• Title/Summary/Keyword: DNA-DNA hybridization

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Improvement of Glucoamylase Productivity of Saccharomyces diastaticus by Intergration of Glucoamylase Gene, STA, into Chromosomal DHA (Glucoamylase 유전자 STA의 염색체내 삽입에 의한 Saccharomyces diastaticus의 glucoamylase 생성능 향상)

  • 안종석;맹준호;강대욱;황인규;민태익
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.48-53
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    • 1993
  • For the purpose to improve the glucoamylase productivity of Saccharomyces diastaticus, we integrated STA 1 gene into chromosomal DNA of S. diastaticus using YIp vector. After construction of Ylp-STA by the subcloning of STAI (5.3 kb) into YIp5 vector, S. diastaticus GMT-II(a. ura3. STAJ) was transformed by Ylp-STA through homologous recombination at the chromosomal STAJ gene. So we obtained the tram formants that glucoamylase productivity was increased maximum six fold. These strains transformed by the multi-copy integration of Ylp-STA in chromosomal DNA were confirmed by Southern hybridization. And the integrated Ylp-STA was maintained stably during 30 mitotic divisions.

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Automatic Reading System for On-off Type DNA Chip

  • Ryu, Mun-Ho;Kim, Jong-Dae;Kim, Jong-Won
    • Journal of Information Processing Systems
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    • v.2 no.3 s.4
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    • pp.189-193
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    • 2006
  • In this study we propose an automatic reading system for diagnostic DNA chips. We define a general specification for an automatic reading system and propose a possible implementation method. The proposed system performs the whole reading process automatically without any user intervention, covering image acquisition, image analysis, and report generation. We applied the system for the automatic report generation of a commercialized DNA chip for cervical cancer detection. The fluorescence image of the hybridization result was acquired with a $GenePix^{TM}$ scanner using its library running in HTML pages. The processing of the acquired image and the report generation were executed by a component object module programmed with Microsoft Visual C++ 6.0. To generate the report document, we made an HWP 2002 document template with marker strings that were supposed to be searched and replaced with the corresponding information such as patient information and diagnosis results. The proposed system generates the report document by reading the template and changing the marker strings with the resultant contents. The system is expected to facilitate the usage of a diagnostic DNA chip for mass screening by the automation of a conventional manual reading process, shortening its processing time, and quantifying the reading criteria.

Identification of Biomarkers for Radiation Response Using cDNA Microarray

  • Park, Woong-Yang
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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    • pp.29-44
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    • 2001
  • DNA damage by physical insult including UV and g-radiation might provoke genetic alterations in cells, which is followed by either acute cell death or tumorigenesis. The responsiveness to g-radiation depends on cellular context of target cells. To understand the mechanisms of checkpoint control, repair and cell death following genotoxic stimu]i, cDNA microarray can provide the gene expression profile. To make a profile of gene expression in irradiated Jurkat T cells, we hybridized the cDNA microarray using cDNA from g-irradiated Jurkat T cells. Jurkat T cells were exposed to 4Gy to 16Gy, and total RNA were extracted at 4 to 24 hrs after irradiation. The hybridization of the microarray to fluorescence-labeled cDNA from treated and untreated cells was analyzed by bioinformatic analysis to address relative changes in expression levels of the genes present in the array. Responses varied widely in different time points, suggesting acute stress response and chronic restoration or cell death. From these results we could select 384 genes related to radiation response in Tcells, and radiation response might be different in various types of cells. Using Radchip, we could separate "the exposed" from control PBMCs. We propose that Radchip might be useful to check the radiation research as well as radiation carcinogenesis.

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Prognosis of Colorectal Cancer and Apoptosis Activation (대장암의 예후와 Apoptosis 활성)

  • 전소영;배옥석;김종봉
    • Biomedical Science Letters
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    • v.4 no.2
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    • pp.121-128
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    • 1998
  • We studied on the expression of apoptosis in colorectal cancer, lymph node, their corresponding normal mucosa and colorectal cancer patient's blood by genomic DNA electrophoresis and TUNEL labeling method. From 7 cases among 37, 20 cases among 47 and 5 cases among 15, DNA ladders were expressed in normal tissues, colorectal tissues and Iymph node tissues, respectively. A DNA ladder was not observed in 7 cases of colorectal cancer patients blood. In case of TUNEL labeling, we could observe TUNEL color espression in colorectal cancer and lymph node tissues. As these result suggest that apoptotic index may be associated with the colorectal cancer development, and mat be used as a prognostic indicator but further evaluations will be needed.

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Cloning of the rbcL Gene from Maize Chloroplast (옥수수 엽록체 rbcL 유전자의 클로닝)

  • 이재선
    • Journal of Plant Biology
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    • v.35 no.2
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    • pp.165-171
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    • 1992
  • In order to study regulation of rbcL gene expression, rbcL gene of chloroplast DNA (Cp DNA) from maize was cloned. Cp DNA was isolated from intact chloroplast and digested with BamHI. BamHI 9 fragment of Cp DNA containing rbcL gene was ligated to pUC19 and transformed into E. coli DH5a. This recombinant plasmid was named pRLYSl. pRLYSl was hybridized with a part of rbcL gene from rice and digested with restriction enzyme BamHI, HindIIl, and PstI. From these results, it was confirmed that pRLYS1 contains intact rbcL gene and orientation of BamHI 9 fragment of Cp DNA in pRLYS1 was determined.rmined.

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Specific Detection of DNA Using Quantum Dots and Magnetic Beads for Large Volume Samples

  • Kim, Yeon-Seok;Kim, Byoung-Chan;Lee, Jin-Hyung;Kim, Jung-Bae;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.449-454
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    • 2006
  • Here we present a sensitive DNA detection protocol using quantum dots (QDs) and magnetic beads (MBs) for large volume samples. In this study, QDs, conjugated with streptavidin, were used to produce fluorescent signals while magnetic beads (MBs) were used to isolate and concentrate the signals. The presence of target DNAs leads to the sandwich hybridization between the functionalized QDs, the target DNAs and the MBs. In fact, the QDs-MBs complex, which is bound using the target DNA, can be isolated and then concentrated. The binding of the QDs to the surface of the MBs was confirmed by confocal microscopy and Cd elemental analysis. It was found that the fluorescent intensity was proportional to concentration of the target DNA, while the presence of non-complementary DNA produced no significant fluorescent signal. In addition, the presence of low copies of target DNAs such as 0.5 pM in large volume samples up to 40mL was successfully detected by using a magnet-assisted concentration protocol which consequently results in the enhancement of the sensitivity more than 100-fold.

Transformation of Coprinus congregatus with a Linearized Plasmid Vector to Phosphinothricin Resistance (Coprinus congregatus에서 선형으로 전환한 plasmid DNA를 사용하여 phosphinothricin 저항성에 대한 형질전환)

  • Leem, Young-Eun;Kim, Soon-ja;Choi, Hyoung-Tae
    • Korean Journal of Microbiology
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    • v.33 no.4
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    • pp.274-276
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    • 1997
  • Transformation of Coprinus congregatus with a linearized plasmid has been successfully carried out using phosphinothricin resistance gene as a dominant selectable marker. The transforming frequency was about 500 transformants per microgram of DNA using the protoplast-$CaCl_2$ method. The transforming vector pBARGEM 7-1 which had the phosphinothricin resistance gene was detected in the restriction enzyme fragments of chromosomal DNA from a transformant by Southern hybridization.

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Genetic diversity and phylogenetic analysis of genus Paeonia based on nuclear ribosomal DNA ITS sequence

  • Sun, Yan-Lin;Hong, Soon-Kwan
    • Journal of Plant Biotechnology
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    • v.38 no.3
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    • pp.234-240
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    • 2011
  • The genus Paeonia belongs to the family Paeoniaceae having significant medicinal and ornamental importance. The present investigation was undertaken with an aim to understand phylogenetic relationships of three Paeonia species (P. lactiflora, P. obovata, and P. suffruticosa) that are widely distributed in China, Korea, and Japan, using nuclear ribosomal DNA (nrDNA) internal transcribed spacer (ITS) sequence and to compare the phylogeny results with investigations reported earlier using existed sequences of the same species. The size variation obtained among sequenced nrDNA ITS region was narrow and ranged from 722 to 726 bp. The highest interspecific genetic distance (GD) was found between P. lactiflora and P. suffruticosa or P. obovata. The phylogram obtained using our nrDNA ITS sequences showed non-congruence with previous hypothesis of the phylogeny between section Paeonia and section Moutan of genus Paeonia. This result was supported by the phylogenetic relations showed in the phylogram constructed with existed sequences in NCBI. The present study suggested that P. obovata belonging to section Paeonia was phylogenetically closer to P. suffruticosa representing section Moutan of genus Paeonia than P. lactiflora belonging to section Paeonia. The main reason of the paraphyly of section Paeonia is thought to be nucleotide additivity directly caused by origin hybridization. This study provides more sequence sources of genus Paeonia, and will help for further studies in intraspecies population, and their phylogentic analysis and molecular evolution.

A Study on the Molecular Biology of Renin-Angiotensin System : Renin Gene Expression and Construction of Genomic Library (Renin-Angiotensin계의 분자생물학적 연구 : Renin유전자의 발현과 Genomic Library작성)

  • 박영순;한동민;김종호;문영희;이호섭;고건일;김성준
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.35-44
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    • 1990
  • Poly (A) + RNA was isolated from mouse submaxillary gland and renin mRNA was isolated by poly (U)-sepharose chromatography and sucrose linear densiW gradient centifugation. And renin mRNA was identified by in vitro translation and immunoprecipitation. In order to construct recombinant plasmid, renin cDNA was synthesized by using reverse transcriptase and inserted into EcoRi site of PUC19. In addition, the cDNA was also synthesized using polymerase chain reaction and inserted into HindlIl site of PUC19. The recombinant plasmid was transformed into JMlO3 and the expression of the inserted renin cDNA was examined. The transformant produced renin protein having a molecular weight of 45, 000 dolton, which showed hypertensive effect upon injecting it into rabbit ear vein. A renin genomic library was prepared by inserting rabbit kidney DNA into EMBL3 phage, and was screeined for the isolation of renin gemomic DNA using renin cDNA probe.

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Isolation of Differentially Expressed Genes by Low Temperature Treatment in Winter Oilseed Rape (Brassica napus L. cv. Tammi)

  • Chun Jong-Un;Seo Dong-Joong;Bae Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.19 no.3
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    • pp.440-446
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    • 2006
  • To investigate flowering related genes in winter-type oilseed rape (Brassica napus L. cv. Tammi), differentially expressed genes were isolated from leaves of the plant after low temperature treatment which is requirements for floral induction. As a result of suppression subtractive hybridization (SSH), 288 clones were randomly selected from SSH library. Using reverse Northern blot analysis, 150 of 288 clones were identified to be differentially expressed. Out of these 150 clones, 45 clones showed very high identities with the known genes. Four clones showed very high identities over 90% with metallothionein-like gene that is related to flowering-induced genes. Of these 4 clones, the cDNA clone, rfs-13, revealed high identity with meotallothionein-like protein in Arabidopsis thaliana (98%) and Brassica compestris (89%). Furthermore, gene expressed in immature flower stages was confirmed by Northern blot analysis.