• Title/Summary/Keyword: vector manipulation

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Ear Recognition by Major Axis and Complex Vector Manipulation

  • Su, Ching-Liang
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.11 no.3
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    • pp.1650-1669
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    • 2017
  • In this study, each pixel in an ear is used as a centroid to generate a cake. Subsequently the major axis length of this cake is computed and obtained. This obtained major axis length serves as a feature to recognize an ear. Later, the ear hole is used as a centroid and a 16-circle template is generated to extract the major axis lengths of the ear. The 16-circle template extracted signals are used to recognize an ear. In the next step, a ring-to-line mapping technique is used to map these major axis lengths to several straight-line signals. Next, the complex plane vector computing technique is used to determine the similarity of these major axis lengths, whereby a solution to the image-rotating problem is achieved. The aforementioned extracted signals are also compared to the ones that are extracted from its neighboring pixels, whereby solving the image-shifting problem. The algorithm developed in this study can precisely identify an ear image by solving the image rotation and image shifting problems.

A New Bicistronic Fragmentation Vector for Manipulation and Analysis of Functional Yeast Artificial Chromosomes (YACs) (Yeast Artificial Chromosome의 효율적인 조작과 분석을 위한 새로운 Bicistronic Fragmentation Vector의 개발에 관한 연구)

  • 임향숙;최주연;김인경;강성만;성영모
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.28-34
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    • 1999
  • Fragmentation vectors are used to analyze function and genomic structure of a gene of interest by creating deletion derivatives of large fragments of genomic DNA cloned as yeast artificial chromosomes (YACs). Herein, we developed a new hicistronic fragmentation vector that contains internal ribosomal entry sile (IRES) of encephalomyocarditis vin~s (EMCV) and $\beta$-galactosidase as a reporter gene. This vector system provides a novcl loo1 to analyze expression patterns of a gene of interest due to simultaneous expression of a target gene as well as $\beta$-galactosidase driven from a single message. In addition, the bicistronic fragmentation vector contains four rare-cutting restriction enzyme sites in the polycloning sites which can be used to conveniently insert any kinds of genes and therefore facilitates targeting DNA scgments into YAC by means of homologous recombination. This approach establishes a paradigm for manipulation of mammalian DNA segments and characterization of expression and regulatory regions of mammalian gene cloned as YAC.

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Construction of rhizobium-E. coli shuttle vector using replication and mobilization function of indigenous multicopy plasmid from rhizobium (Rhizobium muliticopy plasmid의 복제 및 이주 기능을 이용한 rhizobium-E. coli shuttle vector 구축)

  • 조무제;신평균;최영주;강규영;윤한대
    • Korean Journal of Microbiology
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    • v.27 no.2
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    • pp.92-97
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    • 1989
  • the vector, pGUR19, for Rhizobium gene manipulation, was constructed by combining the replication and mobilization function of indigenous multicopy plasmid from Acacia(Robinia pseudoacacia L.) Rhizobia sp86 with E. coli cloning vehicle, pBR322. The vector could be efficiently mobilized by RP4 tra function incorporated into chromosome of E. coli named SM10 and efficiently transferred to various gram negative hosts including Rhizobium and Afrobacterium by transformation. Mobilization frequency of the constructed vector was ranged from $1.2\times 10^{-2}$ (E.coli HB 101) to $4.6\times 10^{-4}$ (A. tumefaciens 15955) and transformation frequency was ranged from $5.4\times 10^{-7}$(E. coli HB101) to $1.2\times 10^{-10}$ (A. tumefaciens 15955). The vector, pGUR19, was stably replicated and maintained in a variety of Rhizobium and Agrobacterium.

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Identification of Gene Locus by the Somatic Cell Hybridization in Chicken (체세포 융합에 의한 닭의 유전인자구명에 관한 연구)

  • 정익정
    • Korean Journal of Poultry Science
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    • v.16 no.1
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    • pp.1-8
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    • 1989
  • This experiment was conducted to improve the performance of chickens by the precise separation and analysis of chromosomes which are integrated genetic materials, and by the use of gene manipulation techniques. Following are the main results obtained. 1. When the chromosomes were separated through the leucocyte culture and analyzed by Giemsa banding techniques (especially by the method in which 20 layers of banding patterns could be found in chromosome #1), the normal Patterns of chromosomes #l-9 and sex chromosomes, and the location of constitutive heterochromatin without any gene activities in all chromosomes were discovered. 2. To utilize the primodial germ cells (PGC) as the genetic vector which is one of the most important gene manipulation techniques, PGC's from triploid were transplanted to normal host embryos. Since the donor PGC's(3n) were found in the gonads of growing host embryos gene manipulation in poultry using PGC's, seemed to be possible.

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A New Reporter Vector System Based on Flow-Cytometry to Detect Promoter Activity

  • Jung, Sun-Do;Choi, Ji-Hye;Hong, Chang-Wan;Lee, Hyun-Ji;Park, Yoon-Kyung;Shin, Jung-Hoon;Park, Jae-Won;Park, Se-Ho
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.243-247
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    • 2009
  • In this study, we report the development of a new dual reporter vector system for the analysis of promoter activity. This system employs green fluorescence emitting protein, EGFP, as a reporter, and uses red fluorescence emitting protein, DsRed, as a transfection control in a single vector. The expression of those two proteins can be readily detected via flow cytometry in a single analysis, with no need for any further manipulation after transfection. As this system allows for the simultaneous detection of both the control and reporter proteins in the same cells, only transfected cells which express the control protein, DsRed, can be subjected to promoter activity analysis, via the gating out of all un-transfected cells. This results in a dramatic increase in the promoter activity detection sensitivity. This novel reporter vector system should prove to be a simple and efficient method for the analysis of promoter activity.

Development of Host-Vector Systems for Lactic Acid Bacteria (유산균의 Host-Vector System 개발)

  • 윤성식;김창민
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.1-11
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    • 2001
  • Lactic acid bacteria (LAB) are widely used for various food fermentation. With the recent advances in modern biotechnology, a variety of bio-products with the high economic values have been produced using microorganisms. For molecular cloning and expression studies on the gene of interest, E. coli has been widely used mainly because vector systems are fully developed. Most plasmid vectors currently used for E, coli carry antibiotic-resistant markers. As it is generally believed that the antibiotic resistance markers are potentially transferred to other bacteria, application of the plasmid vectors carrying antibiotic resistance genes as selection markers should be avoided, especially for human consump-tion. By contrast, as LAB have some desirable traits such that the they are GRAS(generally recognized as safe), able to secrete gene products out of cell, and their low protease activities, they are regarded as an ideal organism for the genetic manipulation, including cloning and expression of homologous and heterologous genes. However, the vec-tor systems established for LAB are stil insufficient to over-produce gene products, stably, limiting the use of these organisms for industrial applications. For a past decade, the two popular plasmid vectors, pAM$\beta$1 of Streptococcus faecalis and pGK12 theB. subtilis-E. coli shuttle vector derived from pWV01 of Lactococcus lactis ssp. cremoris wg 2, were most widely used to construct efficient chimeric vectors to be stably maintained in many industrial strains of LAB. Currently, non-antibiotic markers such as nisin resistance($Nis^{r}$ ) are explored for selecting recombi-nant clone. In addition, a gene encoding S-layer protein, slp/A, on bacterial cell wall was successfully recombined with the proper LAB vectors LAB vectors for excretion of the heterologous gene product from LAB Many food-grade host vec-tor systems were successfully developed, which allowed stable integration of multiple plasmid copies in the vec-mosome of LAB. More recently, an integration vector system based on the site-specific integration apparatus of temperate lactococcal bacteriophage, containing the integrase gene(int) and phage attachment site(attP), was pub-lished. In conclusion, when various vector system, which are maintain stably and expressed strongly in LAB, are developed, lost of such food products as enzymes, pharmaceuticals, bioactive food ingredients for human consump-tion would be produced at a full scale in LAB.

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An Optimization Strategy for Vector Spatial Data Transmission onover the Internet (인터넷을 통한 벡터 공간 데이타의 효율적 전송을 위한 최적화 기법)

  • Liang Chen;Chung-Ho Lee;Hae-Young Bae
    • Journal of KIISE:Databases
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    • v.30 no.3
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    • pp.273-285
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    • 2003
  • Generally, vector spatial data, with richer information than raster spatial data enabledata, enables a mere flexible and effective manipulation of the data sets. However, one of challenges against the publication of vector spatial information on the Internet is the efficient transmission of the big and complex vector spatial datadata, which is both large and complex, across the narrow-bandwidth of the Internet. This paper proposes a new transmission method, namely, the Scale-Dependent Transmission method, with the purpose of improving the efficiency of vector spatial data transmission on the narrow-bandwidthacross the Internet. Simply put, its nam idea is “Transmit what can be seen””. Scale is regarded as a factor naturally associated with spatial features so that not all features are visible to users at a certain scale. With the aid of the Wavelet-Wavelet-based Map Generalization Algorithm, the proposed method filters out invisible features from spatial objects according to the display scale and then to transmit onlytransmits only the visible features as athe final answer for an individual operation. Experiments show that the response times ofan individual operation has been reducedoperations were substantially by the usage of reduced when using the proposed method.

Development of New Vector Systems as Genetic Tools Applicable to Mycobacteria (Mycobacteria에 적용 가능한 genetic tool로서의 새로운 vector system 개발)

  • Jeong, Ji-A;Lee, Ha-Na;Ko, In-Jeong;Oh, Jeong-Il
    • Journal of Life Science
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    • v.23 no.2
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    • pp.290-298
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    • 2013
  • The genus Mycobacterium includes crucial animal and human pathogens such as Mycobacterium tuberculosis, Mycobacterium leprae, and Mycobacterium bovis. Although it is important to understand the genetic basis for their virulence and persistence in host, genetic analysis in mycobacteria was hampered by a lack of sufficient genetic tools. Therefore, many functional vectors as molecular genetic tools have been designed for understanding mycobacterial biology, and the application of these tools to mycobacteria has accelerated the study of mechanisms involved in virulence and gene expression. To overcome the pre-existing problems in genetic manipulation of mycobacteria, this paper reports new vector systems as effective genetic tools in Mycobacterium smegmatis. Three vectors were developed; pKOTs is a suicide vector for mutagenesis containing a temperature-sensitive replication origin (TSRO) and the sacB gene encoding levansucrase as a counterselectable marker. pMV306lacZ is an integrative lacZ transcriptional fusion vector that can be inserted into chromosomal DNA by site-specific recombination. pTnMod-OKmTs is a minitransposon vector harboring the TSRO that can be used in random mutagenesis. It was demonstrated in this study that these vectors effectively worked in M. smegmatis. The vector systems reported here are expected to successfully applicable to future research of mycobacterial molecular genetics.

Manipulation of Hepatitis B Viral DNA for Generating Transgenic Mice

  • Kim, Seung-Hee;Park, Sang-Ho;Kim, Tae-Gyun;Lee, Song-Deuk;Aree Moon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.178-178
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    • 1996
  • Hepatitis B virus (HBV) infection is one of the serious problems in Southeast Asia including Korea because it causes chronic hepatitis, which can easily be transformed In fatal conditions such as cirrhosis and hepatoma. Even though lots of informations on structural characteristics and gene expression mechanisms have been accumulated, the mechanism for HBV-induced hepatocellular injury which is believed to be the consequences of the immunological response is not well understood. In order tn perform immunopathological studies for prevention and treatment of HBV infection, we designed transgenic mice as a disease model which can mimic HBV infection, In this study, a promoter-HBV DNA fragment for the preparation of HBV transgenic mice has been constructed. To add a proper enzyme site on 5' end of HBV gene, total HBV (subtype adr) gene was inserted into BamHI site of pBluescript SK vector and reextracted by PstI-SacI treatment A liver-specific promoter, rat ${\alpha}$ 2u globulin gene promoter, was insrted to pBluescript SK vector and reextracted by BamHI-PstI treatment, Promoter-HBV DNA was constructed by ligation of two fragments using identical PstI sites. For large scale production of promoter-HBV DNA, it was inserted to BamHI-SacI site of pBluescript SK vector.

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Novel Vectors for the Convenient Cloning and Expression of In Vivo Biotinylated Proteins in Escherichia coli

  • Cho, Eun-Wie;Park, Jung-Hyun;Na, Shin-Young;Kim, Kil-Lyong
    • BMB Reports
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    • v.32 no.5
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    • pp.497-501
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    • 1999
  • Biotinylation of recombinant proteins is a powerful tool for the detection and analysis of proteins of interest in a large variety of assay systems. The recent development of in vivo biotinylation techniques in E. coli has opened new possibilities for the production of site-specifically biotinylated proteins without the need for further manipulation after the isolation of the recombinantly expressed proteins. In the present study, a novel vector set was generated which allows the convenient cloning and expression of proteins of interest fused with an N-terminal in vivo biotinylated thioredoxin (TRX) protein. These vectors were derived from the previously reported pBIOTRX vector into which was incorporated part of the pBluescript II+phagemid multiple cloning site (MCS), amplified by PCR using a pair of sophisticated oligonucleotide primers. The functionality of these novel vectors was examined in this system by recombinant expression of rat transforming growth factor-$\beta$. Western-blot analysis using TRX-specific antibodies or peroxidase-conjugated streptavidin confirmed the successful induction of the fusion protein and the in vivo conjugation of biotin molecules, respectively. The convenience of molecular subcloning provided by the MCS and the effective in vivo biotinylation of proteins of interest makes this novel vector set an interesting alternative for the production of biotinylated proteins.

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