• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.593-598
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• 2004

The arsenical resistance (ars) operon was cloned from a 67-kilobase pair (kb) plasmid, which was previously shown to be responsible for arsenic salts resistance in K. oxytoca D12. When plasmid pAE48, carrying the ars operon, was transformed into E. coli, transformed cells displayed enhanced survival in the presence of 4 mM arsenite, 50 mM arsenate, or 0.4 mM antimonite. The nucleotide sequence of the 5.6-kb fragment encoding arsenical resistance revealed five open reading frames (ORFs), which were predicted to encode polypeptides of 12.8 (arsR), 13.4 (arsD), 62.6 (arsA), 45 (arsB), and 16.7 (arse) kilodaltons (kDa). Each ORF was preceded by a ribosome binding site. A putative promoter-like sequence was identified upstream of arsR, and a possible termination site was found downstream of arsC. When the deduced amino acid sequences of the K. oxytoca Dl2 Ars proteins were compared with the amino acid sequences of the E. coli R773 Ars proteins, a significant amino acid similarity was observed (87.9％ for ArsR, 89.2％ for ArsD, 83.2％ for ArsA, 92.6％ for ArsB, and 91.3％ for ArsC), suggesting an evolutionary relationship of the ars genes of E. coli plasmid R773 and K. oxytoca Dl2.

• Journal of Microbiology and Biotechnology
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• 제19권3호
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• pp.217-228
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• 2009

Mobile genetic segments, or transposons, are also referred to as jumping genes as they can shift from one position in the genome to another, thus inducing a chromosomal mutation. According to the target site-specificity of the transposon during a transposition event, the result is either the insertion of a gene of interest at a specific chromosomal site, or the creation of knockout mutants. The former situation includes the integration of conjugative transposons via site-specific recombination, several transposons preferring a target site of a conserved AT-rich sequence, and Tn7 being site-specifically inserted at attTn7, the downstream of the essential glmS gene. The latter situation is exploited for random mutagenesis in many prokaryotes, including IS (insertion sequence) elements, mariner, Mu, Tn3 derivatives (Tn4430 and Tn917), Tn5, modified Tn7, Tn10, Tn552, and Ty1, enabling a variety of genetic manipulations. Randomly inserted transposons have been previously employed for a variety of applications such as genetic footprinting, gene transcriptional and translational fusion, signature-tagged mutagenesis (STM), DNA or cDNA sequencing, transposon site hybridization (TraSH), and scanning linker mutagenesis (SLM). Therefore, transposon-mediated genetic engineering is a valuable discipline for the study of bacterial physiology and pathogenesis in living hosts.

• 미생물학회지
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• 제30권6호
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• pp.472-478
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• 1992

HTLV-1 이 pol 유전자산물을 합성하기 위해서는 genome-size mRNA 를 번역해 나아가는 ribosome 이 -1 방향으로 두차례 frame 을 바꾸어야 한다. 우리는 단 한차계의 frameshifting 만으로도 많은 양의 Gag-Pro-Pol polyprotein 의 합성어 가능하도록 gag 와 pro 유전자의 frame 을 연결시킨 mutant RNA 를 제작하였다. 이 돌연변이를 이용하여 ribosome 의 shift site 하류영역내에 형성이 예상되는 RNA 의 이차구조 또는 삼차 구조가 -1 frameshifting 을 결정하는 인자로서 작용하는지의 여부를 조사하였다. 결손변이주의를 해석한 결과 pro-pol 중첩영역에서 효율적으로 frameshifting 이 일어나기 위해서는 stem-loop 가 필수적으로 형성되어야 하지만 pseudoknot 의 형성은 그다지 중요하지 않다는 사실을 알았다.

• Biomolecules & Therapeutics
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• 제24권2호
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• pp.191-198
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• 2016

The vitamin D receptor (VDR) is a member of the nuclear receptor (NR) superfamily. The VDR binds to active vitamin $D_3$ metabolites, which stimulates downstream transduction signaling involved in various physiological activities such as calcium homeostasis, bone mineralization, and cell differentiation. Quercetin is a widely distributed flavonoid in nature that is known to enhance transactivation of VDR target genes. However, the detailed molecular mechanism underlying VDR activation by quercetin is not well understood. We first demonstrated the interaction between quercetin and the VDR at the molecular level by using fluorescence quenching and saturation transfer difference (STD) NMR experiments. The dissociation constant ($K_d$) of quercetin and the VDR was $21.15{\pm}4.31{\mu}M$, and the mapping of quercetin subsites for VDR binding was performed using STD-NMR. The binding mode of quercetin was investigated by a docking study combined with molecular dynamics (MD) simulation. Quercetin might serve as a scaffold for the development of VDR modulators with selective biological activities.

• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3505-3508
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• 2015

Background: Ras genes are thought to play an important role in human cancer since they have been found to be activated frequently in several types of tumors including breast cancer, where the overall incidence of K-RAS oncogene activation is 0-10%. Evaluation of K-RAS gene not only for mutational frequency but also for mutation types in this downstream signaling gene pathway is necessary to determine the mechanisms of action. The present study was conducted to test the hypothesis that K-RAS activation is involved in breast cancer risk of south Indian population. Materials and Methods: A total of 70 paired pathologically confirmed tumor and non-tumor tissues from the same breast cancer patients were analysed for most common K-RAS mutations of codon 12,13 and 61 by polymerase chain reaction followed by restriction digestion and direct nucleotide sequencing method. Results: We found that a high rate of homozygous and heterozygous mutations of codon 12, but not codon 13 and 61, may influence the invasive ductal carcinoma of breast risk in this study. Conclusions: Our study indicated that only codon 12 may be involved in initiating breast carcinogenesis in India.

• The Korean Journal of Physiology and Pharmacology
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• 제14권5호
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• pp.331-336
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• 2010

In rapidly growing tumors, hypoxia commonly develops due to the imbalance between $O_2$ consumption and supply. Hypoxia Inducible Factor (HIF)-$1{\alpha}$ is a transcription factor responsible for tumor growth and angiogenesis in the hypoxic microenvironment; thus, its inhibition is regarded as a promising strategy for cancer therapy. Given that CamKII or PARP inhibitors are emerging anticancer agents, we investigated if they have the potential to be developed as new HIF-$1{\alpha}$-targeting drugs. When treating various cancer cells with the inhibitors, we found that a CamKII inhibitor, KN-62, effectively suppressed HIF-$1{\alpha}$ specifically in hepatoma cells. To examine the effect of KN-62 on HIF-$1{\alpha}$-driven gene expression, we analyzed the EPO-enhancer reporter activity and mRNA levels of HIF-$1{\alpha}$ downstream genes, such as EPO, LOX and CA9. Both the reporter activity and the mRNA expression were repressed by KN-62. We also found that KN-62 suppressed HIF-$1{\alpha}$ by impairing synthesis of HIF-$1{\alpha}$ protein. Based on these results, we propose that KN-62 is a candidate as a HIF-$1{\alpha}$-targeting anticancer agent.

• The Plant Pathology Journal
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• 제36권2호
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• pp.121-132
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• 2020

β-Aminobutyric acid (BABA) has consistently been reported to enhance plant immunity. However, the specific mechanisms and downstream components that mediate this resistance are not yet agreed upon. Nitric oxide (NO) is an important signal molecule involved in a diverse range of physiological processes, and whether NO is involved in BABA-induced resistance is interesting. In this study, treatment with BABA significantly increased NO accumulation and reduced the sensitivity to Botrytis cinerea in tomato plants. BABA treatment reduced physical signs of infection and increased both the transcription of key defense marker genes and the activity of defensive enzymes. Interestingly, compared to treatment with BABA alone, treatment with BABA plus cPTIO (NO specific scavenger) not only significantly reduced NO accumulation, but also increased disease incidence and lesion area. These results suggest that NO accumulation plays an important role in BABA-induced resistance against B. cinerea in tomato plants.

• 대한한방내과학회지
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• 제24권1호
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• pp.134-143
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• 2003

Objectives : We performed this study to understand the molecular basis of the antitumor effect of Saussurea lappa, Pharbitis nil, Plantago asiatica and Taraxacum mongolicum, which have been used for cancer treatment in Korean traditional medicine. Design: We analyzed, the effect of these medicinal herbs on proliferation and apoptosis of tumor cells and its association with gene expression, We performed semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) analysis of cell cycle- and apoptosis-related genes using a gastric cancer cell line AGS. Results : Cell counting assay and $[^3H]thymidine$ uptake analysis showed that Saussurea lappa and Pharbitis nil strongly inhibit cell proliferation of AGS in a dose-dependent manner. Interestingly, gene espression assay revealed that mRNA espression levels of c-Jun, c-Fos, c-Myc, and Cyclin D1 were markedly decreased by Saussurea lappa and Pharbitis nil. Furthermore, Saussurea lappa was identified to activate expression of the p53 tumor suppressor and its downstream effector $p21^{Wafl}$, which leads to $G_1$ cell cycle arrest and apoptosis. These observations suggest that the anticancer effect of Saussurea lappa and Pharbitis nil might be associated with their regulatory capability of tumor-related gene expression.

• Molecules and Cells
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• 제28권4호
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• pp.365-368
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• 2009

Toll-like receptors (TLRs) play an important role in host defense by sensing invading microbial pathogens and initiating innate immune responses. The stimulation of TLRs by microbial components triggers the activation of myeloid differential factor 88 (MyD88)- and toll-interleukin-1 receptor domain-containing adapter inducing interferon-${\beta}$ (TRIF)-dependent downstream signaling pathways. Isoliquiritigenin (ILG), an active ingredient of Licorice, has been used for centuries to treat many chronic diseases. ILG inhibits the MyD88-dependent pathway by inhibiting the activity of inhibitor-${\kappa}B$ kinase. However, it is not known whether ILG inhibits the TRIF-dependent pathway. To evaluate the therapeutic potential of ILG, we examined its effect on signal transduction via the TRIF-dependent pathway of TLRs induced by several agonists. ILG inhibited nuclear factor-${\kappa}B$ and interferon regulatory factor 3 activation induced by lipopolysaccharide or polyinosinic-polycytidylic acid. ILG inhibited the lipopolysaccharide-induced phosphorylation of interferon regulatory factor 3 as well as interferon-inducible genes such as interferon inducible protein-10, and regulated activation of normal T-cell expressed and secreted (RANTES). These results suggest that ILG can modulate TRIF-dependent signaling pathways of TLRs, leading to decreased inflammatory gene expression.

• Genomics & Informatics
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• 제10권3호
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• pp.153-166
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• 2012

A variety of ligands differ in their capacity to bind the receptor, elicit gene expression, and modulate physiological responses. Such receptors include Toll-like receptors (TLRs), which recognize various patterns of pathogens and lead to primary innate immune activation against invaders, and G-protein coupled receptors (GPCRs), whose interaction with their cognate ligands activates heterotrimeric G proteins and regulates specific downstream effectors, including immuno-stimulating molecules. Once TLRs are activated, they lead to the expression of hundreds of genes together and bridge the arm of innate and adaptive immune responses. We characterized the gene expression profile of Toll-like receptor 4 (TLR4) in RAW 264.7 cells when it bound with its ligand, 2-keto-3-deoxyoctonate (KDO), the active part of lipopolysaccharide. In addition, to determine the network communications among the TLR, Janus kinase (JAK)/signal transducer and activator of transcription (STAT), and GPCR, we tested RAW 264.7 cells with KDO, interferon-${\beta}$, or cAMP analog 8-Br. The ligands were also administered as a pair of double and triple combinations.