• 제목/요약/키워드: gene screening

검색결과 790건 처리시간 0.029초

Rheinheimera aquatica sp. nov., Antimicrobial Activity-Producing Bacterium Isolated from Freshwater Culture Pond

  • Chen, Wen-Ming;Lin, Chang-Yi;Young, Chiu-Chung;Sheu, Shih-Yi
    • Journal of Microbiology and Biotechnology
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    • 제20권10호
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    • pp.1386-1392
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    • 2010
  • A bacterial strain designated GR5$^T$, previously isolated from a freshwater culture pond in Taiwan while screening for bacteria for antimicrobial compounds, was characterized using a polyphasic taxonomic approach. Strain GR5$^T$ was found to be Gram-negative, aerobic, greenish-yellow colored, rod-shaped, and motile by means of a single polar flagellum. Growth occurred at $10-40^{\circ}C$ (optimum, $35^{\circ}C$), pH 7.0-8.0 (optimum pH 8.0), and with 0-2.0% NaCl (optimum, 0.5-1.0%). The major fatty acids were $C_{16:1}{\omega}7c$(36.3%), $C_{16:0}$(16.6%), $C_{12:0}$ 3-OH (12.5%), and $C_{18:1}{\omega}7c$(9.1%). The major respiratory quinone was Q-8, and the DNA G+C content of the genomic DNA was 51.9 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain GR5$^T$ belongs to the genus Rheinheimera, where its most closely related neighbors are Rheinheimera texasensis A62-14B$^T$ and Rheinheimera tangshanensis JA3-B52$^T$ with sequence similarities of 98.1% and 97.5%, respectively, and the sequence similarities to any other recognized species within Gammaproteobacteria are less than 96.5%. The mean level of DNA-DNA relatedness between strain GR5$^T$ and R. texasensis A62-14B$^T$, the strain most closely related to the isolate, was $26.5{\pm}7.6%$. Therefore, based on the phylogenetic and phenotypic data, strain GR5$^T$ should be classified as a novel species, for which the name Rheinheimera aquatica sp. nov. is proposed. The type strain is GR5$^T$ (=BCRC 80081$^T$=LMG 25379$^T$).

Systemic and Cell-Type Specific Profiling of Molecular Changes in Parkinson's Disease

  • Lee, Yunjong
    • Interdisciplinary Bio Central
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    • 제4권3호
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    • pp.6.1-6.12
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    • 2012
  • Parkinson's disease (PD) is a complicated neurodegenerative disorder although it is oftentimes defined by clinical motor symptoms originated from age dependent and progressive loss of dopaminergic neurons in the midbrain. The pathogenesis of PD involves dopaminergic and nondopaminergic neurons in many brain regions and the molecular mechanisms underlying the death of different cell types still remain to be elucidated. There are indications that PD causing disease processes occur in a global scale ranging from DNA to RNA, and proteins. Several PD-associated genes have been reported to play diverse roles in controlling cellular functions in different levels, such as chromatin structure, transcription, processing of mRNA, translational modulation, and posttranslational modification of proteins. The advent of quantitative high throughput screening (HTS) tools makes it possible to monitor systemic changes in DNA, RNA and proteins in PD models. Combined with dopamine neuron isolation or derivation of dopamine neurons from PD patient specific induced pluripotent stem cells (PD iPSCs), HTS techonologies will provide opportunities to draw PD causing sequences of molecular events in pathologically relevant PD samples. Here I discuss previous studies that identified molecular functions in which PD genes are involved, especially those signaling pathways that can be efficiently studied using HTS methodologies. Brief descriptions of quantitative and systemic tools looking at DNA, RNA and proteins will be followed. Finally, I will emphasize the use and potential benefits of PD iPSCs-derived dopaminergic neurons to screen signaling pathways that are initiated by PD linked gene mutations and thus causative for dopaminergic neurodegneration in PD.

Screening of Domestic Silkworm Strains for Efficient Heterologous Protein Expression by Bombyx mori Nuclear Polyhedrosis Virus (BmNPV)

  • Jo, Sun Jung;Choi, Ji-Hyun;Kang, Ju-Il;Lim, Jae-Hwan;Seok, Young Sik;Lee, Jae Man;Kusakabe, Takahiro;Hong, Sun Mee
    • International Journal of Industrial Entomology and Biomaterials
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    • 제29권2호
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    • pp.185-192
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    • 2014
  • Recombinant proteins can be generated quickly and easily in large amounts and at low-cost in silkworm larvae by using Bombyx mori nuclear polyhedrosis virus (BmNPV). We searched for high-permissive silkworm strains that have high production levels of heterologous proteins and are thus suitable for use as biofactories. In this study, we performed the analysis using a BmNPV vector expressing luciferase as a marker, and we confirmed protein expression by evaluating luciferase activity, determined by western blotting and luciferase ELISA, and confirmed transcription expression by semi- and quantitative real time PCR. For the selection of host silkworm strains, we first chose 52 domestic BmNPV sensitive strains and then identified 10 high-permissive and 5 low-permissive strains. In addition, to determine which hybrid of the high-permissive strains would show heterosis, nine strains derived through three-way crossing were tested for luciferase activity by western blotting, and luciferase ELISA. We found a correlation between luciferase activity and luciferase protein expression, but not transcription. There was no noticeable difference in protein expression levels between Jam313 as the high-permissive control strain and the three-way hybrid strains; however, the three-way cross strains showed lower luciferase activity compared with Jam313. In this study, luciferase protein production in the larvae of 52 domestic silkworm strains was elucidated using BmNPV.

의료정보 유출의 문제점과 의료정보보호 (The Medical Information Protection and major Issues)

  • 전영주
    • 한국컴퓨터정보학회논문지
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    • 제17권12호
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    • pp.251-258
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    • 2012
  • 과거 종이로 기록되던 환자의 의료정보는 점점 현대과학의 눈부신 진화와 발전으로 현재는 종이를 대신하는 전자적 시스템인 전자의무기록시스템 형태로 발전되었다. 또한 병원에서 활용되는 모든 의료정보들이 전자적시스템을 이용하여 활용되고 있는 것이 현실이다. 그러나 과학기술의 발전과 더불어 의료정보의 피해 사례는 계속 증가할 것으로 보인다. 보험사기 예방을 위해 의료정보를 공유해야한다는 보험단체들의 주장, 개인정보를 얻기 위해 금전적 불법거래가 성행되고 있다는 뉴스 등을 접할때면 의료정보 보호에 관한 대책 마련이 절실함을 의미한다. 따라서 의료정보와 관련된 유출상의 문제점을 정확히 파악하는 것은 의료정보 유출의 부작용을 예방하고 의료정보 보호와 미비점을 보완하는 선결과제인 것이다. 따라서 현재의 의료법 및 개인정보보호법제만으로는 의료정보의 표준화와 의료정보보호 등을 규율할 수 없고, 의료정보에서 발생되는 여러 문제들을 보호하기에는 미흡하다고 판단된다. 따라서 본 논문은 미흡한 의료정보관련 법제의 연구를 위한 선결 과제로 의료정보의 유출로 발생될 수 있는 문제점을 파악하는 것이 무엇보다 중요하다고 판단되며, 이에 의료정보의 유출로 인한 문제점을 파악하고 의료정보를 보호할 수 있는 입법방안을 중심으로 연구하고자 한다.

In Silico Screening for Angiogenesis-Related Genes in Rat Astrocytes

  • Kim, Soo-Young;Lee, Sae-Won;You, Sung Yong;Rha, Sun Young;Kim, Kyu-Won
    • Genomics & Informatics
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    • 제2권1호
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    • pp.36-44
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    • 2004
  • Astrocytes play supportive roles for neurons in the brain. Recently, they have been accepted to have various functions in the vascular system as well as in the nervous system. We investigated the differential gene expression in rat astrocytes according to the oxygen tension, which is a crucial factor for angiogenesis. A cDNA microarray was performed to find the genes whose expression was sensitive to oxygen tension. We found 26 genes in the astrocyte were found and classified into 4 groups. In order to show the genes' relevancy to angiogenesis, seven of the 26 genes were investigated to see whether they have capabilities of interaction with angiogenesis­related factors in AngioDB. Through this investigation, we found interactions of three proteins with angiogenesis-related factors. These genes were further investigated with a new focus on the vascular endothelial growth factor (VEGF) expression in an astrocyte based on our hypothesis that astrocytes can have effects on endothelial angiogenesis via the release of VEGF. Collectively, we identified several genes whose expressions were dependent on the oxygen concentration of the astrocyte. Furthermore, the relevancy of astrocytes to angiogenesis was investigated using preexisting information of AngioDB, and suggested a possible signaling pathway for VEGF expression in the aspects of brain endothelial angiogenesis by astrocytes.

방사선을 이용한 벼 흰잎마름병 저항성 돌연변이 벼 계통의 선발 (Screening of Gamma Radiation-Induced Pathogen Resistance Rice Lines against Xanthomonas oryzae pv. oryzae)

  • 임찬주;이하연;김웅범;아마드 라자;문제선;김동섭;권석윤
    • 방사선산업학회지
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    • 제4권3호
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    • pp.209-213
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    • 2010
  • Bacterial blight is one of the most serious diseases of rice (Oryza sativa L.), and it has been known that Xanthomonas oryzae pv. oryzae (Xoo) causes this disease symptom. To develop resistance rice cultivars against Xoo, 3,000 lines of $M_3$ mutants, which were irradiated with gamma ray, were tested by 'scissor-dip method' primarily, and 191 putative resistant lines were selected. In $M_4$ generation, these lines were screened again with various ways such as measuring of symptom of bacterial blight in leaf, number of tiller, fresh weight, and phenotypic segregation ratio in next generation. Finally, six resistance lines were selected. RT-PCR analysis revealed that these lines displayed high level of R-genes such as Xa21, Pi36, and Pi-ta. These results indicate that mutations by gamma ray cause disruptions of regulatory signal transduction systems of these R-genes. Furthermore, these selected mutants could be useful for the development of rice cultivar resistant to Xoo.

Ginsenoside Rh2 upregulates long noncoding RNA STXBP5-AS1 to sponge microRNA-4425 in suppressing breast cancer cell proliferation

  • Park, Jae Eun;Kim, Hyeon Woo;Yun, Sung Hwan;Kim, Sun Jung
    • Journal of Ginseng Research
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    • 제45권6호
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    • pp.754-762
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    • 2021
  • Background: Ginsenoside Rh2, a major saponin derivative in ginseng extract, is recognized for its anti-cancer activities. Compared to coding genes, studies on long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) that are regulated by Rh2 in cancer cells, especially on competitive endogenous RNA (ceRNA) are sparse. Methods: LncRNAs whose promoter DNA methylation level was significantly altered by Rh2 were screened from methylation array data. The effect of STXBP5-AS1, miR-4425, and RNF217 on the proliferation and apoptosis of MCF-7 breast cancer cells was monitored in the presence of Rh2 after deregulating the corresponding gene. The ceRNA relationship between STXBP5-AS1 and miR-4425 was examined by measuring the luciferase activity of a recombinant luciferase/STXBP5-AS1 plasmid construct in the presence of mimic miR-4425. Results: Inhibition of STXBP5-AS1 decreased apoptosis but stimulated growth of the MCF-7 cells, suggesting tumor-suppressive activity of the lncRNA. MiR-4425 was identified to have a binding site on STXBP5-AS1 and proven to be downregulated by STXBP5-AS1 as well as by Rh2. In contrast to STXBP5-AS1, miR-4425 showed pro-proliferation activity by inducing a decrease in apoptosis but increased growth of the MCF-7 cells. MiR-4425 decreased luciferase activity from the luciferase/STXBP5-AS1 construct by 26%. Screening the target genes of miR-4425 and Rh2 revealed that Rh2, STXBP5-AS1, and miR-4425 consistently regulated tumor suppressor RNF217 at both the RNA and protein level. Conclusion: LncRNA STXBP5-AS1 is upregulated by Rh2 via promoter hypomethylation and acts as a ceRNA, sponging the oncogenic miR-4425. Therefore, Rh2 controls the STXBP5-AS1/miR-4425/RNF217 axis to suppress breast cancer cell growth.

Antifungal Mechanism of Action of Lauryl Betaine Against Skin-Associated Fungus Malassezia restricta

  • Do, Eunsoo;Lee, Hyun Gee;Park, Minji;Cho, Yong-Joon;Kim, Dong Hyeun;Park, Se-Ho;Eun, Daekyung;Park, Taehun;An, Susun;Jung, Won Hee
    • Mycobiology
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    • 제47권2호
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    • pp.242-249
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    • 2019
  • Betaine derivatives are considered major ingredients of shampoos and are commonly used as antistatic and viscosity-increasing agents. Several studies have also suggested that betaine derivatives can be used as antimicrobial agents. However, the antifungal activity and mechanism of action of betaine derivatives have not yet been fully understood. In this study, we investigated the antifungal activity of six betaine derivatives against Malassezia restricta, which is the most frequently isolated fungus from the human skin and is implicated in the development of dandruff. We found that, among the six betaine derivatives, lauryl betaine showed the most potent antifungal activity. The mechanism of action of lauryl betaine was studied mainly using another phylogenetically close model fungal organism, Cryptococcus neoformans, because of a lack of available genetic manipulation and functional genomics tools for M. restricta. Our genome-wide reverse genetic screening method using the C. neoformans gene deletion mutant library showed that the mutants with mutations in genes for cell membrane synthesis and integrity, particularly ergosterol synthesis, are highly sensitive to lauryl betaine. Furthermore, transcriptome changes in both C. neoformans and M. restricta cells grown in the presence of lauryl betaine were analyzed and the results indicated that the compound mainly affected cell membrane synthesis, particularly ergosterol synthesis. Overall, our data demonstrated that lauryl betaine influences ergosterol synthesis in C. neoformans and that the compound exerts a similar mechanism of action on M. restricta.

Antimicrobial Efficacy of Leuconostoc spp. Isolated from Indian Meat against Escherichia coli and Listeria monocytogenesin Spinach Leaves

  • Thangavel, Gokila;Subramaniyam, Thiruvengadam
    • 한국축산식품학회지
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    • 제39권4호
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    • pp.677-685
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    • 2019
  • Five Leuconostoc strains (CM17, CM19, PM30, PM32, and PM36) previously isolated from Indian meat showed promising antimicrobial activity against food pathogens in screening assay. This study evaluates the efficacy of these isolates against Escherichia coli Microbial Type Culture Collection and Gene Bank (MTCC) 443 and Listeria monocytogenes (MTCC 657) in spinach leaves. Challenge studies were conducted by inoculating E. coli and L. monocytogenes at 6 to 7 $Log_{10}CFU/g$ of the leaves respectively and treating them with cell free supernatant (CFS) of 48 h cultures of the isolates. The samples were stored at $4^{\circ}C$ and analyzed over a period of 5 d. The study was conducted in triplicates and statistical analysis was carried out using one-way Anova. The counts of the pathogens did not increase over the 5 d period in the control samples, without any treatment. Whereas in the case of CFS treatments, significant reduction (p<0.05) was observed in both E. coli and L. monocytogenes from 1 to 5 d with all the 5 strains as compared to the control. The counts of Listeria dropped by 0.5 to 1 log by 5 d, with PM 36 showing the highest reduction (1 log). In the case of E. coli, 1.1 to 1.5 log reduction was observed by 5 d, with again PM 36 showing the highest reduction (1.5). The overall results indicate that the isolates (specifically PM36) not only showed efficacy in in vitro studies but are also proved to be effective in food matrix making them potential clean label antimicrobial alternatives for food application.

Novel Discovery of LINE-1 in a Korean Individual by a Target Enrichment Method

  • Shin, Wonseok;Mun, Seyoung;Kim, Junse;Lee, Wooseok;Park, Dong-Guk;Choi, Seungkyu;Lee, Tae Yoon;Cha, Seunghee;Han, Kyudong
    • Molecules and Cells
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    • 제42권1호
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    • pp.87-95
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    • 2019
  • Long interspersed element-1 (LINE-1 or L1) is an autonomous retrotransposon, which is capable of inserting into a new region of genome. Previous studies have reported that these elements lead to genomic variations and altered functions by affecting gene expression and genetic networks. Mounting evidence strongly indicates that genetic diseases or various cancers can occur as a result of retrotransposition events that involve L1s. Therefore, the development of methodologies to study the structural variations and interpersonal insertion polymorphisms by L1 element-associated changes in an individual genome is invaluable. In this study, we applied a systematic approach to identify human-specific L1s (i.e., L1Hs) through the bioinformatics analysis of high-throughput next-generation sequencing data. We identified 525 candidates that could be inferred to carry non-reference L1Hs in a Korean individual genome (KPGP9). Among them, we randomly selected 40 candidates and validated that approximately 92.5% of non-reference L1Hs were inserted into a KPGP9 genome. In addition, unlike conventional methods, our relatively simple and expedited approach was highly reproducible in confirming the L1 insertions. Taken together, our findings strongly support that the identification of non-reference L1Hs by our novel target enrichment method demonstrates its future application to genomic variation studies on the risk of cancer and genetic disorders.