• 대한수의학회지
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• 제52권2호
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• pp.99-104
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• 2012

The present study was performed to evaluate the relationship between the neurotoxicity of acrylamide and the differential gene expression pattern in mice. Both locomotor test and rota-rod test showed that the group treated with higher than 30 mg/kg/day of acrylamide caused impaired motor activity in mice. Based on cDNA microarray analysis of mouse brain, myelin basic protein gene, kinesin family member 5B gene, and fibroblast growth factor (FGF) 1 and its receptor genes were down-regulated by acrylamide. The genes are known to be essential for neurofilament synthesis, axonal transport, and neuroprotection, respectively. Interestingly, both FGF 1 and its receptor genes were down-regulated. Genes involved in nucleic acid binding such as AU RNA binding protein/enoyl-coA hydratase, translation initiation factor (TIF) 2 alpha kinase 4, activating transcription factor 2, and U2AF 1 related sequence 1 genes were down-regulated. More interesting finding was that genes of both catalytic and regulatory subunit of protein phosphatases which are important for signal transduction pathways were down-regulated. Here, we propose that acrylamide induces neurotoxicity by regulation of genes associated with neurofilament synthesis, axonal transport, neuro-protection, and signal transduction pathways.

• Journal of Acupuncture Research
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• 제22권2호
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• pp.133-139
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• 2005

Cobrotoxin, a venom of Vipera lebetina turanica, is a group of basic peptidescomposed of 233 amino acids with six disulfide bonds formed by twelve cysteins. NF-kB is activated by subsequent release of inhibitory IkB and translocation of p50. Since sulfhydryl group is present in kinase domain of p50 subunit of NF-kB, cobrotoxin could modify NF-kB activity by protein-protein interaction. We therefore examined effect of cobrotoxin on NF-kB activities in lipopolysaccharide (LPS) and sodium nitroprusside (SNP)-stimulated Raw 264.7 mouse macrophages. Cobrotoxin suppressed the LPS and SNP-induced release of IkB and p50 translocation resulted in inhibition of DNA binding activity of NF-kB. Inhibition of NF-kB resulted in reduction of the LPS and SNP-induced production of inflammatory mediators NO and PGE2 generation. The inhibitory effect of cobrotoxin on the NF-kB activity were blocked by addition of reducing agents dithiothreitol and glutathione. These results demonstrate that cobrotoxin inhibits activation of NF-kB, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-kB signal pathway.

• 한국균학회지
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• 제49권4호
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• pp.545-553
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• 2021

In 2021, seven yeast strains were isolated from earthworm (Eisenia andrei) gut samples collected from the Nanji Water Regeneration Center in Goyang City, Gyeonggi Province, Korea. A total of seven yeasts were isolated, of which three strains have not been previously reported in Korea. To identify the yeasts, pairwise sequence comparisons of large subunit (LSU) rDNA sequences were performed using the basic local alignment search tool (BLAST). Assimilation test and cell morphology analysis were performed using the API 20C AUX kit and phase contrast microscope, respectively. Five of the seven strains were assigned to the genus Candida of the order Saccharomycetales of the class Saccharomycetes, and two to the genus Apiotrichum of the order Trichosporonales of the class Tremellomycetes. The yeast strain Candida sojae E2 belongs to the family Debaryomycetaceae, and Apiotrichum laibachii E8 and A. laibachii E9 belong to the family Trichosporonaceae. All strains were cultured in yeast mold agar for three days and showed different colony forms. C. sojae E2 was round and entire shaped, while A. laibachii E8 and A. laibachii E9 was round and convex shaped. This study focuses on the description of the three yeast strains that have not been officially reported in Korea.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.959-966
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• 2004

Twenty-three cultures of harmful algal bloom (HAB)-(causing dinoflagellates were isolated from the coastal waters of Korea. For each of the 14 morphospecies, the nuclearencoded small subunit (SSU) rDNA was analyzed to determine the phylogenetic relatedness of the species. Despite temporal and spatial isolation, 3-4 clonal cultures of Alexandrium catenella, Cochlodinium polykrikoides, and Gymnodinium catenatum had 100% identical SSU rDNA sequences. In contrast, heterogeneities in the SSU rDNA sequences were observed in Akashiwo sanguinea and Lingulodinium polyedrum strains. Extreme sequence polymorphism was shown within the SSU rRNA genes of an Al. tamarense clonal culture. A homology search in GenBank revealed that 11 dinoflagellate species were located in clusters corresponding to their morphological classification. The SSU rDNA sequences of C. polykrikoides, Gyrodinium instriatum, and Pheopolykrikos hartmannii, which were determined for the first time in this study, showed the following phylogenetic relationships: C. polykrikoides formed an independent branch separated from other dinoflagellates; Gyr. instriatum was placed in a monophyletic group with Gyr. dorsum and Gyr. uncatenum; and Ph. hartmanii, which forms a distinct two-celled pseudocolony, belonged to Gymnodinium sensu Hansen and Moestrup.

• 미생물학회지
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• 제51권4호
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• pp.435-439
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• 2015

THO/TREX 복합체는 전사 신장, mRNA 가공 및 방출, 그리고 유전체 안정성에 중요한 역할을 담당한다. 분열효모 Schizosaccharomyces pombe에서 THO/TREX 복합체의 한 구성요소인 THOC5의 이종상동체를 암호화하고 있는 SPBC 577.04 유전자를 찾아, 그것의 기능을 분석하였다. S. pombe thoc5 (spthoc5) 유전자는 생장과 mRNA의 방출에 필수적이지는 않지만, 결실돌연변이는 야생형에 비해 생장 결함을 보였고 $poly(A)^+$ RNA도 핵 안에 약간 축적되는 현상을 보였다. 또한 정상적인 기능을 가진 spThoc5-GFP 단백질은 주로 핵안에 존재하였다. Co-immunoprecipitation 분석에서 진화적으로 잘 보존된THO/TREX 복합체의 주요 구성인자인 Hpr1(THOC1)는 또 다른 구성인자인 Tho2 (THOC2) 뿐만 아니라 spThoc5와도 상호작용을 하였다. 이와 같은 결과들은 S. pombe의 Thoc5 상동체도 THO/TREX 복합체의 구성인자로 mRNA 방출에 관여하고 있음을 시사한다.

• 미생물학회지
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• 제48권2호
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• pp.171-174
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• 2012

THOC1/Hpr1는 mRNA가 전사되는 동안 mRNP의 포장과 mRNA 방출에 관여하는 진화적으로 잘 보존된 THO 복합체의 한 소단위이다. 분열효모 Schizosaccharomyces pombe에서도 THOC1/Hpr1과 유사한 단백질을 암호화하는 유전자(sphpr1로 명명)를 찾아 그 특성을 조사하였다. 이배체 S. pombe 균주에 하나의 sphpr1 유전자만을 결실시킨 후 4분체 분석을 수행한 결과, 이 유전자는 생장에 필수적이었다. 티아민에 의해 발현이 조절되는 강력한 nmt1 프러모터를 이용하여 sphpr1를 과발현시키더라도 세포의 생장과 mRNA 방출에는 전혀 영향이 없었다. 하지만, sphpr1의 발현을 억제하면 생장이 억제되었으며 poly$(A)^+$ RNA가 핵 안에 축적되었다. 이와 같은 결과들은 sphpr1 유전자가 생장과 mRNA의 핵에서 세포질로의 방출에 관여하고 있음을 시사한다.

• Journal of Microbiology
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• 제45권5호
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• pp.418-421
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• 2007

The nucleotide at position 791(G791) of E. coli 16S rRNA was previously identified as an invariant residue for ribosomal function. In order to characterize the functional role of G791, base substitutions were introduced at this position, and mutant ribosomes were analyzed with regard to their protein synthesis ability, via the use of a specialized ribosome system. These ribosomal RNA mutations attenuated the ability of ribosomes to conduct protein synthesis by more than 65%. A transition mutation (G to A) exerted a moderate effect on ribosomal function, whereas a transversion mutation (G to C or U) resulted in a loss of protein synthesis ability of more than 90%. The sucrose gradient profiles of ribosomes and primer extension analysis showed that the loss of protein-synthesis ability of mutant ribosomes harboring a base substitution from G to U at position 791 stems partially from its inability to form 70S ribosomes. These findings show the involvement of the nucleotide at position 791 in the association of ribosomal subunits and protein synthesis steps after 70S formation, as well as the possibility of using 16S rRNA mutated at position 791 for the selection of second-site revertants in order to identify ligands that interact with G791 in protein synthesis.

• IMMUNE NETWORK
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• 제3권1호
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• pp.38-46
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• 2003

Background: Platelet-activating factor (PAF) induces nuclear factor $(NF)-{\kappa}B$ activation and angiogenesis and increases tumor growth and pulmonary tumor metastasis in vivo. The role of $NF-{\kappa}B$ activation in PAF-induced angiogenesis in a mouse model of Matrigel implantation, and in PAF-mediated pulmonary tumor metastasis were investigated. Methods: Angiogenesis using Matrigel and experimental pulmonary tumor metastasis were tested in a mouse model. Electrophoretic mobility shift assay was done for the assessment of $NF-{\kappa}B$ translocation to the nucleus. Expression of angiogenic factors, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\alpha}$, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were tested by RT-PCR and ELISA. Results: PAF induced a dose- and time-dependent angiogenic response. PAF-induced angiogenesis was significantly blocked by PAF antagonist, CV6209, and inhibitors of $NF-{\kappa}B$ expression or action, including antisense oligonucleotides to p65 subunit of $NF-{\kappa}B$ (p65 AS) and antioxidants such as ${\alpha}$-tocopherol and N-acetyl-L-cysteine. In vitro, PAF activated the transcription factor, $NF-{\kappa}B$ and induced mRNA expression of $TNF-{\alpha}$, $IL-1{\alpha}$, bFGF, VEGF, and its receptor, KDR. The PAF-induced expression of the above mentioned factors was inhibited by p65 AS or antioxidants. Also, protein synthesis of VEGF was increased by PAF and inhibited by p65 AS or antioxidants. The angiogenic effect of PAF was blocked when anti-VEGF antibodies was treated or antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF was co-administrated, but not by antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF each alone. PAF-augmented pulmonary tumor metastasis was inhibited by p65 AS or antioxidants. Conclusion: These data indicate that PAF increases angiogenesis and pulmonary tumor metastasis through $NF-{\kappa}B$ activation and expression of $NF-{\kappa}B$-dependent angiogenic factors.

• 미생물학회지
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• 제50권2호
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• pp.164-168
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• 2014

북극지역은 지구온난화로 인하여 생태계에 큰 영향을 받고 있다. 본 연구는 북극 Svalbard 지역에서 육지 빙하의 해빙(ice melt)의 영향을 받는 해양퇴적층에서 질산화 과정에 핵심역할을 하는 고세균의 질산화유전자(ammonia monooxygenase, AMO)의 공간적 분포의 변화를 조사하였다. 해빙으로 인한 퇴적물 퇴적속도와 고세균 AMO의 alpha subunit를 코딩하는 amoA 유전자와의 관계를 클론라이브러리 분석을 통하여 분석하였다. 육지와 근접하여 퇴적속도가 가장 빠른 정점(188)에서 고세균 amoA 유전자의 다양성이 육지에서 비교적 먼 지역의 정점(176과 184)에 비해 현저히 낮음을 알 수 있었다. 3 정점의 고세균 amoA 유전자 클론들의 평균 아미노산 서열의 상동성은 94%(염기서열 91%)로 나타났다. 176과 184 정점에서 분석된 고세균 amoA 유전자 클론들 중 약 45%가 Nitosopumilus clade와 근연관계에 있는 반면, 188 지역의 경우 낮은 염농도에서 발견되는 Nitrosoarchaeaum clade와 근연관계에 있는 클론들이 발견되었다. 토양 고세균유래 amoA 유전자는 육지에 근접하여 해빙에 의한 영향을 가장 많이 받는 188정점에서만 발견이 되었다. 본 연구를 통하여, 해빙으로 인하여 육지로부터 운반되는 퇴적물의 량이 증가함에 따라, 해양퇴적층의 질소순환관련 미생물 군집에 변화가 유발되는 것으로 추정되며, 고세균의 amoA 유전자가 해양퇴적층 질소순환생태계 변화의 지표로 이용될 수 있음을 알 수 있었다.

• 생명과학회지
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• 제25권9호
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• pp.1007-1013
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• 2015

원핵생물체의 생명유지에 중요한 역할을 담당하는 유전자들을 밝히기 위해 미생물 유전체들 사이의 공통적 유전자를 파악하는 COG 알고리즘을 이용하였다. 원핵생물 711종 모두에 보존적인 것은 COG0080 (Ribosomal protein L11) 1개였다. 708종 이상의 원핵생물에 보존적인 22개의 ortholog 중 전사관련 2개, tRNA synthetase 관련4개, ribosamal large subunit 8개, ribosomal small subunit 7개였다. 700종 이상의 원핵생물에 보존적인 COG는 58개였다. 이중 리보좀을 구성하는 소단위체 등 번역 관련 COG가 50개(86.2%), 전사관련 COG가 4개(6.9%)로 나타나 생명현상에서의 단백질의 중요성을 알 수 있었다. 58개의 COG 중 보존성은 COG0060 (Isoleucyl tRNA synthetase)이 가장 높았고 COG0143 (Methionyl tRNA synthetase)이 가장 낮았다. 문(phylum)과 강(class) 수준에서 보존적 유전자들의 평균과 분산으로 유전체 분석을 수행한 결과 변이가 큰 고세균은 진정세균과 구분되었으며 편차는 일부 진정세균이 고세균보다 컸다. 보존적 유전자를 탐색하는 본 연구의 기법은 기초과학 연구와 함께 항균제 개발과 항암요법 개발 등에도 유용할 것이다.