• BMB Reports
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• v.39 no.3
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• pp.329-334
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• 2006

RNAi (RNA interference) has become a popular means of knocking down a specific gene in vivo. The most common approach involves the use of chemically synthesized short interfering RNAs (siRNAs), which are relatively easy and fast to use, but which are costly and have only transient effects. These limitations can be overcome by using short hairpin RNA (shRNA) expression vectors. However, current methods of generating shRNA expression vectors require either the synthesis of long (50-70 nt) costly oligonucleotides or multi-step processes. To overcome this drawback, we have developed a one-step short-oligonucleotides-based method with preparation costs of only 15% of those of the conventional methods used to obtain essentially the same DNA fragment encoding shRNA. Sequences containing 19 bases homologous to target genes were synthesized as 17- and 31-nt DNA oligonucleotides and used to construct shRNA expression vectors. Using these plasmids, we were able to effectively silence target genes. Because our method relies on the onestep ligation of short oligonucleotides, it is simple, less error-prone, and economical.

• The Mathematical Education
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• v.56 no.1
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• pp.63-80
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• 2017

By analysing the examination papers from third grade high school students, we classified the errors occurred in the problem solving process of high school 'Geometry and Vectors' into several types. There are five main types - (A)Insufficient Content Knowledge, (B)Wrong Method, (C)Logical Invalidity, (D)Unskilled Expression and (E)Interference.. Type A and B lead to an incorrect answer, and type C and D cannot be distinguished by multiple-choice or closed answer questions. Some of these types are classified into subtypes - (B1)Incompletion, (B2)Omitted Condition, (B3)Incorrect Calculation, (C1)Non-reasoning, (C2)Insufficient Reasoning, (C3)Illogical Process, (D1)Arbitrary Symbol, (D2)Using a Character Without Explanation, (D3) Visual Dependence, (D4)Symbol Incorrectly Used, (D5)Ambiguous Expression. Based on the these types of errors, answers of each problem was analysed in detail, and proper ways to correct or prevent these errors were suggested case by case. When problems that were used in the periodical test were given again in descriptive forms, 67% of the students tried to answer, and 14% described flawlessly, despite that the percentage of correct answers were higher than 40% when given in multiple-choice form. 34% of the students who tried to answer have failed to have logical validity. 37% of the students who tried to answer didn't have enough skill to express. In lessons on curves of secondary degree, teachers should be aware of several issues. Students are easily confused between 'focus' and 'vertex', and between 'components of a vector' and 'coordinates of a point'. Students often use an undefined expression when mentioning a parallel translation. When using a character, students have to make sure to define it precisely, to prevent the students from making errors and to make them express in correct ways.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.550-556
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• 1995

The c-myc proto-oncogene is Involved In the control of normal cell proliferation and differentiation of many cell lineages. Although it has heen suggested that c-myc may play an important role in the mammalian early development, it Is unclear whether the embryonic c-myc mRNA is originated from zygotic gene expression or stored maternal message. Thus, we have construded expression vectors, In which the 5, flanking sequences including c-myc promoter region and a large non-coding exon I are fused 'sith E. coli lacZ gene that encedes $\beta$-galactosldase as a reporter. As c-myc exon I contains a modulatory sequence, we designed t, vo types of vectors (pcmyc.Gall and pcmyc-Ga12) to examine the role of exon I in c-myc expression. The former contains the complete exon I and the later has a deletion in 40 bp of modulator sequence located In the exon I of c-myc These vectors were microInjected into fertilized one-cell embryos and $\beta$-galactosidase activity was examined by X-gal staining during early embryogenesis. $\beta$-galactosidase activity derived from c-myc promoter was decreased at two-cell stage. The expression level directed by pcmyc- Ga12 was similar to that of pcmyc-Gal1, indicating that the medulatory sequence in exon I may not be Involved at least In the regulation of embryonic c-myc expression. In summary, the present study indicates that the c-myc promoter is functional at the early stage embryo, and the regulation of c-myc expression is under the control of "zygotic" clock of preimplantation mouse embryos.e embryos.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.70-73
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• 2001

Isopentenyl diphosphate (IPP) is the common, five-carbon building block in the biosynthesis of all isoprenoids. IPP in Escherichia coli is synthesized through the non-mevalonate pathway. The first reaction of IPP biosynthesis in E. coli is the formation of 1-deoxy-D-xylulose-5-phosphate(DXP), catalyzed by DXP synthase and encoded by dxs. The second reaction in the pathway is the reduction of DXP to 2-C-methyl-D-erythritol-4-phosphate, catalyzed by DXP reductoismerase and encoded by dxr. To determine if one of more of the reactions in the non-mevalonate pathway controlled flux to IPP, dxs and dxr were placed on several expression vectors under the control of three different promoters and transformed into three E. coli strains ($DH5{\alpha}$, XL1-Blue, and JM101) that had been engineered to produce lycopene, a kind of isoprenoids. Lycopene production was improved significantly in strains transformed with the dex expression vectors. At arabinose concentrations between 0 and 1.33 mM, cells expressiong both dxs and from $P_{BAD}$ on a midium-copy plasmid produced 1.4 -2.0 times more lycopene than cells expressing dxs only. However, at higher arabinose concentrations lycopene production in cell expressing both dxs and dxr was lower than in cells expression dxs only. A comparison of the three E. coli strains trasfomed with the arabinose-inducible dxs on a medium-copy plasmid revealed that lycopene production was highest in XL1-Blue.

• BMB Reports
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• v.28 no.3
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• pp.216-220
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• 1995

cDNA for human cholesteryl ester transfer protein (CETP), a potent atherogenic plasma protein that redistributes the neutral lipids among lipoproteins, was expressed in recombinant vaccinia virus-infected cells (CV-1). Two insertion vectors regulated by different promoters were constructed. The vectors were introduced into human thymidine kinase-negative ($TK^-$) 1438 cells infected with wild-type vaccinia virus (WR strain). Recombinant viruses were selected with 5-bromodeoxyuridine (BUdR) and X-gal and identified with DNA dot blot analysis (vSC11-CETP and vTM1-CETP). The CETP cDNA insert in the recombinant vaccinia virus genome was identified by Southern blot analysis. Transcription of CETP cDNA in CV-1 cells infected with recombinant vaccinia virus was monitored by Northern blot analysis using the CETP cDNA as a probe. Positive signals were detected at 1.8 kb in cells infected with vSC11-CETP and at 2.3 kb in cells infected with vTM1-CETP. The recombinant vaccinia virus-infected CV-1 cells were shown to produce functional CETP when the culture medium was subjected to the CETP assay.

• Journal of Korea Multimedia Society
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• v.12 no.8
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• pp.1155-1163
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• 2009

This paper proposes an automatic facial expression generation system of vector graphic character using gaussian process model. Proposed method extracts the main feature vectors from twenty-six facial data of character redefined based on Russell's internal emotion state. Also by using new gaussian process model, SGPLVM, we find low-dimensional feature data from extracted high-dimensional feature vectors, and learn probability distribution function (PDF). All parameters of PDF are estimated by maximization the likelihood of learned expression data, and these are used to select wanted facial expressions on two-dimensional space in real time. As a result of simulation, we confirm that proposed facial expression generation tool is working in the small facial expression datasets and can generate various facial expressions without prior knowledge about relation between facial expression and emotion.

• Journal of the Korean Institute of Intelligent Systems
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• v.16 no.6
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• pp.759-765
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• 2006

In this paper, it is proposed an effective face expression recognition LDA mixed mode using a triangularity membership fuzzy function and wavelet basis. The proposal algorithm gets performs the optimal image, fuzzy wavelet algorithm and Expression recognition is consisted of face characteristic detection step and face Expression recognition step. This paper could applied to the PCA and LDA in using some simple strategies and also compares and analyzes the performance of the LDA mixed model which is combined and the facial expression recognition based on PCA and LDA. The LDA mixed model is represented by the PCA and the LDA approaches. And then we calculate the distance of vectors dPCA, dLDA from all fates in the database. Last, the two vectors are combined according to a given combination rule and the final decision is made by NNPC. In a result, we could showed the superior the LDA mixed model can be than the conventional algorithm.

• Mycobiology
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• v.45 no.2
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• pp.84-89
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• 2017

Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii.

• The Transactions of the Korean Institute of Electrical Engineers D
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• v.50 no.3
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• pp.149-157
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• 2001

In the video retrieval system, visual clues of still images and motion information of video are employed as feature vectors. We generate the temporal textures to express the motion information whose properties are simple expression, easy to compute. We make those temporal textures of wavelet coefficients to express motion information, M components. Then, temporal texture feature vectors are extracted using spatial texture feature vectors, i.e. spatial gray-level dependence. Also, motion amount and motion centroid are computed from temporal textures. Motion trajectories provide the most important information for expressing the motion property. In our modeling system, we can extract the main motion trajectory from the temporal textures.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2019.05a
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• pp.533-536
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• 2019

Recombinant baculoviruses are widely used to express heterologous genes in cultured insect cells. Recombinant baculoviruses can serve as gene-transfer vectors for expression of recombinant proteins in a wide range of mammalian cell types. Baculovirus system has significant benefits in view of safety, large-scale, and high level of gene expression. In this study, baculoviral vectors which were reconstructed from pOPINEneo-3C-GFP vector, were recombined with cytomegalovirus (CMV) promoter, green fluorescent protein (GFP), and p53 with NcoI and XhoI. These recombinant vectors were infected with various cells and cell lines. The baculovirus vector thus developed was analyzed by comparing the metastasis and expression of the recombinant genes with conventional vectors. These results suggest that the baculovirus vector has higher efficiency in metastasis and expression than the control vector. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).