• Journal of Sericultural and Entomological Science
• /
• v.39 no.1
• /
• pp.36-43
• /
• 1997

In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

• Journal of the Korea Institute of Information and Communication Engineering
• /
• v.12 no.9
• /
• pp.1547-1553
• /
• 2008

This paper presents a method that generates various facial expressions of vector graphic character by using the simplified principle component vector. First, we analyze principle components to the nine facial expression(astonished, delighted, etc.) redefined based on Russell's internal emotion state. From this, we find principle component vector having the biggest effect on the character's facial feature and expression and generate the facial expression by using that. Also we create natural intermediate characters and expressions by interpolating weighting values to character's feature and expression. We can save memory space considerably, and create intermediate expressions with a small computation. Hence the performance of character generation system can be considerably improved in web, mobile service and game that real time control is required.

• Journal of Microbiology and Biotechnology
• /
• v.23 no.3
• /
• pp.322-328
• /
• 2013

In this study, we show the expression and purification of the human recombinant farnesoid X receptor (FXR)- ligand binding domain (LBD) protein in E. coli using a double cistronic vector, pACYCDuet-1, as a soluble form. We describe here the expression and characterization of a biologically active $FXR-LBD_{(248-476)}$. When expressed in the influence of bacterial promoters ($P_{T7}$ and $P_{Tac}$) of the single cistronic expression vectors, the human recombinant $FXR-LBD_{(248-476)}$ was found to be totally insoluble. However, by using a double cistronic expression vector, we were able to obtain the human recombinant $FXR-LBD_{(248-476)}$ in a soluble form. To allow for biological activities, we have subcloned into the pACYCDuet-1 vector, expressed in E. coli cells at some optimized conditions, and purified and characterized the human recombinant active $FXR-LBD_{(248-476)}$ proteins using the fluorescence polarization assay. This suggests that the expression of FXR-LBD in a double cistronic vector improves its solubility and probably assists its correct folding for the biologically active form of the proteins. We suggest that this may represent a new approach to high expression of other nuclear receptors and may be useful as well for other classes of heterodimeric protein partners.

• Journal of Microbiology and Biotechnology
• /
• v.1 no.1
• /
• pp.37-44
• /
• 1991

A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the vector, a useful DNA fragment, 319 bp EcoRI fragment, having strong promoter activity has been cloned from Bacillus subtills chromosomal DNA. Selection was based on chloramphenicol resistance which is dependent upon the introduction of DNA fragments allowing expression of a chloramphenicol acetyl transferase gene. The nucleotide sequence of the 319 bp fragment has been determined and the putative -35 and -10 region, ribosome binding site, and ATG initiation codon were observed. This promoter was named EB promoter and the resultant plasmid which can be used as an expression vector was named pEBP313. The crystal protein gene from B. thuringiensis subsp. kurstaki HD-73 was cloned downstream from the EB promoter without its own promoter. When the resultant plasmid, pBT313, was introduced into Escherichia coli and B. subtilis, efficient synthesis of crystal protein was observed in both cells, and the cp gene expression in B. subtilis begins early in the vegetative phase. The cell extracts from both clones were toxic to Hyphantria cunea larvae.

• Journal of Microbiology and Biotechnology
• /
• v.8 no.6
• /
• pp.625-630
• /
• 1998

Using a modified yeast secretory expression vector, $\alpha$-amylase of Schwanniomyces occidentalis was produced from Saccharomyces cerevisiae. The expression vector contains the a-amylase gene (AMY) harboring its own promoter without the regulatory region and the adenine base at the -3 position from the ATG start codon, its own signal sequence, CYC1 transcription terminator, and SV40 enhancer. The expressed $\alpha$-amylase activity from cells carrying the plasmid was approximately 26 times higher than that from the cells harboring an unmodified plasmid. When Saccharomyces diastaticus was transformed with this modified vector, a 2.5 times higher level of amylolytic activity than that from Sch. occidentalis was observed.

• Journal of information and communication convergence engineering
• /
• v.17 no.1
• /
• pp.14-20
• /
• 2019

Currently, microarray gene expression data take advantage of the sufficient classification of cancers, which addresses the problems relating to cancer causes and treatment regimens. However, the sample size of gene expression data is often restricted, because the price of microarray technology on studies in humans is high. We propose enhancing the gene expression classification of support vector machines with generative adversarial networks (GAN-SVMs). A GAN that generates new data from original training datasets was implemented. The GAN was used in conjunction with nonlinear SVMs that efficiently classify gene expression data. Numerical test results on 20 low-sample-size and very high-dimensional microarray gene expression datasets from the Kent Ridge Biomedical and Array Expression repositories indicate that the model is more accurate than state-of-the-art classifying models.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
• /
• 2014.10a
• /
• pp.977-980
• /
• 2014

A baculovirus vector systems including genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were transfected into human foreskin fibroblast cells and various tissues and investigated gene transfer and expression of these vector systems with control vectors. From the study, these recombinant baculovirus vector systems were more effective and safe than control vector in view of gene transfer and expression.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.1
• /
• pp.507-510
• /
• 2013

Background: More and more research indicate that the immediately early response gene 3 (IER3) is involved inmany biological provesses, such as apoptosis and immunoreaction, as well as viral infection, tumorigenesis and tumour progression. Methods: Here we describe the construction of an eukaryotic expression vector containing IER3 gene and its expression in A549 cells as assessed through fluorescence microscopyand Western-blotting. Results: Fluorescence detection displayed that GFP in cytoplasm was high during 48 and 72 hours post-transfection. In addition, Western blotting showed significant increase in IER3 gene expression in the transfected cells compared with controls. Conclusion: The recombinate plasmid expression vector was constructed successfully, which may provide a basis for further exploration of function of IER3 in lung cancer.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.9
• /
• pp.1362-1368
• /
• 2006

A new constitutive episomal expression vector, pGAPZ-E, was constructed and used for initial screening of eukaryotic target gene expression in Pichia pastoris. Two reporter genes such as beta-galactosidase gene and GFPuv gene were overexpressed in P. pastoris. The expression level of the episomal pGAPZ-E strain was higher than that of the integrated form when the beta-galactosidase gene was used as the reporter gene in P. pastoris X33. The avoiding of both the integration procedure and an induction step simplified the overall screening process for eukaryotic target gene expression in P. pastoris. Nine human protein targets from the Core 50, family of Northeast Structural Genomics Consortium (http://www.nesg.org), which were intractable when expressed in E. coli, were subjected to rapid screening for soluble expression in P. pastoris. HR547, HR919, and HR1697 human proteins, which had previously been found to express poorly or to be insoluble in E. coli, expressed in soluble form in P. pastoris. Therefore, the new episomal GAP promoter vector provides a convenient and alternative system for high-throughput screening of eukaryotic protein expression in P. pastoris.

• Microbiology and Biotechnology Letters
• /
• v.20 no.6
• /
• pp.637-641
• /
• 1992

The promoter regions from chromosomal DNA of Bacillus sp. SSA3 which is responsible for fermentation of Korean traditional soy sauce, were cloned for construction of expression vector of Bacills sp. SSA3. Recombinant plasmids were constructed by insertion of HindIIl-cleaved Bacillus sp. SSA3 chromosomal DNA fragments in front of the CAT gene of pGR71 plasmid and B-galactosidase gene of pUC18 plasmid. 6 recombinant plasmids were isolated from chloramphenicol resistant E. coli JM109 clones. All these plasmids were found to have promoter activity in Bacills sp. SSA3 and E. coli JM109. When these 6 clones of Bacills sp. SSA3 were cultivated in LB agar medium supplemented with 10% NaCI. fused CAT gene expression of 4 clones was significantly decreased in common. But the others were poorly inhibited.