• Title/Summary/Keyword: sM gene

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Amino acid substitutions conferring cold-sensitive phenotype on the yeast MTF1 gene

  • Jang, Sei-Heon
    • Journal of Microbiology
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    • v.35 no.3
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    • pp.228-233
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    • 1997
  • The MTF1 gene of Saccharomyces cerevisiae encodes a 43 kDa MITOCHONDRIAL RNA polymerase specificity factor which recognizes mitochondrial promoters to initiate correct transcription. To better understand structure-function of the MTF1 gene as well as the transcription mechanism of mitochondrial RNA polymerase, two cold-sensitive alleles of the MTF1 mutation were isolated by plasmid shuffling method after PCR-based random mutagenesis of the MTF1 gene. The mutation sites were analyzed by nucleotide sequencing. These cs phenotype mtf1 mutants were respiration competent on the nonfermentible glycerol medium at the permissive temperature, but incompetent at 13.deg.C. The cs phenotype allele of the MTF1, yJH147, encoded an L146P replacement. The other cs allele, yJH148, contained K179E and K214M double replacements. Mutations in both alleles were in a region of Mtflp which is located between domains with amino acid sequence similarities to conserved regions 2 and 3 of bacterial s factors.

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Inhalation Delivery of Nano-Aerosol Containing PEI-glucose-PTEN Complex Induced Change of Protein Translation in Kras Knock-Qut Lung Cancer Model Mice

  • Kim, H. W.;Park, I. K.;C. S. Cho;M. H. Cho
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2003.10b
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    • pp.163-163
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    • 2003
  • Difficulties of long-tenn survival of lung cancer patients treated with conventional therapies require the need for novel approaches and gene therapy holds promise in this area. Several genes are known to have anti-tumor activities and have been used as a gene of delivery, however, a number of problems such as efficiency, specificity of the gene delivery hinder the application of gene therapy.(omitted)

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Emergence of CTX-M-15 Extended Spectrum β-lactamase and ArmA-Producing Enterobacter cloacae (CTX-M-15형 Extended Spectrum β-lactamase와 ArmA 동시 생성 Enterobacter cloacae의 출현)

  • Sung, Ji-Youn
    • Journal of Digital Convergence
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    • v.13 no.12
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    • pp.313-318
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    • 2015
  • We investigated the prevalence of extended spectrum ${\beta}$-lactamase (ESBL) genes and 16S rRNA methyltransferase genes to study antimicrobial resistance mechanisms of Enterobacter cloacae strains isolated from a university hospital in the Chungcheong province of Korea. Eight of the bacteria strains involved in this study contained CTX-M-15 type ESBL. Among 8 strains harboring the ESBL gene, 3 strains also harbored armA gene. The three isolates showed resistance to antimicrobial agents belonged to third cephalosporin, aminoglycoside, and fluoroquinolones. Furthermore, interspecies plasmid transfer of the antimicrobial resistant genes may induced horizontal spreading of the genes and emergence of multidrug resistant bacteria. Therefore, surveillance for existence of antimicrobial resistance determinants is important to prevent distribution of antimicrobial resistant strains.

Isolation and Characterization of Zymomonas mobilis DNA Fragments Showing Promoter Activity in Escherichia coli (Escherichia coli에서 Promoter 활성을 보이는 Zymomonas mobilis DNA 조각의 분리와 분석)

  • Kim, Eun-Joon;Yoon, Ki-Hong;M.Y. Pack
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.600-605
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    • 1989
  • For the purpose of isolation of the Zymomonas mobilis DNA fragments showing promoter activity in Escherichia coli, a promoter screening vector, PCMT215 was constructed by transferring a promoterless chloramphenicol acetyltransferase (CAT) gene of pYEJ001 into pMT21 which contains $\beta$-lactamase gene and multiple cloning sites. A library of Z, mobilis Sau3AI DNA fragments was constructed in E. coli using the newly constructed pCMT215. Fourteen clones showing resistance to chloramphenicol ranging in concentration from 30 to 750 $\mu$g/$m\ell$ were selected. From five clones of them, the Z. mobilis DNA fragments expressing CAT gene of the recombinant plasmids were sequenced and then sites of transcriptional initiation were identified. The nucleotide sequences of the cloned DNA shared AT rich regions, poly A's or T's stretches and palindromic regions. The positions of transcriptional initiation for CAT gene occurred at more than one site spaced over by 4 to 190 base pairs on the cloned fragments in E. coli.

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Classification of Gene Data Using Membership Function and Neural Network (소속 함수와 유전자 정보의 신경망을 이용한 유전자 타입의 분류)

  • Yeom, Hae-Young;Kim, Jae-Hyup;Moon, Young-Shik
    • Journal of the Institute of Electronics Engineers of Korea CI
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    • v.42 no.4 s.304
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    • pp.33-42
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    • 2005
  • This paper proposes a classification method for gene expression data, using membership function and neural network. The gene expression is a process to produce mRNA and protains which generate a living body, and the gene expression data is important to find out the functions and correlations of genes. Such gene expression data can be obtained from DNA 칩 massively and quickly. However, thousands of gene expression data may not be useful until it is well organized. Therefore a classification method is necessary to find the characteristics of gene data acquired from the gene expression. In the proposed method, a set of gene data is extracted according to the fisher's criterion, because we assume that selected gene data is the well-classified data sample. However, the selected gene data does not guarantee well-classified data sample and we calculate feature values using membership function to reduce the influence of outliers in gene data. Feature vectors estimated from the selected feature values are used to train back propagation neural network. The experimental results show that the clustering performance of the proposed method has been improved compared to other existing methods in various gene expression data.

Endo-sulfatase Sulf-1 Protein Expression is Down-regulated in Gastric Cancer

  • Gopal, Gopisetty;Shirley, Sundersingh;Raja, Uthandaraman Mahalinga;Rajkumar, Thangarajan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.641-646
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    • 2012
  • In our recent report on gene expression in gastric cancer we identified the endo-sulfatase Sulf-1 gene to be up-regulated in gastric tumors relative to apparently normal (AN), and paired normal (PN) gastric tissue samples. In the present report we investigate the protein expression levels of Sulf-1 gene in gastric tumors, AN and PN samples using tissue microarray (TMA) and immunohistochemistry. Expression data was collected from two sets of TMA's containing replicate sections of tissue samples. Scoring data from TMA set-1 revealed a significant difference in Sulf-1 immunoreactivity between tumors and "normals" (PN and AN) (p-value = 0.001928). Also, Sulf-1 expression in tumors was also significantly different from either PN (p-value = 0.019) or AN (p-value = 0.006) samples. Similar results were obtained from analysis of scoring data from the second set of arrays. Comparison of mRNA expression and protein expression in gastric tumor tissues revealed that in 6/20 (30%) tumor samples showed up-regulated protein expression concordant with over-expression of mRNA. However, a discord with mRNA being over-expressed relative to down regulated protein expression was observed in majority 14/20 (70%) of tumor samples. Our study indicates down regulation of Sulf-1 protein expression in gastric tumors relative to PN and AN samples which is discordant with mRNA over-expression seen in tumors.

Cosuppression and RNAi induced by Arabidopsis ortholog gene sequences in tobacco

  • Oka, Shin-Ichiro;Midorikawa, Kaoru;Kodama, Hiroaki
    • Plant Biotechnology Reports
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    • v.4 no.3
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    • pp.185-192
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    • 2010
  • The Arabidopsis ${\omega}$-3 fatty acid desaturase (AtFAD7) catalyzes the synthesis of trienoic fatty acids (TA). A transgenic tobacco line, T15, was produced by a sense AtFAD7 construct and showed a cosuppression-like phenotype, namely extremely low TA levels. The sequence similarity between AtFAD7 and a tobacco ortholog gene, NtFAD7, was moderate (about 69%) in the coding sequences. AtFAD7 siRNAs accumulated at a high level, and both AtFAD7 and NtFAD7 mRNAs are degraded in T15 plants. The low-TA phenotype in T15 was dependent on a tobacco RNA-dependent RNA polymerase6 (NtRDR6). We also produced tobacco RNAi plants targeting AtFAD7 gene sequences. The AtFAD7 siRNA level was trace, which was associated with a slight reduction in leaf TA level. Unexpectedly, this RNAi plant showed an increased NtFAD7 transcript level. To investigate the effect of translational inhibition on stability of the NtFAD7 mRNAs, leaves of the wild-type tobacco plants were treated with a translational inhibitor, cycloheximide. The level of NtFAD7 mRNAs significantly increased after cycloheximde treatment. These results suggest that the translational inhibition by low levels of AtFAD7 siRNAs or by cycloheximide increased stability of NtFAD7 mRNA. The degree of silencing by an RNAi construct targeting the AtFAD7 gene was increased by co-existence of the AtFAD7 transgene, where NtRDR6-dependent amplification of siRNAs occurred. These results indicate that NtRDR6 can emphasize silencing effects in both cosuppression and RNAi.

MICROPATTERNED GROOVES AND ACID-ETCHING ON TITANIUM SUBSTRATA ALTER VIABILITY AND GENE EXPRESSION OF ADHERED HUMAN GINGIVAL FIBROBLASTS: A PILOT STUDY

  • Lee, Suk-Won;Kim, Su-Yeon;Lee, Keun-Woo
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.3
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    • pp.375-381
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    • 2007
  • Statement of problem. Prior to determining an optimal width of micropatterned grooves provided on titanium substrata, we have done a pilot study using surface topographies in combined microm and submicrom levels. Purpose. The purpose of this study was twofold 1) to assess the proliferation and 2) to analyze the expression of genes encoding the intracellular signaling proteins involved in cell-substratum adhesions and adhesion-dependent G1 phase cell cycle progression of human gingival fibroblasts plated on smooth and microgrooved/acid-etched titanium substrata. Material and methods. Three groups of titanium discs as NE0 (smooth Ti substrata), E15 (Ti substrata with microgrooves of $15{\mu}m$ of spacing and $3.5{\mu}m$ in depth and with further acidetching), and E30 (Ti substrata with microgrooves of $30{\mu}m$ spacing and $3.5{\mu}m$ in depth and with further acid-etching) served as the human gingival fibroblasts' substrata. Viability and proliferation of fibroblasts were determined using an XTT assay. Gene expressions of fibronectin, ${\alpha}5$ integrin, CDK4, and $p27^{kip}$ were analyzed in RT-PCR. Cell-substratum interactions were analyzed in SEM. Results. From the XTT assay at 24 h incubation, the mean optical density (OD) value of E15 was significantly greater than the values of E30 and NE0. At 48 and 96 h however, the mean OD values of E30 were significantly greater than the values of E15 and NE0. No differences in the expression of PCR transcripts at 96 h incubations were noted between groups, whereas at 48 h, an unexpected increase in the expression of all the transcripts were noted in E15 compared with other two groups. Fibroblasts were observed to orient and adhere inside the microgrooves. Conclusion. Micropatterned grooves and acid-etching on Ti substrata alter viability and gene expression of adhered human gingival fibroblasts.

Production of Bovine Transgenic Embryos Derived from Non-transfected and Transfected Adult Cells (외부유전자가 도입된 체세포를 이용한 소 형질전환 복제란 생산)

  • J. K. Cho;M.M.U. Bhuiyan;G. Jang;Park, E. S.;J. M. Lim;S. K. Kang;Lee, B. C.;W. S. Hwang
    • Journal of Embryo Transfer
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    • v.17 no.2
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    • pp.109-115
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    • 2002
  • The present study was conducted for the production of transgenic cloned cows those secrete human lactoferricin into milk by somatic cell nuclear transfer (NT). To estimate detrimental effects of gene transfection on transgenic cloned embryo production, development rates of NT embryos were compared between transfected and non-transfected cumulus and ear fibroblast cells. An expression plasmid for human lactofericin (pbeta-LFC) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and human lactoferricin target gene into a pcDNA3 plasmid. Two bovine somatic cell lines (cumulus cell and ear fibroblast) were established and transfected with the expression plasmid using a liposomal transfection reagent, Fugene6 as a carrier. Cumulus cell and ear fibroblast were transfected at the passage of 2 to 4, trypsinized and GFP-expressing cells were randomly selected and used for somatic cell NT. Developmental competences (rates of fusion, cleavage, and blastocyst formation) in bovine transgenic somatic cell NT embryos reconstructed with non-transfectecd cells were significantly higher than those from transfected cells in cumulus cell and ear fibroblast (P<0.05). This study indicated that transfection of done. cell has detrimental effect on embryo development in bovine transgenic NT.

Histone deacetylation effects of the CYP1A1 promoter activity, proliferation and apoptosis of cells in hepatic, prostate and breast cancer cells

  • K.N. Min;K.E. Joung;M.J. Cho;J.Y. An;Kim, D.K.;Y.Y. Sheen
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.91-91
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    • 2003
  • We have studied the mechanism of action of TCDD on CYP1A1 promoter activity in both Hepa I and MCF-7 cells using transient transfection system with plAl-Luc reporter gene. When HDAC inhibitors, such as trichostatin A, HC toxin and a novel HDAC inhibitor, IN2001 were cotreated with TCDD to the cells transfected with plAl-Luc reporter gene, the basal promoter activity of CYP1A1 was increased by HDAC inhibitors. Also, in MCF-7 human breast cancer cells, HDAC inhibitors, such as IN2001 and trichostatin A increased the basal activity of CYP1A1 promoter but TCDD stimulated CYP1A1 promoter activity was not changed by HDAC inhibitors. And, in stably-transfected Hepa I cells with plAl-Luc, HDAC inhibitors increased the basal promoter activity only.

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