physical characteristics were as follows: age; $22.0{\pm}1.4$, weight; men $61.7{\pm}5.6\;kg$, women $46.2{\pm}7.47\;kg$. We observed the changes of P-Q and Q-T interval, R and T amplitude, mean QRS vector, S-T segment deviation, and P and T vector. The result obtained were summarized as follows: P vector was shifted rightward regardless of the type of stress. T vector was shifted var-in each stress but in the bicycle ergometry T vector was shifted leftward. Mean QRS vector was shifted rightward immediately after the bicycle ergometry. Percentage of the occurrence of the depression of S-T segment was 21.7% at the immediately after the submaximal bicycle ergometry in lead II. The elevation of S-T segment was often observed after the mild stresses. Increased amplitude of T wave in the cold pressor test and decreased amplitude of T wave in the bicycle ergometry were observed. In the bicycle ergometry and other stresses, the precise mechanism of S-T segment changes was unexplained but insufficient repolarization in base or apex of the left ventricle due to heart strain was indicated by so called S-T vector analysis.

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Deletion Analysis of Pichia PGK1 Promoter and Construction of an Episomal Vector for Heterologous Protein Expression in P. pastoris (Pichia PGK1프로모터의 분석과 P. pastoris에 있어 외래단백질발현을 위한 Episomal벡터의 제조)

  • Lee, Sung-Jae;Hong, In-Pyo;Baek, Seon-Yeol;Choi, Shin-Geon
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.184-190
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    • 2007
  • Approximately 2.0 kb of the promoter region of the Pichia pastoris phosphoglycerate kinase gene (PGK1) was reduced to a 266 bp fragment and this minimized portion was used for construction of a new episomal constitutive expression vector in P. pastoris. As an approach to developing a constitutive expression vector in P. pastoris, the GAP promoter region of the Pichia expression vector pGAPZB was replaced with sequentially deleted PGK1 promoter fragments fused to a beta-galactosidase gene. When a lacZ gene was used as a reporter gene, PGK1 promoter strength was lower than that of the constitutive GAP promoter but it was higher than TEF1. We report here the development of the pPGKZ-E vector as a new episomal expression vector for heterologous gene expression by removing non-essential regions of the PGK1 promoter. This broadens the choice of episomal expression vectors for controlled constitutive expression in P. pastoris.

Model order reduction for Campbell diagram analysis of shaft-disc-blade system in 3D finite elements

  • Phuor, Ty;Yoon, GilHo
    • Structural Engineering and Mechanics
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    • v.81 no.4
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    • pp.411-428
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    • 2022
  • This paper presents the Campbell diagram analysis of the rotordynamic system using the full order model (FOM) and the reduced order model (ROM) techniques to determine the critical speeds, identify the stability and reduce the computational time. Due to the spin-speed-dependent matrices (e.g., centrifugal stiffening matrix), several model order reduction (MOR) techniques may be considered, such as the modal superposition (MS) method and the Krylov subspace-based MOR techniques (e.g., Ritz vector (RV), quasi-static Ritz vector (QSRV), multifrequency quasi-static Ritz vector (MQSRV), multifrequency/ multi-spin-speed quasi-static Ritz vector (MMQSRV) and the combined Ritz vector & modal superposition (RV+MS) methods). The proposed MMQSRV method in this study is extended from the MQSRV method by incorporating the rotational-speed-dependent stiffness matrices into the Krylov subspace during the MOR process. Thus, the objective of this note is to respond to the question of whether to use the MS method or the Krylov subspace-based MOR technique in establishing the Campbell diagram of the shaft-disc-blade assembly systems in three-dimensional (3D) finite element analysis (FEA). The Campbell diagrams produced by the FOM and various MOR methods are presented and discussed thoroughly by computing the norm of relative errors (ER). It is found that the RV and the MS methods are dominant at low and high rotating speeds, respectively. More precisely, as the spinning velocity becomes large, the calculated ER produced by the RV method is significantly increased; in contrast, the ER produced by the MS method is smaller and more consistent. From a computational point of view, the MORs have substantially reduced the time computing considerably compared to the FOM. Additionally, the verification of the 3D FE rotordynamic model is also provided and found to be in close agreement with the existing solutions.

A New N-time Systolic Array Architecture for the Vector Median Filter (N-time 시스톨릭 어레이 구조를 가지는 벡터 미디언 필터의 하드웨어 아키텍쳐)

  • Yang, Yeong-Yil
    • Journal of the Institute of Convergence Signal Processing
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    • v.8 no.4
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    • pp.293-296
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    • 2007
  • In this paper, we propose the systolic array architecture for the vector median filter. In the color image processing, the vector signal (i.e. the color) consists of three elements, red, green and blue. The vector median filter is very effective to utilize the correlation among red, green and blue elements. The computational complexity of the proposed architecture for computing the vector median of N vector signals is (N+2) clock periods compared to the (3N+1) clock periods in the previous method. In addition to, the input vector signals can be loaded in serial in the proposed architecture. In the previous method, N input vector signals should be loaded to the vector median filter in parallel at the first clock. The proposed architecture is implemented with FPGA.

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Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.211-218
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    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

Characterization of the ${\beta}-Cyclodextrin$ Glucanotransferase Bacillus firmus var. alkalophilus and Its Expression in E. coli

  • Park, Tae-Hyung;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.811-819
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    • 1999
  • The ${\beta}-CGTase$ gene of alkalophilic Bacillus firmus var. alkalophilus was cloned into E. coli using $pZErO^{TM}-2$ as a vector. The cloned gene encoded a total of 710 amino acid residues consisting of 674 amino acids of the matured protein and 36 amino acids of the signal peptide, including 20 amino acids from the lacZ gene in the vector. Although the cloned ${\beta}-CGTase$ gene did not contain the promoter and start codons, it was expressed by the lac promoter and lacZ start codon in the $pZErO^{TM}$ vector. A comparison was made with the amino acid sequence and ten other CGTases from Bacillus sp. Also, ten highly conserved regions, which are important amino acid residues in catalysis of CGTase, were identified. The lac promoter used for expression of the ${\beta}-CGTase$ gene was induced constitutively in recombinant E. coli even without IPTG possibly because of a lack of the lacI gene in both host and vector, repressing the lacZ gene in the lac operon. Its expression was catabolically repressed by glucose, however, its repression was reduced by soluble starch, mainly because of the extremely high increase of the cAMP level. ${\beta}-CGTase$ can be overproduced in the recombinant E. coli by maintaining intracellular cAMP levels mostly through the intermittent feeding of glucose during cultivation.

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Periplasmic Expression of a Recombinant Antibody (MabB9) in Escherichia coli

  • Chang, Hae-Choon;Kwak, Ju-Won
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.299-304
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    • 1997
  • Expression in the periplasm of Escherichia coli of cloned heavy and light chain cDNAs for Fab fragment of a murine monoclonal antibody MabB9 (${\gamma}2b$, K), specific for human plasma apolipoprotein B-100 of LDL, was studied. For the purpose, a vector for two-cistronic expression of the heavy chain cDNA, at the 5' terminus, and light chain cDNA, at the 3' terminus, was constructed using the signal sequences, pelB (for heavy chain) and ompA (for light chain) in a pET vector system. The constructed vector was transformed into E. coli BL21(DE3). The expressed heavy chain (25 kDa) and light chain (23 kDa) of the antibody molecule were detected in total cell extracts as well as in the periplasmic extracts of E. coli.

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A Novel Approach to Improving the Performance of Randomly Perturbed Sensor Arrays (불규칙하게 흔들리는 센서어레이의 성능향상을 위한 새로운 방법)

  • Chang, Byong-Kun
    • The Journal of the Acoustical Society of Korea
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    • v.14 no.1E
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    • pp.65-72
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    • 1995
  • The effects of random errors in array weight and sensor positions on the performance of a Linearly constrained linear sensor array is analyzed in a weight vector space. It is observed that a nonorthogonality exists between an optimum weight vector and the steering vector of an interference direction du e to random errors. A novel approach to improving the nulling performance by compensating for the nonorthogonality is proposed. Computer simulation results are presented.

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Patch load resistance of longitudinally stiffened webs: Modeling via support vector machines

  • Kurtoglu, Ahmet Emin
    • Steel and Composite Structures
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    • v.29 no.3
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    • pp.309-318
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    • 2018
  • Steel girders are the structural members often used for passing long spans. Mostly being subjected to patch loading, or concentrated loading, steel girders are likely to face sudden deformation or damage e.g., web breathing. Horizontal or vertical stiffeners are employed to overcome this phenomenon. This study aims at assessing the feasibility of a machine learning method, namely the support vector machines (SVM) in predicting the patch loading resistance of longitudinally stiffened webs. A database consisting of 162 test data is utilized to develop SVM models and the model with best performance is selected for further inspection. Existing formulations proposed by other researchers are also investigated for comparison. BS5400 and other existing models (model I, model II and model III) appear to yield underestimated predictions with a large scatter; i.e., mean experimental-to-predicted ratios of 1.517, 1.092, 1.155 and 1.256, respectively; whereas the selected SVM model has high prediction accuracy with significantly less scatter. Robust nature and accurate predictions of SVM confirms its feasibility of potential use in solving complex engineering problems.