• Journal of Life Science
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• v.10 no.4
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• pp.422-430
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• 2000

In order to search for the effects of Shine-Dalgarno (SD) sequence and nucleotide sequence of spacer region (SD-ATG) on bGH expression, oligonucleotides containing random SD sequences and a spacer region were chemically synthesized. The distance between SD region and initiation codon (ATG) was fixed to 9 nucleotides in length. The expression vectors have been constructed using pT7-1 vector containing a T7 promoter. Positive clones were screened with colony hybridization and named pT7A or pT7B plasmid series. The selected clones were confirmed by DNA sequencing and finally, 19 clones having various SD combinations were obtained. When bovine growth hormone was induced by IPTG in E. coli BL21(DE3), all cells harboring these plasmids produced a detectable level of bGH in western blot analysis. However, various SD sequences did not affect on bGH expression, indicating that the sequences of SD and the spacer region did not sufficiently destabilize mRNA secondary structure of bGH gene. Therefore, these results indicate that the disruption of mRNA secondary structure might be a major factor for regulating bGH expression in the translational initiation process.

• Journal of Aquaculture
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• v.13 no.1
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• pp.29-37
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• 2000

In previously study we isolated several fish matrix attachment regions (MARs) capable of replicating the plasmid by itself. In this study we construct a fish expression vector pBaEGFP(+) containing mud loach ${\beta}$-actin promoter EGFP as reporter gene and SV40 signal. To analyze the effects of the fish expression vector respectively. The fish ARS containing constructs pBaEGFP(+)-ARSs were transfected cells with pBaEGFP(+)-ARS101 and pBaEGFP(+)-ARS223 reduced 10 days to 25 days and then was constant to 30 days after transfection while that of the control vector without ARS element was basal level. The intensity of both constructs showed about 30fold of the intensity compared with the control vector on 30days after transfection individually .E. coli back-transformation analysis shows that pBaEGFP(+)-ARS223 and pBaEGFP(+)-ARS905 maintain in episomal state at least 30 days after transfection. The result indicates that both may be able to replicate the vector in BF-2 cell. Therefore the matrix-attached ARSs enhancing expression of the reporter gene might be useful as a component o the expression vector for transgenic studies.

• KIPS Transactions on Software and Data Engineering
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• v.3 no.3
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• pp.135-138
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• 2014

This paper devices a facial expression recognition method robust to overfitting using 2D-DCT and EHMM algorithm. In particular, this paper achieves enhanced recognition performance by setting up a large window size for 2D-DCT feature extraction and extracting the observation vectors of EHMM. The experimental results on the CK facial expression database and the JAFFE facial expression database showed that the facial expression recognition accuracy was improved according as window size is large. Also, the proposed method revealed the recognition accuracy of 87.79% and showed enhanced recognition performance ranging from 46.01% to 50.05% in comparison to previous approaches based on histogram feature, when CK database is employed for training and JAFFE database is used to test the recognition accuracy.

• The Plant Pathology Journal
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• v.33 no.4
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• pp.429-433
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• 2017

Chrysanthemums (Chrysanthemum morifolium) are susceptible to tobacco mosaic virus (TMV). TMV-based expression vectors have been used in high-throughput experiments for production of foreign protein in plants and also expressing green fluorescent protein (GFP) to allow visualization of TMV movement. Here, we used TMV expressing the GFP to examine the infection of chrysanthemum by a TMV-based expression vector. Viral replication, movement and GFP expression by TMV-GFP were verified in upper leaves of chrysanthemums up to 73 days post inoculation (dpi) by RT-PCR. Neither wild-type TMV nor TMV-GFP induced symptoms. GFP fluorescence was seen in the larger veins of the inoculated leaf, in the stem above the inoculation site and in petioles of upper leaves, although there was no consistent detection of GFP fluorescence in the lamina of upper leaves under UV. Thus, a TMV-based expression vector can infect chrysanthemum and can be used for the in vivo study of gene functions.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.381-391
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• 1999

During the last 20 years, transgenic animal technology has provided revolutionary new opportunities in many aspects of agriculture and biotechnology. Several gene delivery systems including pronuclear injection, retroviral vectors, sperm vectors, and somatic cell cloning have developed for making transgenic animals. In the future major improvements in transgenic animal generation will be mainly covered by somatic cell cloning technology. Many factors affecting integration frequency and expression of the transgenes should be overcome to facilitate the industrial applications of transgenic technology. Transgenic animal technology has settled down in some areas of the biotechnology, especially the mass production of valuable human proteins and xenotransplantation. In the 21st century animal biotechnology will further contribute to welfare of human being.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.95-98
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• 2013

Following construction of expression vectors in Escherichia coli, a new procedure involving two-step column purifications with a Fast Performance Liquid Chromatography System was developed for purification of the oxygenase component of alkylphenol hydroxylase of Pseudomonas alkylphenolia. From 50 g wet cake of recombinant E. coli BL21(DE3)(pJJPMO2) cells, 110 mg of pure protein in a heterodimeric form containing a stoichiometric amount of iron were obtained and it exhibited a specific activity of 147 nmole/min/mg.

• Plant Resources
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• v.4 no.3
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• pp.181-188
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• 2001

We tried to introduce two forsythia genes related in lignan biosynthesis, dirigent protein and pinoresinol/lariciresinol (Ph) reductase, into potatoes for accumulation of lignans in transgenic potatoes. We made binary vectors overexpressing dirigent protein gene and P/L reductase gene driven by a CaMV35S promoter and transformed into potatoes via Agrobacterium mediated transformation. And in order to control the metabolic flux of lignan biosynthesis pathway, we tried to inhibit chalcone synthase genes of potatoes by antisense inhibition technique also. We tried to use PCR screening method for selection of transgenic plants of different vectors. We tried to determine and compare lignan contents from different transgenic potato lines.

• International Journal of Control, Automation, and Systems
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• v.5 no.2
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• pp.111-116
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• 2007

The design problem of partial pole assignment (PPA) in two classes of frequency-domain MIMO models by constant gain feedback is investigated in this paper. Its aim is to design a constant gain feedback which changes only a subset of the open-loop eigenvalues, while the rest of them are kept unchanged in the closed-loop system. A near general parametric expression for the feedback gain matrix in term of a set of design parameter vectors and the set of the closed-loop poles, and a simple parametric approach for solving the proposed problem are presented. The set of poles do not need to be previously prescribed, and can be set undetermined and treated together with the set of parametric vectors as degrees of design freedom provided by the approach. An illustrative example shows that the proposed parametric method is simple and effective.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.543-547
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• 2002

DNA vaccines use eukaryote expression vectors to produce immunizing proteins in the vaccinated host and it represents a novel approach to vaccine and immuno-therapeutic development. We constructed a 2.9 kb compact plasmid vector (pVAC) which contains CMV promoter, polycloning site, BGH poly A terminator, ampicillin resistance gene and PBR322 origin. Enriched unmathlyated CpG motifs have introduced into pVAC-ISS1 and pVAC-ISS2 which are derived from pVAC for enhancing Thl responses. These plasmid DNAs rapidly induced interleukin 6 secretion in vivo. It is expected that these vectors will contribute to the DNA inoculation against infectious disease and various cancers without adjuvant.

• YAKHAK HOEJI
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• v.47 no.3
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• pp.176-179
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• 2003

A library of unlimited number of novel lectins with diverse specificities has been previously generated by randomly mutating the carbohydrate-recognition domain of Maackia amurensis hemagglutinin (MAH). To establish the experimental environment capable of selecting high affinity mutant lectins in E. coli, phage display system was adapted. Carbohydrate binding capacity of two phagemid vectors, pComb3 and pComb8 displaying wild-type MAH lectin was assessed. Specific bindings of pComb3 and pComb8 phages expressing w.t. MAH to affinity-purified polyclonal anti-MAH antibody and to glycophorin was demonstrated. Both phages also showed strong hemagglutinating activity to intact but not sialidase-treated human erythrocytes, which is consistent to the specificity of native MAH. Taken together, two different phage display vectors successfully allowed the expression of active MAH as a fusion protein on the surface of bacteriophage, which will lead to preparation of unique plant lectins with high affinity toward a variety of carbohydrate chains.