• Title/Summary/Keyword: DnaB

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Inhibition of the DevSR Two-Component System by Overexpression of Mycobacterium tuberculosis PknB in Mycobacterium smegmatis

  • Bae, Hyun-Jung;Lee, Ha-Na;Baek, Mi-Na;Park, Eun-Jin;Eom, Chi-Yong;Ko, In-Jeong;Kang, Ho-Young;Oh, Jeong-Il
    • Molecules and Cells
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    • v.40 no.9
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    • pp.632-642
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    • 2017
  • The DevSR (DosSR) two-component system, which is a major regulatory system involved in oxygen sensing in mycobacteria, plays an important role in hypoxic induction of many genes in mycobacteria. We demonstrated that overexpression of the kinase domain of Mycobacterium tuberculosis (Mtb) PknB inhibited transcriptional activity of the DevR response regulator in Mycobacterium smegmatis and that this inhibitory effect was exerted through phosphorylation of DevR on Thr180 within its DNA-binding domain. Moreover, the purified kinase domain of Mtb PknB significantly phosphorylated RegX3, NarL, KdpE, TrcR, DosR, and MtrA response regulators of Mtb that contain the Thr residues corresponding to Thr180 of DevR in their DNA-binding domains, implying that transcriptional activities of these response regulators might also be inhibited when the kinase domain of PknB is overexpressed.

N-Acetylphytosphingosine Enhances the Radiosensitivity of Lung Cancer Cell Line NCI-H460

  • Han, Youngsoo;Kim, Kisung;Shim, Ji-Young;Park, Changsoe;Song, Jie-Young;Yun, Yeon-Sook
    • Molecules and Cells
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    • v.25 no.2
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    • pp.224-230
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    • 2008
  • Ceramides are well-known second messengers that induce apoptosis in various kinds of cancer cells, and their effects are closely related to radiation sensitivity. Phytoceramides, the yeast counterparts of the mammalian ceramides, are also reported to induce apoptosis. We investigated the effect of a novel ceramide derivative, N-acetylphytosphingosine (NAPS), on the radiosensitivity of NCI-H460 human lung carcinoma cells and its differential cytotoxicity in tumor and normal cells. The combination of NAPS with radiation significantly increased clonogenic cell death and caspase-dependent apoptosis. The combined treatment greatly increased Bax expression and Bid cleavage, but not Bcl-2 expression. However, there was no effect on radiosensitivity and apoptosis in BEAS2B cells, which derive from normal human bronchial epithelium. Cell proliferation and DNA synthesis were significantly inhibited by NAPS in both NCI-H460 and BEAS2B cells, but only the BEAS2B cells recovered by 48h after removal of the NAPS. Furthermore, the NCI-H460 cells underwent more DNA fragmentation than the BEAS2B cells in response to NAPS. Our results indicate that NAPS may be a potential radiosensitizing agent with differential effects on tumor vs. normal cells.

Cytochrome B 유전자의 PCR-RFLP Marker에 의한 식육자원의 축종 판별

  • Sin, Seong-Cheol;Choe, Eun-Ju;Heo, Yeon-Beom;Baek, Myeong-Gi;Gwon, Su-Yeon;Kim, Bo-Hyeon;Jeong, Gu-Yong;Jeong, Ui-Ryong
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2005.05a
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    • pp.195-198
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    • 2005
  • 본 연구는 cytochrome B 유전자의 PCR-RFLP 분석기법을 이용하여 다양한 식육자원 및 각종 가공 육제품의 원료육에 대한 정확하고 재현성 높은 축종 및 육종 감별기술을 개발하기 위하여 수행되었다. 국내에서 유통되고 있는 7종류 축종(닭, 양, 돼지, 소, 사슴, 말, 염소)의 육류로부터 cytochrome B 유전자의 특정 염기서열을 포함하는 primer를 설계 제작하여 PCR-RFLP 분석을 실시하였다. 각 축종의 근육조직으로부터 genomic DNA를 추출하고 PCR 증폭 반응을 수행한 후 얻어진 PCR 증폭산물(359 bp)을 두 종류의 제한효소(Hae Ш 및 Hinf I)로 각각 절단한 결과 축종 간 그리고 제한효소 간에 명확한 차이를 보이는 종 특이적인 PCR-RFLP marker를 검출하였다. 따라서, 본 연구에서 개발한 cytochrome B 유전자의 종 특이적 PCR-RFLP marker는 각종 식육 및 가공 육제품의 육종 및 축종 판별에 매우 유용한 DNA marker로 이용될 수 있을 것이다.

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Identification of Differentially Expressed Genes by TCDD in Human Bronchial Cells: Toxicogenomic Markers for Dioxin Exposure

  • Park, Chung-Mu;Jin, Kyong-Suk;Lee, Yong-Woo
    • Biomedical Science Letters
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    • v.18 no.1
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    • pp.1-9
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    • 2012
  • Differentially expressed genes by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were identified in order to evaluate them as dioxin-sensitive markers and crucial signaling molecules to understand dioxin-induced toxic mechanisms in human bronchial cells. Gene expression profiling was analyzed by cDNA microarray and ten genes were selected for further study. They were cytochrome P450, family 1, subfamily B, polypeptide 1 (CYP1B1), S100 calcium binding protein A8 (calgranulin A), S100 calcium binding protein A9 (calgranulin B), aldehyde dehydrogenase 1 family, member A3 (ALDH6) and peroxiredoxin 5 (PRDX5) in up-regulated group. Among them, CYP1B1 was used as a hallmark for dioxin and sharply increased by TCDD exposure. Down-regulated genes were IK cytokine, interferon-induced protein with tetratricopeptide repeats 1 (IFIT1), nuclease sensitive element binding protein 1 (NSEP1), protein tyrosine phosphatase type VI A, member 1 (PTP4A1), ras oncogene family 32 (RAB32). Although up-regulated 4 genes in microarray were coincided with northern hybridization, down-regulated 5 genes showed U-shaped expression pattern which is sharply decreased at lower doses and gradually increased at higher doses. These results introduce some of TCDD-responsive genes can be sensitive markers against TCDD exposure and used as signaling cues to understand toxicity initiated by TCDD inhalation in pulmonary tissues.

Additional Records of the Hydrothermal Vent Scale Worm Branchinotogluma segonzaci (Polynoidae: Lepidonotopodinae) from the North Fiji Basin and Tonga Arc

  • Lee, Won-Kyung;Lee, Geon Hyeok;Ju, Se-Jong;Kim, Se-Joo
    • Animal Systematics, Evolution and Diversity
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    • v.37 no.4
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    • pp.273-279
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    • 2021
  • Branchinotogluma segonzaci (Miura and Desbruyères, 1995) occurs in hydrothermal vent fields of the southwestern Pacific Ocean. We morphologically compared B. segonzaci from the North Fiji Basin with the original description from the Lau Basin and a subsequent study of specimens from the Manus Basin. The main characteristics of all B. segonzaci populations were similar having 21 segments, 10 pairs of elytra, cylindrical-shaped anterior lobes, and ventral papillae on segment 12 and ventral lamellae on segments 13-17 in males. However, the specimens from the North Fiji Basin had rounded to sub-renifrom elytra rather than oval in the original description. Additionally, we newly obtained 11 cytochrome oxidase subunit I (COI) DNA barcodes from the North Fiji Basin and Tonga Arc populations and compared them with known COI DNA barcodes of Branchinotogluma species. Thirteen sequences of B. segonzaci showed 0.0-1.07% intraspecific variation and formed two clades in the COI neighbor-joining tree, whereas the interspecific variation among Branchinotogluma species was 8.19-22.4%. The results of this study contribute to biogeographic studies of B. segonzaci and the evolution of polynoid scale worms in chemosynthesis-based ecosystems.

A Fluorescent Recombinase Aided Amplification Assay for Detection of Babesia microti

  • Lin, Hong;Zhao, Song;Ye, Yuying;Shao, Lei;Jiang, Nizhen;Yang, Kun
    • Parasites, Hosts and Diseases
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    • v.60 no.3
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    • pp.201-205
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    • 2022
  • Babesia microti is one of the most common causative agents of babesiosis. A sensitive and rapid detection is necessary for screening potentially infected individuals. In this study, B. microti cytochrome c oxidase subunit I (cox1) was selected as the target gene, multiple primers were designed, and optimized by a recombinase-aided amplification (RAA) assay. The optimal primers and probe were labeled with fluorescein. The sensitivity of fluorescent RAA (fRAA) was evaluated using gradient diluents of the cox1 recombinant plasmid and genomic DNA extracted from whole blood of B. microti infected mice. The specificity of fRAA was assessed by other transfusion transmitted parasites. The analytical sensitivity of the fRAA assay was 10 copies of recombinant plasmid per reaction and 10 fg/µl B. microti genomic DNA. No cross-reaction with any other blood-transmitted parasites was observed. Our results demonstrated that the fRAA assay would be rapid, sensitive, and specific for the detection of B. microti.

Genetic Variation and Identification of RAPD Markers from Some Garlic Cultivars in Korea and Mongolia (한국과 몽고 일부 재배마늘의 유전적 변이와 재배종 특이적 RAPD 마커의 탐색)

  • Bae, Seong-Kuk;Jung, Eun-A;Kwon, Soon-Tae
    • Korean Journal of Plant Resources
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    • v.23 no.5
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    • pp.458-464
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    • 2010
  • Twelve garlic cultivars collected from Korea and Mongolia were evaluated genetic similarity and diversity by RAPD method using oligo-nucleotide random primers. Genomic DNA isolated from twelve garlic cultivars were amplified by polymerase chain reaction using 143 primers, and 55 primers showed polymorphic DNA bands. Among a total of 187 bands amplified by 55 primers, 128 polymorphic bands were subjected to analysis for genetic relationship of garlic cultivars. Garlic cultivars were classified into three groups, such as group-I corresponded to Euiseong, Seosan, Samchuk and Yecheon-A, Yechun-B, Euiseong-norang, Jeongsun, Namdo, Yookback and Danyang cultivars, and group-II to Mongolia and group-III to Daeseo cultivars. Thirty DNA bands showing unique specificity to the specific cultivars are likely to be useful for identification of garlic local cultivars as DNA markers.

Cloning and Expression of Human Clotting Factor 9 cDNA un Escherichia coli (인체 혈액응고 9인자 cDNA cloning 및 Escherichia coli 에서의 발현)

  • Young Won Lee;Hyang Suk Hur;Myoung Hee Kim
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.231-240
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    • 1996
  • Human blood clotting (coagulation) factor 9 cDNA which codes for 461 amino acid has been cloned by screening human fetal liver cDNA library using PCR. This 1.4 kb cDNA spanning from the ATG initiation codon to the TAA termination codon was cloned into bacterial .expression vector pGEX-2T, generating pGEX-F9 plasmid. The plasmid pGEX-F9 expresses about 73 kDa GST (Glutathione S-transferase)-Factor 9 fusion protein when introduced into E. coli. Western blot analysis using polyclonal antibody raised against human factor 9 confirmed this fusion protein contains factor 9 protein. The level of GST-factor 9 expression was about 20% of total protein and the purification of fusion protein was efficiently achieved by using GST agarose bead based on one step purification protocol.

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The production and immunostimulatory activity of double-stranded CpG-DNA

  • Park, Byoung-Kwon;Kim, Dong-Bum;Rhee, Jae-Won;Kim, Min-Soo;Seok, Hyun-Jeong;Choi, Soo-Young;Park, Jin-Seu;Lee, Young-Hee;Kwon, Hyung-Joo
    • BMB Reports
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    • v.43 no.3
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    • pp.164-169
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    • 2010
  • CpG-DNA, which contains unmethylated CpG dinucleotides in the context of specific sequences, has remarkable and diverse immunological effects, including induction of proinflammatory cytokine expression and regulation of the Th1/Th2 immune response. Here, we examined the immunostimulatory activities of double-stranded (ds) CpG-DNA in the human B cell line RPMI8226. To investigate whether dsCpG-DNA stimulates immune cells, we constructed a plasmid containing repeated dsCpG-DNA and produced dsCpG-DNA by PCR amplification and EcoR I digestion. PCR-amplified dsCpG-DNA alone did not have immmunostimulatory activity. However, dsCpGDNA encapsulated with lipofectin induced IL-8 promoter activation, HLA-DRA expression, and IL-8 expression in a CG sequence-independent manner. The effects of encapsulated dsCpGDNA were independent of minor endotoxin contamination. These findings suggest the potential use of dsCpG-DNA as a therapy for immune response regulation.

A Possible Target for the Heat Inactivation of SCK Tumor Cells

  • 강만식;정주영
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.305-313
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    • 1989
  • The present investigation aims at inquiring into a possible target for the heat inactivation of SCK tumor cells by comparing the kinetics of cell survival, rate of protein synthesis, and DNA polymerase activity in the presence of heat protector or heat sensitirer. A possible conclusion to be drawn from the present experiment is that there is no direct correlation between cell death and decrease in the rate of protein synthesis, but that the loss of DNA polvmerase $\beta$ activity correlates quite well with cell inactivation. Thus, protein degrada-tion and/or abnormal protein synthesis causes cell inactivation innireuv, possibly by altering the cellular environment which in turn affects the DNA polymerase $\beta$ activity. Accordingly, further studies, dealing with the correlation between changes in the cellular environment and DNA polymerase $\beta$ activity, are needed to set insight into a possible target for the heat inactivation of cells. 본 연구는 열보호제 또는 열증감제의 존재하에서 세포 생존곡선, 단백질 합성률, DNA 중합효소 $\beta$의 활성변화를 비교 검토함으로써 SCK 종양세포가 열에 의해서 불활성화될 때의 표적이 무엇인지를 밝혀보기 위해서 수행되었다. 본 실험의 결과로 추정하건대 열에 의한 세포치사는 단백질 합성률의 변화와는 직접적인 연관성이 없으나, DNA 중합효소 $\beta$의 활성도와는 밀접한 연관성이 있음을 알 수 있다. 즉, 단백질의 분해 또는 비정상적인 단백질의 합성이 세포의 환경을 변화시키고 이것이 DNA 중합효소 $\beta$의 활성에 영향을 미침으로써 간접적으로 세포의 치사를 초래할 것으로 짐작할 수 있다. 따라서, 세포의 열불화성화의 표적을 좀더 분명히 밝히기 위해서는 세포의 환경변화와 DNA 중합효소 $\beta$의 활성과의 관계를 추구하는 연구가 수행되어야 할 것으로 사료된다.

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