• Title/Summary/Keyword: integration vector

Search Result 298, Processing Time 0.032 seconds

Development of a Multicopy Integration Vector in Yarrowia lipolytica (Yarrowia lipolytica의 Multicopy Integration Vector 개발)

  • Kim, Jeong-Yoon;Woo, Moon-Hee;Ryu, Dewey D.Y.
    • Microbiology and Biotechnology Letters
    • /
    • v.23 no.5
    • /
    • pp.536-543
    • /
    • 1995
  • Multicopy integration vector is a very useful vector system in that they can be integrated into chromosomal DNA in several copies and stably maintained under non-selective conditions. To develop a multicopy integration vector system in the yeast Yarrowia lipolytica, P-type ribosomal DNA was cloned from Y lipolytica. A HindIII-BglII fragment of the cloned rDNA and a promoterless URA3 gene were inserted into pGEM1, generating multicopy integration vectors, pMIYL-1 and pMIYL-2. The rDNA fragment is for targeted homologous recombination between the vector and the chromosomal DNA of Y. lipolytica, and the promoterless URA3 gene is a defective selection marker for inducing multicopy integration. pMIYL-1 and pMIYL-2 have an unique restriction enzyme site, KpnI, and two unique restriction enzyme sites, KpnI and EcoRI, repectively, which can be used for targeting of the vectors into the rDNA of Y. lipolytica chromosomal DNA. After transformation of the vectors into Y. lipolytica, copy number and stability were analyzed by Southern hybridization. The vectors were found to be present in less than 5 copies per cell and were stably maintained during growth in non-selective media.

  • PDF

Development of a Recombinant Streptomyces griseus with sprA and sprB Genes for Proteolytic Enzyme Production (Streptomyces griseus IFO13350 유래 sprA 및 sprB 유전자를 이용한 Pretense 생산균주 개발)

  • Hwang Ji-Hwan;Lee Chang-Kwon;Lee Kang-Mu;Jo Byoung-Kee;Park Hae-Ryong;Hwang Yong-Il
    • Korean Journal of Microbiology
    • /
    • v.41 no.1
    • /
    • pp.87-92
    • /
    • 2005
  • Pronase, a protease produced for commercial purpose by Streptomyces griseus, was composed of serine protease, alkaline protease, aminopeptidase and carboxypeptidase complex, and it has been widely used as anti-inflammatory drugs for human therapy. In this study, we developed a new integration vector, pHJ101 derived from pSET152, containing strong promoter, ermE, to overexpress a certain protease gene. Specific PCR primers for cloning of sprA (a gene for S. griseus protease A) and sprB (a gene for S. griseus protease B) genes were designed from the basis of nucleotide sequence in databases and amplified by PCR. Plasmid pHJ201 and pHJ202 were constructed by inserting of amplified each gene in a vector pHJ101. S. griseus HA and S. griseus HB were respectively obtained by conjugal process of a parent strain, S. griseus IFO 13350 with the recombinant Escherichia coli harboring plasmid pHJ201 or pHJ202. When protease activity was measured in flask cultivation, produced protease levels of S. griseus HA and S. griseus HB increased about 5.3 times and 5 times, respectively, more than that of parent strain. And, the constructed integrating plasmid pHJ101 was applicable for overexpression of a certain gene in Streptomyces sp.

Performance Evaluation of Vector Tracking Loop Based Receiver for GPS Anti-Jamming Environment (GPS 교란 환경에서 벡터추적루프 기반 수신기 성능평가)

  • Song, Jong-Hwa;Im, Sung-Hyuck;Jee, Gyu-In
    • Journal of Institute of Control, Robotics and Systems
    • /
    • v.19 no.2
    • /
    • pp.152-157
    • /
    • 2013
  • In this paper, we represent the implementation and performance analysis of vector tracking loop based GPS receiver for jamming environment. The vector tracking loop navigation performance is compared by simulation with conventional tracking loop. The simulation results shows that vector tracking loop is more robust than conventional tracking loop in jamming environment. The vector tracking loop can gain 2dB in jamming performance capability over a conventional GPS receiver. Also, Anti-jamming performance of INS Doppler aiding and deep integration method are compared.

Development of a Food-Grade Integration Vector for Heterologous Gene Expression and Protein Secretion in Lactococcus lactis

  • Jeong, Do-Won;Lee, Jong-Hoon;Kim, Kyoung-Heon;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.11
    • /
    • pp.1799-1808
    • /
    • 2006
  • A food-grade integration vector based on site-specific recombination was constructed. The 5.7-kb vector, pIMA20, contained an integrase gene and a phage attachment site originating from bacteriophage A2, with the ${\alpha}$-galactosidase gene from Lactobacillus plantarum KCTC 3104 as a selection marker. pIMA20 was also equipped with a controllable promoter of nisA ($P_{nisA}$) and a signal peptide-encoding sequence of usp45 ($SP_{usp45}$) for the production and secretion of foreign proteins. pIMA20 and its derivatives mediated site-specific integration into the attB-like site on the Lactococcus lactis NZ9800 chromosome. The vector-integrated recombinant lactococci were easily detected by the appearance of blue colonies on a medium containing $X-{\alpha}-gal$ and also by their ability to grow on a medium containing melibiose as the sole carbon source. Recombinant lactococci maintained these traits in the absence of selection pressure during 100 generations. The ${\alpha}-amylase$ gene from Bacillus licheniformis, lacking a signal peptide-encoding. sequence, was inserted downstream of $P_{nisA}\;and\;SP_{usp45}$ in pIMA20, and the plasmid was integrated into the L. lactis chromosome. ${\alpha}-Amylase$ was successfully produced and secreted by the recombinant L. lactis, controlled by the addition and concentration of nisin.

SCALED VISUAL CURVATURE AND VISUAL FRENET FRAME FOR SPACE CURVES

  • Jeon, Myungjin
    • Journal of the Chungcheong Mathematical Society
    • /
    • v.34 no.1
    • /
    • pp.37-53
    • /
    • 2021
  • In this paper we define scaled visual curvature and visual Frenet frame that can be visually accepted for discrete space curves. Scaled visual curvature is relatively simple compared to multi-scale visual curvature and easy to control the influence of noise. We adopt scaled minimizing directions of height functions on each neighborhood. Minimizing direction at a point of a curve is a direction that makes the point a local minimum. Minimizing direction can be given by a small noise around the point. To reduce this kind of influence of noise we exmine the direction whether it makes the point minimum in a neighborhood of some size. If this happens we call the direction scaled minimizing direction of C at p ∈ C in a neighborhood Br(p). Normal vector of a space curve is a second derivative of the curve but we characterize the normal vector of a curve by an integration of minimizing directions. Since integration is more robust to noise, we can find more robust definition of discrete normal vector, visual normal vector. On the other hand, the set of minimizing directions span the normal plane in the case of smooth curve. So we can find the tangent vector from minimizing directions. This lead to the definition of visual tangent vector which is orthogonal to the visual normal vector. By the cross product of visual tangent vector and visual normal vector, we can define visual binormal vector and form a Frenet frame. We examine these concepts to some discrete curve with noise and can see that the scaled visual curvature and visual Frenet frame approximate the original geometric invariants.

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

  • 윤성식;김창민
    • Microbiology and Biotechnology Letters
    • /
    • v.29 no.1
    • /
    • pp.1-11
    • /
    • 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.

  • PDF

Construction of Indoor and Outdoor Spatial Information Integration Service System based on Vector Model

  • Kim, Jun Hyun;Kwon, Kee Wook
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
    • /
    • v.36 no.3
    • /
    • pp.185-196
    • /
    • 2018
  • In order to overcome the problem that outdoor and indoor spatial information service are separately utilized, an integration service system of spatial information that is linked from outdoor to indoor has been implemented. As a result of the study, "0001.xml" corresponding to the file index key value, which is the service connection information in the building information of the destination, was extracted from the prototype verification of the system, the search word of 'Kim AB' was transmitted to the indoor map server and converted from the outdoor map service to the indoor map service through confirmation of the navigation service connected information, using service linkage information and search words of the indoor map service was confirmed that the route was displayed from the entrance of the building to the destination in the building through the linkage search DB (Database) table and the search query. Therefore, through this study was examined the possibility of linking indoor and outdoor DB through vector spatial information integration service system. The indoor map and the map engine were implemented based on the same vector map format as the outdoor map engine, it was confirmed that the connectivity of the map engine can be applied.

A Study on the Application Technique and Integration of Remote Sensing and Geographic Information System (리모트센싱과 GIS의 통합 및 그 적용기법에 관한 연구)

  • 안철호;연상호
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
    • /
    • v.9 no.1
    • /
    • pp.97-107
    • /
    • 1991
  • This paper was suggested the detailed methods on the integration of Remote Sensing and GIS for various application of two functions at the one system with making the most use of respective merits rather than make use of independent systems. It developed of algorithm about simultaneous overlay of raster and vector data for remote sensing and GIS for these objects. For test application on integration of remote sensing and GIS, it used of remote sensing data of satellite and used to topographic map of the same area for vector data acquisition of GIS application. For the practical application, it proved of effective value of integration of raster and vector data by present of useful technique with multilateral approach method through data conversion about thematic application for major application fields of remote sensing and GIS and it suggested that new application technique for integrated application of remote sensing GIS through synthetic situation analysis.

  • PDF

A Novel Integrative Expression Vector for Sulfolobus Species

  • Choi, Kyoung-Hwa;Hwang, Sungmin;Yoon, Naeun;Cha, Jaeho
    • Journal of Microbiology and Biotechnology
    • /
    • v.24 no.11
    • /
    • pp.1503-1509
    • /
    • 2014
  • With the purpose of facilitating the process of stable strain generation, a shuttle vector for integration of genes via a double recombination event into two ectopic sites on the Sulfolobus acidocaldarius chromosome was constructed. The novel chromosomal integration and expression vector pINEX contains a pyrE gene from S. solfataricus P2 ($pyrE_{sso}$) as an auxotrophic selection marker, a multiple cloning site with histidine tag, the internal sequences of malE and malG for homologous recombination, and the entire region of pGEM-T vector, except for the multiple cloning region, for propagation in E. coli. For stable expression of the target gene, an ${\alpha}$-glucosidase-producing strain of S. acidocaldarius was generated employing this vector. The malA gene (saci_1160) encoding an ${\alpha}$-glucosidase from S. acidocaldarius fused with the glutamate dehydrogenase ($gdhA_{saci}$) promoter and leader sequence was ligated to pINEX to generate pINEX_malA. Using the "pop-in" and "pop-out" method, the malA gene was inserted into the genome of MR31 and correct insertion was verified by colony PCR and sequencing. This strain was grown in YT medium without uracil and purified by His-tag affinity chromatography. The ${\alpha}$-glucosidase activity was confirmed by the hydrolysis of $pNP{\alpha}G$. The pINEX vector should be applicable in delineating gene functions in this organism.