• 제목/요약/키워드: 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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    • 제11권3호
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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.

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

  • 임향숙;최주연;김인경;강성만;성영모
    • 미생물학회지
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    • 제35권1호
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    • pp.28-34
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    • 1999
  • 본 연구에서는 Yeast Artificial Chromosome을 효율적으로 조작하고 유유전자의 기능을 보다 더 용이하게 분석하기 위해 EMCV 바이러스의 IRES 염기서열과 $\beta$-galactosidase를 포함하는 새로운 bicistronic fragmentation vector를 제조하였다. 이 벡터의 폴리클로닝 site에 네 개의희귀한 제한효소 부위를 도입하여 DNA를 용이하게 클로닝할 수 있게 만들었다. 그러므로 원하는 어떤 YAC도 효모세포에서 쉽게 상동 재조함에 의해 절편할 수 있는 장점이 있다. 이 bicistronic fragmentation vector 시스템을 이용하면하나의 메시지로부터 연구하고자 하는 유전자와 $\beta$-galactosidase를 동시에 한 세포에서 발현할 수 있기 때문에 유전자의 발현양상을 쉽게 분석할 수 있는 새로운 도구로 이용할 수 있다.

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

  • 조무제;신평균;최영주;강규영;윤한대
    • 미생물학회지
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    • 제27권2호
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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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체세포 융합에 의한 닭의 유전인자구명에 관한 연구 (Identification of Gene Locus by the Somatic Cell Hybridization in Chicken)

  • 정익정
    • 한국가금학회지
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    • 제16권1호
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    • pp.1-8
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    • 1989
  • 유전물질의 총합체인 염색체의 분리와 분석을 정확히 하여 유전인자의 위치를 밝히고 또한 유전자조작 기술을 이용할 수 있는 방법을 구명하여 닭의 능력개양을 꾀하기 위해서 실시된 본 연구의 결과를 요약하면 아래와 같다. 1. 백혈구 배양을 통하며 염색체를 분리하고 Giemsa banding을 통한 염색체 분석에서 1번 염색체의 경우 20층에 달하는 banding pattern을 발견할 수 있는 정확한 분석으로 1∼9번, 성염색체의 정상 banding pattern을 밝힐 수 있었고 C-banding의 결과 모든 염색체에서 유전자 작용이 없는 constitutive heterochromatin의 위치를 밝힐 수 있었다. 2. 유전자 조작 기술 중 중요한 단계인 genetic vector로서 닭의 원시생식세포(Primodial germ cell, PGC)를 이용하기 위하여 genetic marker로서 3배체의 염색체를 가친 PGC를 정상 host embryo에 이식시켜서 성장중인 host embryo의 성선에서 donor PGC의 genetic marker(3n)가 발견됨으로써 PGC를 이용한 가금의 유전자 조작이 가능함을 밝혔다.

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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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    • 제9권6호
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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.

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

  • 윤성식;김창민
    • 한국미생물·생명공학회지
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    • 제29권1호
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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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인터넷을 통한 벡터 공간 데이타의 효율적 전송을 위한 최적화 기법 (An Optimization Strategy for Vector Spatial Data Transmission onover the Internet)

  • Liang Chen;Chung-Ho Lee;Hae-Young Bae
    • 한국정보과학회논문지:데이타베이스
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    • 제30권3호
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    • pp.273-285
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    • 2003
  • 일반적으로 공간 벡터 데이타는 래스터 데이타에 비해 많은 정보를 포함하고 있으므로, 좀 더 융통적이고 효율적으로 데이타에 대한 처리가 가능하다. 그러나 인터넷을 통한 공간 벡터 데이타의 조작 시 해결해야 할 문제로 좁은 대역폭을 갖는 인터넷에서 크기가 크고 복잡한 벡터 데이타를 어떻게 효율적으로 전송하는가 라는 문제이다. 본 논문은 좁은 대역폭을 갖는 인터넷을 통한 공간 벡터 데이타를 효율적으로 전송하기 위한 새로운 전송 기법인 스케일에 기반한 전송 기법을 제안한다. 제안된 기법의 아이디어는 보여질수 있는 것만을 전송하는 것이다. 특정 스케일에서 일부 피쳐만이 사용자에게 보여지므로, 자연히 스케일은 공간 피쳐와 연관된 요소이다. 제안된 기법은 웨이블릿에 기반한 지도 일반화 알고리즘을 통해 공간 객체 중에서 출력되는 스케일에 따라 보여질 필요가 없는 피쳐들을 필터링하고, 보여지는 피쳐만을 최종적으로 전송한다. 본 논문에서는 실험을 통해 제안된 기법을 사용하는 경우, 개개의 공간 연산들에 대한 응답 시간이 대체적으로 향상됨을 보인다.

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

  • 정지아;이하나;고인정;오정일
    • 생명과학회지
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    • 제23권2호
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    • pp.290-298
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    • 2013
  • Mycobacterium 속은 Mycobacterium tuberculosis, Mycobacterium leprae, Mycobacterium bovis와 같은 동물과 인체에 병원성을 나타내는 세균 종을 다수 포함하고 있다. 이들의 숙주에서의 생존과 병원성에 관한 유전학적 정보를 확보하는 것은 매우 중요하지만, 효과적인 유전학적 도구가 부족하였기 때문에 이들에 관한 연구가 미비하였다. 따라서 mycobacteria의 연구를 위한 분자생물학적 실험 도구로서 다양한 기능성 vector들이 고안되었고, 이러한 기능성 vector의 개발은 실질적으로 mycobacteria에서의 연구 효과를 증진시켰다. 본 연구에서는 Mycobacterium smegmatis에 적용 가능하고 기존에 제시되었던 mycobacteria 연구에 있어서의 한계점을 극복하기 위한 노력의 일환으로, 기능성 vector인 temperature-sensitive replication origin (TSRO)과 counterselectable marker로 levansucrase를 암호화하는 sacB 유전자를 포함하는 suicide vector pKOTs, chromosomal DNA로 site-specific recombination을 통해 삽입되는 lacZ transcriptional fusion vector pMV306lacZ, 그리고 TSRO를 가지는 minitransposon vector pTnMod-OKmTs를 개발하였다. 이 vector들은 실질적으로 M. smegmatis에서 효과적으로 작동하는 것이 확인되었으며 목적으로 하는 실험 결과 도출 가능성 또한 보여주었다. 따라서 이들 vector는 앞으로의 mycobacteria에 대한 효과적인 연구 기반이 될 것으로 기대된다.

Manipulation of Hepatitis B Viral DNA for Generating Transgenic Mice

  • Kim, Seung-Hee;Park, Sang-Ho;Kim, Tae-Gyun;Lee, Song-Deuk;Aree Moon
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1996년도 춘계학술대회
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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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    • 제32권5호
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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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