• Title/Summary/Keyword: Gene Gun

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Correlation Analysis between Regulatory Sequence Motifs and Expression Profiles by Kernel CCA

  • Rhee, Je-Keun;Joung, Je-Gun;Chang, Jeong-Ho;Zhang, Byoung-Tak
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.63-68
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    • 2005
  • Transcription factors regulate gene expression by binding to gene upstream region. Each transcription factor has the specific binding site in promoter region. So the analysis of gene upstream sequence is necessary for understanding regulatory mechanism of genes, under a plausible idea that assumption that DNA sequence motif profiles are closely related to gene expression behaviors of the corresponding genes. Here, we present an effective approach to the analysis of the relation between gene expression profiles and gene upstream sequences on the basis of kernel canonical correlation analysis (kernel CCA). Kernel CCA is a useful method for finding relationships underlying between two different data sets. In the application to a yeast cell cycle data set, it is shown that gene upstream sequence profile is closely related to gene expression patterns in terms of canonical correlation scores. By the further analysis of the contributing values or weights of sequence motifs in the construction of a pair of sequence motif profiles and expression profiles, we show that the proposed method can identify significant DNA sequence motifs involved with some specific gene expression patterns, including some well known motifs and those putative, in the process of the yeast cell cycle.

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Structural Analysis and Transcriptional Regulation of the Chloroplast psbC Gene from Panax ginseng

  • Yoo, Ki-Yeol;Tae, Gun-Sik
    • Journal of Photoscience
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    • v.12 no.3
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    • pp.129-133
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    • 2005
  • The psbC gene, encoding the intrinsic chlorophyll-binding protein of CP43, one of the PS core complex polypeptides, was cloned from the Panax ginseng chloroplast, which is composed of 1,422 nucleotides and the overall nucleotide sequence shows more than 84% identity to those of eukaryotic photosynthetic organisms. The predicted topology of CP43, based on hydropathy analysis, includes six membrane-spanning ${\alpha}-helices$ resulting in three lumenal and four stromal loops. The putative translation start codon for the psbC gene is located at 48 nucleotides upstream from the stop codon of the psbD gene whose product is also a component of the PSII reaction center, implying that the promoter of the psbC gene is possibly located in the middle of the structural gene of the psbD gene. Northern blot analysis of the in vivo accumulation of the psbC transcript from the plants grown under the various growth light intensities (5%, 10%, 20%, and 100%) of daylight indicated that the steady-state level of the psbC transcript was not significantly affected by light intensity.

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Egr-1 regulates the transcription of the BRCA1 gene by etoposide

  • Shin, Soon Young;Kim, Chang Gun;Lee, Young Han
    • BMB Reports
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    • v.46 no.2
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    • pp.92-96
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    • 2013
  • The breast cancer susceptibility gene BRCA1 encodes a nuclear protein, which functions as a tumor suppressor and is involved in gene transcription and DNA repair processes. Many families with inherited breast and ovarian cancers have mutations in the BRCA1 gene. However, only a few studies have reported on the mechanism underlying the regulation of BRCA1 expression in humans. In this study, we investigated the transcriptional regulation of BRCA1 in HeLa cells treated with etoposide. We found that three Egr-1-binding sequences (EBSs) were located at -1031, -1005, and -385 within the enhancer region of the BRCA1 gene. Forced expression of Egr-1 stimulated the BRCA1 promoter activity. EMSA data showed that Egr-1 bound directly to the EBS within the BRCA1 gene. Knockdown of Egr-1 through the expression of a small hairpin RNA (shRNA) attenuated etoposide-induced BRCA1 promoter activity. We conclude that Egr-1 targets the BRCA1 gene in HeLa cells exposed to etoposide.

Molecular Cloning of Bacillus stearothermophilus cdd Gene Encoding Thermostable Cytidine/Deoxycytidine Deaminase (Bacillus stearothermophilus 의 내열성 시티딘/디옥시시티딘 디아미나제를 코드하는 cdd 유전자의 클로닝)

  • Soo, Chang-Jong;Song, Bang-Ho;Kim, Jong-Guk;Hong, Soon-Duck
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.334-342
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    • 1989
  • The Bacillus stearothermophilus cdd gene encoding cytidine deaminase (cytidine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5) was isolated through shot gun cloning by oomplementation of an E. coli cdd mutation. Primarily 3.0 kbp of the exogenote was cloned into the Pstl site of pBR322 (pJSC101). By subsequent deletion and subcloning from the insert of pJSC101 with cdd$^+$ and tetracycline resistancy, about 1.35 kbp of the EcoRI$_1$/PstI$_2$ fragment containing the cdd gene was isolated as pJSC201. The minicell experiment revealed a molecular mass of 33,000 dalton for polypeptide from the cloned DNA fragment complementing the cdd gene. From the lacZ fusion of 550 bp fragment of the EcoRI$_1$/AuaI as a putative promoter region, the transcription direction of the cdd gene on pJSC201 is from EcoRI towards the PstI sites, When the cdd gene was expressed in B. subtilis ED4O (cdd$^-$, pyr$^-$) by transformation with the E. coli-B. subtilis shuttle vector, the gene expression occured more efficiently than in E. coli and the gene appears to be stably maintained in B. subtitis as well as in E. coli.

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Gene Sequences Clustering for the Prediction of Functional Domain (기능 도메인 예측을 위한 유전자 서열 클러스터링)

  • Han Sang-Il;Lee Sung-Gun;Hou Bo-Kyeng;Byun Yoon-Sup;Hwang Kyu-Suk
    • Journal of Institute of Control, Robotics and Systems
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    • v.12 no.10
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    • pp.1044-1049
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    • 2006
  • Multiple sequence alignment is a method to compare two or more DNA or protein sequences. Most of multiple sequence alignment tools rely on pairwise alignment and Smith-Waterman algorithm to generate an alignment hierarchy. Therefore, in the existing multiple alignment method as the number of sequences increases, the runtime increases exponentially. In order to remedy this problem, we adopted a parallel processing suffix tree algorithm that is able to search for common subsequences at one time without pairwise alignment. Also, the cross-matching subsequences triggering inexact-matching among the searched common subsequences might be produced. So, the cross-matching masking process was suggested in this paper. To identify the function of the clusters generated by suffix tree clustering, BLAST and CDD (Conserved Domain Database)search were combined with a clustering tool. Our clustering and annotating tool consists of constructing suffix tree, overlapping common subsequences, clustering gene sequences and annotating gene clusters by BLAST and CDD search. The system was successfully evaluated with 36 gene sequences in the pentose phosphate pathway, clustering 10 clusters, finding out representative common subsequences, and finally identifying functional domains by searching CDD database.

DD1.5k, the Gene Preferentially Expressed in Bloodstream Isolates of Vancomycin-Resistant Enterococcus faecium

  • Kim, Seung-Han;Lee, Dong-Gun;Yoo, Jin-Hong;Park, Su-Mi;Park, Jung-Hyun;Shin, Wan-Shik;Lee, Kyungwon;Dongeun Yong;Lee, Wee-Gyo
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.143-146
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    • 2004
  • Vancomycin-resistant Enterococcus faecium (VREFM) is becoming a threatening pathogen. We identified a gene called DD1.5K by differential display-PCR, which was preferentially expressed in the bloodstream isolates of VREFM. Due to its amino acid similarity to transfer complex protein, trsE, and tissue-specific expression, this gene may be involved in virulence of VREFM.

Gene Cloning of Cellulose Degradation Enzyme of Bacillus subtilis LYH201 Strain (Bacillus subtilis LYH201균주의 섬유소 분해효소의 유전자 Cloning 및 특성분석)

  • Lee, Young-Han;Park, Sang-Ryeol
    • Korean Journal of Soil Science and Fertilizer
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    • v.34 no.5
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    • pp.333-341
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    • 2001
  • The Compost-decomposing-bacteria was isolated from livestock compost containing sawdust. The isolated bacteria was identified as Bacillus subtilis LYH201 by the method of the composition of the fatty acid with MIDI system and Bergey's manual. Cloning of CMCase encoding gene was accompanied by shotgun method. The pLK100 have yellow activity ring on CMC medium, that was carried 2.2 kb insert DNA in pBluescript II $SK^+$ vector, named BglC gene. The BglC was very similar to Pectobacterium carotovorum Gun_CLOAB(P15704) with score of 57% identity and 71% homology over 508 aa. The BglC was measured molecular weight 56 kDa by CMC-SDS-PAGE. Optimum cellulase activity Bacillus subtilis LYH201 was temperature $50^{\circ}C$ and pH 7.

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Cloning and Expression of a bpr Gene Encoding Bacillopeptidase F from Bacillus amyloliquefaciens CH86-1

  • Kwon, Gun-Hee;Park, Jae-Yong;Kim, Jong-Sang;Lim, Jin-Kyu;Park, Cheon-Seok;Kwon, Dae-Young;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.515-518
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    • 2011
  • A gene encoding bacillopeptidase F, bpr86-1, was cloned from B. amyloliquefaciens CH86-1 isolated from cheonggukjang. This gene could encode a preproenzyme of 1,431 amino acids. When bpr86-1 was introduced into B. subtilis WB600 via pHY300PLK, an E. coli-Bacillus shuttle vector, the transformant showed fibrinolytic activity. During growth on LB, the fibrinolytic activity of cells increased sharply when they entered the stationary phase. The highest activity (761.4 mU/mg protein) was observed at 96 h of cultivation.

Cloning of aprE86-1 Gene Encoding a 27-kDa Mature Fibrinolytic Enzyme from Bacillus amyloliquefaciens CH86-1

  • Lee, Ae-Ran;Kim, Gyoung-Min;Kwon, Gun-Hee;Lee, Kang-Wook;Park, Jae-Yong;Chun, Ji-Yeon;Cha, Jae-Ho;Song, Young-Sun;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.370-374
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    • 2010
  • A gene encoding the major secreted fibrinolytic protein of Bacillus amyloliquefaciens CH86-1 was cloned from genomic DNAs. DNA sequencing showed that the gene, aprE86-1, could direct the synthesis of a mature protein 275 amino acids in length after processing. When aprE86-1 was introduced into B. subtilis, a mature 27-kDa protein was produced as expected. The fibrinolytic activity of the B. subtilis transformant (TF) was higher than that of B. amyloliquefaciens CH86-1, showing the possibility of increasing the fibrinolytic activity of Bacillus strains through genetic engineering.

Inhibition of Hepatocellular Carcinoma Cell Growth by the Extract of Symphytum offcinale L. and the Possible Mechanisms for this Inhibition

  • Ham, Seung-Shi;Park, Kyong-Gun;Lee, Yong-Moon;Lee, Young-Ik;Yoon, Ji-Won;Kim, Seong-Jin;Lee, euk-Sik
    • Preventive Nutrition and Food Science
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    • v.2 no.3
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    • pp.236-240
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    • 1997
  • A crude extract of Smphytum officinale L. (comfrey) was for its ability to inhibit he growth of hepatocellular carcinoma cells and expression of the insulin-like growth factor I (IGF-II) gene. The DNA synthesis of hepatocellular carcinoma cell lines, Hep G2, Hep 3B, and PLC/PRF/5 was inhibited by a crude extract of Smphytum officinale in both a time- and a dose-dependent manners. This plant extract also inhibited expression of the IGF-II gene. Since IGF-II exerts a mitogenic effect on Hep G2 cells, these results suggest that the growth inhibition by Symphytum officinale extract is, in part, mediated through the inhibition of IGF-II gene expression.

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