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Novel Vancomycin Resistance System in Streptomyces coelicolor

  • Hong, Hee-Jeon
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2005.05a
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    • pp.143-147
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    • 2005
  • The non-pathogenic, non-glycopeptide-producing actinomycete Streptomyces coelicolor carries a cluster of seven genes (vanSRJKHAX) that confers inducible, high-level resistance to vancomycin. The van genes are organised into four transcription units, vanRS, vanJ, vanK and vanHAX, and these transcripts are induced by vancomycin in a vanR-dependent manner. vanHAX are orthologuous to genes found in vancomycin resistant enterococci that encode enzymes predicted to reprogramme peptidoglycan biosynthesis such that cell wall precursors terminate in D-Ala-D-Lac, rather than D-Ala-D-Ala. vanR and vanS encode a two-component signal transduction system that mediates transcriptional induction of the seven van genes. vanJ and vanK are novel genes that have no counterpart in previously characterised vancomycin-resistance clusters from pathogens. VanK is essential for vancomycin resistance in S. coelicolor and it is required for adding Gly branch to stem peptides terminating D-Ala-D-Lac. Because VanK can recognise D-Lac-containing precursors but the constitutively expressed femX enzyme, encoded elsewhere on the chromosome, cannot recognize D-Lac-containing precursors as a substrate, vancomycin-induced expression of VanHAX in a vanK mutant is lethal. Further, femX null mutants are viable in the presence of glycopeptide antibiotics but die in their absence. Bioassay using vanJp-neo fusion reporter system also showed that all identified inducers for van genes expression were glycopeptide antibiotics, but teicoplanin, a membrane-anchored glycopeptide, failed to act as an inducer.

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A Study on the Professional Image Concept for Appearance Management in the Workplace (Part 1) (직장인의 외모관리를 위한 프로페셔널 이미지 개념 연구 (제1보))

  • Yoo, Hee;Choi, Sun-Hyung
    • Journal of the Korean Society of Clothing and Textiles
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    • v.34 no.10
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    • pp.1742-1753
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    • 2010
  • This study clarifies the importance, advantages, and constituents of the professional image by appearance management in the workplace. In-depth interviews were conducted with 8 participants who have over 8 years experience in their respective fields. The transcripts of the interview were classified according to a card classification method. The results of the study were as follows: First, the professional image by appearance management is important, because there is a special treatment by image in interaction on business. Second, the advantages of the professional image were to achieve a career opportunity, improve interpersonal relationships, obtain special treatment or protection from a biased angle, and increase business reliability. Third, the constituents of the professional image were conscientiousness and self-management skills, assertive and open attitudes, leadership, interpersonal skills, customer-satisfaction spirit, expert knowledge and skills, and confidence in the business results.

Generation of Expressed Sequence Tags for Immune Gene Discovery and Marker Development in the Sea Squirt, Halocynthia roretzi

  • Kim, Young-Ok;Cho, Hyun-Kook;Park, Eun-Mi;Nam, Bo-Hye;Hur, Young-Baek;Lee, Sang-Jun;Cheong, Jae-Hun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1510-1517
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    • 2008
  • Expresssed sequence tag (EST) analysis was developed from three cDNA libraries constructed from cells of the digestive tract, gonad, and liver of sea squirt. Randomly selected cDNA clones were partially sequenced to generate a total of 922 ESTs, in which 687 unique ESTs were identified respectively. Results of BLASTX search showed that 612 ESTs (89%) have homology to genes of known function whereas 75 ESTs (11%) were unidentified or novel. Based on the major function of their encoded proteins, the identified clones were classified into ten broad categories. We also identified several kinds of immune-related genes as identifying novel genes. Sequence analysis of ESTs revealed the presence of microsatellite-containing genes that may be valuable for further gene mapping studies. The accumulation of a large number of identified cDNA clones is invaluable for the study of sea squirt genetics and developmental biology. Further studies using cDNA microarrays are needed to identify the differentially expressed transcripts after disease infection.

Expression of temperature responsive genes in cell cultures derived from Bombyx mori

  • Kim, Eun-Young;Kang, Min-Uk;Park, Kwan-Ho;Choi, Kwang-Ho;Nho, Si-Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.2
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    • pp.95-102
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    • 2015
  • Insects are heterotherms that exhibit a close relationship between their ecology (especially temperature changes) and physiology. In the present study, selected genes associated with cell death and temperature were examined to determine gene expression in Bombyx mori in high and low temperature environments. We determined the amount of dsRNA, different concentrations of dsRNA, and different type of cells to set the conditions most efficient for RNAi. We then prepared dsRNA transcripts of the genes associated with cell death and temperature response. We analyzed cell damage via Trypan blue staining and found that cell viability was reduced after knockdown of these genes. The special transduced cell lines produced in the present study can be applied in various research fields. We also expect that these cell lines can be used as a research tool for the precise functional analysis of various genes.

Gene Expression Profiling in Rice Infected with Rice Blast Fungus using SAGE

  • Kim, Sang-Gon;Kim, Sun-Tae;Kim, Sung-Kun;Kang, Kyu-Young
    • The Plant Pathology Journal
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    • v.24 no.4
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    • pp.384-391
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    • 2008
  • Rice blast disease, caused by the pathogenic fungus Magnaporthe grisea, is a serious issue in rice (Oryza sativa L.) growing regions of the world. Transcript profiling in rice inoculated with the fungus has been investigated using the transcriptomics technology, serial analysis of gene expression (SAGE). Short sequence tags containing sufficient information which are ten base-pairs representing the unique transcripts were identified by SAGE technology. We identified a total of 910 tag sequences via the GenBank database, and the resulting genes were shown to be up-regulated in all functional categories under the fungal biotic stress. Compared to the compatible interaction, the stress and defense genes in the incompatible interaction appear to be more up-regulated. Particularly, thaumatin-like gene (TLP) was investigated in determining the gene and protein expression level utilizing Northern and Western blotting analyses, resulting in an increase in both the gene and the protein expression level which arose earlier in the incompatible interaction than in the compatible interaction.

Application of Epstein-Barr Virus Cell Lines (CCL85 EB-3) in Performing the EBER mRNA In Situ Hybridization as a Positive Control (Epstein-Barr 바이러스 인사이투 보결합 시행시 양성대조표지로서의 버키트 림프종 세포주 (CCL85 EB-3)의 응용)

  • Kim, Sung-Sook;Han, Woon-Sup;Suh, Joo-Young;Huh, Joo-Ryung
    • The Korean Journal of Cytopathology
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    • v.7 no.1
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    • pp.38-43
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    • 1996
  • Epstein-Barr virus(EBV) is associated with a wide spectrum of benign and malignant disorders including leukoplakia, Hodgkln's lymphoma, central nervous system lymphoma, peripheral T cell lymphoma and nasopharyngeal undifferentiated carcinoma. There are several distinctive aspects of biology of the virus that are important in investigation of virus in clinical specimens. The abundant expression of the EBER mRNA transcripts makes possible the sensitive detection of latent expression in EBV-associated tumors. Although there has been a dramatic increased interest in the direct characterization of EBV in clinical specimens, there have been few studios about the effective and reliable positive controls in performing in situ hybridization technique for EBV, especially on paraffin-em bedded tissue. We applied Burkitts lymphoma ceil line as positive control in EBV in situ hydridization using Oncor Kit. The cell block of Burkitt lymphoma cell line(CCL85 EB-3) showed strong and specific positivity for EBER in situ in nuclei of EBV infected cells.

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Re-Engineering of Carcinoembryonic Antigen RNA with the Group I Intron of Tetrahymena thermophila by Targeted Trans-Splicing

  • JUNG HEUNG-SU;LEE SEONG-WOOK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1408-1413
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    • 2005
  • Elevated expression of carcinoembryonic antigen (CEA) has been implicated in various biological aspects of neoplasia such as tumor cell adhesion, metastasis, blocking of cellular immune mechanisms, and antiapoptosis function. Thus, the CEA could be an important target for anticancer therapy. In this study, we developed Tetrahymena group 1 intron-based trans-splicing ribozymes that can specifically target and replace CEA RNA. To this end, we first determined which regions of the CEA RNA were accessible to ribozymes by employing an RNA mapping strategy that was based on a trans-splicing ribozyme library. Next, we assessed the ribozyme activities by comparing the trans-splicing activities of several ribozymes that targeted different regions of the CEA RNA, and then the ribozyme that could target the most accessible site was observed to be the most active with high fidelity in vitro. Moreover, the specific trans-splicing ribozyme was found to react with and altered the target CEA transcripts in mammalian cells with high fidelity. These results suggest that the Tetrahymena ribozyme can be utilized to replace CEA RNAs in tumors with a new RNA-harboring anticancer activity, thereby hopefully reverting the malignant phenotype.

Identification of the Vibrio vulnificus cadC and Evaluation of Its Role in Acid Tolerance

  • Rhee, Jee-Eun;Ju, Hyun-Mok;Park, U-Ryung;Park, Byoung-Chul;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1093-1098
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    • 2004
  • An open reading frame encoding CadC, consisting of 526 amino acids, was identified from the upstream region of the Vibrio vulnificus cadBA operon. The deduced amino acid sequences of the cadC were 22 to 78% similar to those reported from other Enterobacteriaceae. Functions of cadC gene on acid tolerance were assessed by comparing acid tolerances of V. vulnificus and its isogenic mutant, whose cadC gene was inactivated by allelic exchanges. The results demonstrated that the gene product of cadC contributes to acid tolerance of V. vulnificus, and that its contribution is dependent on prior exposure of cells to moderately acidic pH. The cellular level of cadB and cadA transcripts decreased in the cadC mutant, indicating that CadC exerts its effect on acid tolerance of V. vulnificus by enhancing the expression of cadBA in a pH-dependent manner.

Cloning and Characterization of DAP10 homologue gene from Olive Flounder, Paralichthys olivaceus

  • Park, Chan-Il;Kim, Mu-Chan;Hwang, Jee-Youn;Kim, Ki-Hyuk;Kim, Joo-Won
    • Journal of fish pathology
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    • v.19 no.3
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    • pp.227-233
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    • 2006
  • Olive flounder immunoreceptor DAP10 homologue cDNA was cloned from a peripheral blood lymphocytes (PBLs) cDNA library. The length of the olive flounder DAP10 cDNA is 473bp and it contains an open reading frame of 234bp. The predicted polypeptide sequence is 78 amino acids, consisting of a 22-amino acid leader, an 11-amino acid extracellular domain, a 21-amino acid transmembrane segment, and a 24-amino acid cytoplasmic domain. The amino acid sequence of olive flounder DAP10 has 56%, 50%, 32%, 31%, and 31% sequence identity with zebrafish DAP10, catfish DAP10, cattle DAP10, rat DAP10 and Monkey DAP10, respectively. Olive flounder DAP10 has a conserved aspartic acid in the transmembrane domain and a phophatidylinositol-3 kinase-binding site (YxxM/V) in the cytoplasmic region. Genomic organization reveals that olive flounder DAP10 comprises five exons and four introns. A phylogenetic analysis based on the deduced amino acid sequence grouped the olive flounder DAP10 with other species DAP10. In RT-PCR analysis, DAP10 transcripts were detected predominantly in PBLs, kidney, spleen and intestine.

Physiological and Molecular Responses of Maize to High Temperature Stress During Summer in the Southern Region of Korea

  • Lee, Joon-Woo;Min, Chang-Woo;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.38 no.3
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    • pp.170-174
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    • 2018
  • Environmental stresses caused by climate change, such as high temperature, drought and salinity severely impact plant growth and productivity. Among these factors, high temperature stress will become more severe during summer. In this study, we examined physiological and molecular responses of maize plants to high temperature stress during summer. Highest level of $H_2O_2$ was observed in maize leaves collected July 26 compared with June 25 and July 12. Results indicated that high temperature stress triggers production of reactive oxygen species (ROS) in maize leaves. In addition, photosynthetic efficiency (Fv/Fm) sharply decreased in leaves with increasing air temperatures during the day in the field. RT-PCR analysis of maize plants exposed to high temperatures of during the day in field revealed increased accumulation of mitochondrial and chloroplastic small heat shock protein (HSP) transcripts. Results demonstrate that Fv/Fm values and organelle-localized small HSP gene could be used as physiological and molecular indicators of plants impacted by environmental stresses.