• Title/Summary/Keyword: Cloning detection

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Preservation through Cloning of Superior Canine Scent Detection Ability for Cancer Screening (복제를 통한 우수한 암탐지 능력의 보존)

  • Kim, Min-Jung;Park, Jung-Eun;Oh, Hyun-Ju;Hong, So-Gun;Kang, Jung-Taek;Rhim, Sang-Hyun;Lee, Dong-Won;Ra, Jung-Chan;Lee, yeong-Chun
    • Journal of Veterinary Clinics
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    • v.32 no.4
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    • pp.352-355
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    • 2015
  • This study was conducted to ascertain whether the scent detection ability of a donor dog having extraordinary talent in cancer detection can be conserved through cloning. A specially trained dog for colorectal cancer detection was cloned, and she was trained and tested to detect breast cancers using breath samples collected from patients and healthy volunteers. Scent detection sensitivity of the clone was 93.3% and specificity was 99.5%, similar with those of donor (91% and 99%). Furthermore, the clone successfully detected early stage of breast cancers. Therefore, superior canine scent detection ability for cancer screening could be preserved through cloning.

Block and Fuzzy Techniques Based Forensic Tool for Detection and Classification of Image Forgery

  • Hashmi, Mohammad Farukh;Keskar, Avinash G.
    • Journal of Electrical Engineering and Technology
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    • v.10 no.4
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    • pp.1886-1898
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    • 2015
  • In today’s era of advanced technological developments, the threats to the authenticity and integrity of digital images, in a nutshell, the threats to the Image Forensics Research communities have also increased proportionately. This happened as even for the ‘non-expert’ forgers, the availability of image processing tools has become a cakewalk. This image forgery poses a great problem for judicial authorities in any context of trade and commerce. Block matching based image cloning detection system is widely researched over the last 2-3 decades but this was discouraged by higher computational complexity and more time requirement at the algorithm level. Thus, for reducing time need, various dimension reduction techniques have been employed. Since a single technique cannot cope up with all the transformations like addition of noise, blurring, intensity variation, etc. we employ multiple techniques to a single image. In this paper, we have used Fuzzy logic approach for decision making and getting a global response of all the techniques, since their individual outputs depend on various parameters. Experimental results have given enthusiastic elicitations as regards various transformations to the digital image. Hence this paper proposes Fuzzy based cloning detection and classification system. Experimental results have shown that our detection system achieves classification accuracy of 94.12%. Detection accuracy (DAR) while in case of 81×81 sized copied portion the maximum accuracy achieved is 99.17% as regards subjection to transformations like Blurring, Intensity Variation and Gaussian Noise Addition.

A Modified Mutation Detection Method for Large-scale Cloning of the Possible Single Nucleotide Polymorphism Sequences

  • Jiang, Ming-Chung;Jiang, Pao-Chu;Liao, Ching-Fong;Lee, Ching-Chiu
    • BMB Reports
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    • v.38 no.2
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    • pp.191-197
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    • 2005
  • Although the human genome has been nearly completely sequenced, the functions and the roles of the vast majority of the genes, and the influences of single nucleotide polymorphisms (SNPs) in these genes are not entirely known. A modified mutation detection method was developed for large-scale cloning of the possible SNPs between tumor and normal cells for facilitating the identification of genetic factors that associated with cancer formation and progression. The method involves hybridization of restriction enzyme-cut chromosomal DNA, cleavage and modification of the sites of differences by enzymes, and differential cloning of sequence variations with a designed vector. Experimental validations of the presence and location of sequence variations in the isolated clones by PCR and DNA sequencing support the capability of this method in identifying sequence differences between tumor cells and normal cells.

Construction of Bioluminescent Escherichia coli from lux Operon and Heat Shock Promoter for the Detection of Toxic Substances (lux Operon과 Heat Shock Promoter 유전자 재조합을 통한 독성물질 탐지용 대장균의 개발)

  • 유승오;이은관;김현숙;정계훈;전억한
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.278-285
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    • 1999
  • In order to use heat shock promoter for the detection of toxic substances, dnaK promoter was amplified from E. coli genomic DNA by using a polymerase chain reaction(PCR) followed by sequencing and sub-cloning into the multi-cloning site of the plasmid, pUCD615. The pUCD615 is a broad-host-range vector containing promoterless lux operon originated from V.fischeri. The recombinant plasmid was transfered to E. coli DH5$\alpha$ through electroporation. The recombinant E. coli showed several patterns of bioluminescent responses to ethanol stress. The bioluminescent E. coli also showed responses to other toxic substances including FeK3(CN)6, CdCl2, p-nitrophenol and HgCl2. The increases of RLU(Relative Light Unit) were observed at 100ppm of FeK3(CN)6, 10ppm and 100ppm and 100ppm of CdCl2, 1ppm of 10ppm of p-nitrophenol and at 1ppm of HgCl2.

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Molecular Cloning of the Gene for $\alpha$-Acylamino-$\beta$-lactam Acylhydrolase from Acetobacter turbidans by Immunochemical Detection Method (면역화학적 방법에 의한 Acetobacter turbidans의 $\alpha$-Acylamino-$\beta$-lactam Acylhydrolase의 유전자 클론화)

  • Nam, Doo-Hyun;Dewey D.Y. Ryu
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.363-368
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    • 1988
  • Molecular cloning of gene for $\alpha$-acylamino-$\beta$-lactam acylhydrolase (ALAH) III from Acetobacter turbidans has been attempted by immunochemical detection method, in which polyclonal antibody from mouse Balb/c against this enzyme was employed as a probe. As a cloning vector, λ gtll was chosen for this purpose. Two positive clones has been selected from genomic libraries of A. turbidans, which had somewhat different binding affinities on anti-ALAH III umm and anti-$\beta$-galactosidase. By restriction analysis, both clones has been turned out to lose one of EeoRI sites. From these results, it concluded that deletion of DNA between lacZ gene and inserted DNA has occurred during replication of these clones in host cells.

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A study on the Aptamer Specific Detection on P. gingivalis (P. gingivalis에 특이적으로 작용하는 앱타머에 관한 연구)

  • Shin, Ae-Ri
    • The Journal of the Korea Contents Association
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    • v.21 no.4
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    • pp.825-832
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    • 2021
  • In this study, by selecting specific aptamers that selectively detection on P. gingivalis, the main cause of periodontal disease, and purifying and identifying protein molecules that bind to the selected aptamers, the mechanism of action of P. gingivalis was investigated. A DNA library having 39 random sequences was prepared, and aptamers with specificity for P. gingivalis were selected using the SELEX method, and the nucleotide sequence was analyzed by cloning using PCR2.1 cloning vector. 8 of aptamers with different nucleotide sequences were selected, and modified weston blot was performed using APG-3 among the selected aptamers to identify 11 proteins that act directly, and proteins were analyzed. As a result, a protein that selectively binds to P. gingivalis was isolated and identified. Therefore, aptamer selectively binds and attaches to proteins related to inhibition of sugar metabolism and cell activity of P. gingivalis, suggesting the possibility of a sensor for diagnosis of periodontal disease.

Molecular Cloning and Nucleotide Sequence of Xylanase gene (xynT) from Bacillus alcalophilus AX2000. (Bacillus alcalophilus AX2000 유래 xylanase 유전자 (XynT)의 Cloning과 염기서열 분석)

  • Park Young-Seo
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.734-738
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    • 2005
  • A gene coding for xylanase from alkali-tolerant Bacillus alcalophilus AX2000 was cloned into Escherichia coli $DH5\alpha$ using pUC19. Among 2,000 transformants, one transformant showed clear zone on the detection agar plate containing oat-spells xylan. Its recombinant plasmid, named pXTY99, was found to carry 7.0 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynT) was determined, xynT gene was found to consist of 1,020 base-pair open reading frame coding for a poly-peptide of 340 amino acids with a deduced molecular weight of 40 kDa. The coding sequence was preceded by a putative ribosome binding site, and the transcription initiation signals. The deduced amino acid sequence of xylanase is similar to those of the xylanases from Bacillus sp. Nl37 and B. stearothermophilus 21 with $61\%$ and $59\%$ identical residues, respectively.

Molecular Detection of Harmful Dinoflagellates (Dinophyceae) in Ballast Water (선박평형 수 내 유해 와편모조류(Dinophyceae)의 분자생물학적 검출)

  • Park, Tae-Gyu;Kim, Sung-Yeon
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.1
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    • pp.36-40
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    • 2010
  • Ballast water has been known as a major vector for global dispersal of toxic dinoflagellates and other microalgae. In this study, biodiversity in ships’ ballast water was examined using a dinoflagellate-oriented PCR primer set and species-specific real-time PCR. While motile dinoflagellates could be observe at very low cell densities by light microscopy,a wide range of dinoflagellate taxa including parasitic and phototrophic pico-dinoflagellates as well as harmful species to marine fish/shellfish was detected when techniques for cloning/sequencing of SSU rDNA of sample cells were used. Present result suggests that molecular methods including species-specific PCR primers may offer rapid and accurate detection of invasive species in ballast water.

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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