• Journal of Microbiology and Biotechnology
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• 제26권1호
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• pp.160-170
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• 2016

The increasing spread of antibiotic-resistant pathogens has raised the interest in alternative antimicrobial treatments. In our study, the functionally active gram-negative bacterium bacteriophage CP933 endolysin was produced in Nicotiana benthamiana plants by a combination of transient expression and vacuole targeting strategies, and its antimicrobial activity was investigated. Expression of the cp933 gene in E. coli led to growth inhibition and lysis of the host cells or production of trace amounts of CP933. Cytoplasmic expression of the cp933 gene in plants using Potato virus X-based transient expression vectors (pP2C2S and pGR107) resulted in death of the apical portion of experimental plants. To protect plants against the toxic effects of the CP933 protein, the cp933 coding region was fused at its Nterminus to an N-terminal signal peptide from the potato proteinase inhibitor I to direct CP933 to the delta-type vacuoles. Plants producing the CP933 fusion protein did not exhibit the severe toxic effects seen with the unfused protein and the level of expression was 0.16 mg/g of plant tissue. Antimicrobial assays revealed that, in contrast to gram-negative bacterium E. coli (BL21(DE3)), the gram-positive plant pathogenic bacterium Clavibacter michiganensis was more susceptible to the plant-produced CP933, showing 18% growth inhibition. The results of our experiments demonstrate that the combination of transient expression and protein targeting to the delta vacuoles is a promising approach to produce functionally active proteins that exhibit toxicity when expressed in plant cells.

• 한국발생생물학회지:발생과생식
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• 제17권4호
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• pp.389-397
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• 2013

FGF9/16/20 signaling pathway specify the developmental fates of notochord, mesenchyme, and neural cells in ascidian embryos. Although a conserved Ras/MEK/Erk/Ets pathway is known to be involved in this signaling, the detailed mechanisms of regulation of FGF signaling pathway have remained largely elusive. In this study, we have isolated Hr-Erf, an ascidian orthologue of vertebrate Erf, to elucidate interactions of transcription factors involved in FGF signaling of the ascidian embryo. The Hr-Erf cDNA encompassed 3110 nucleotides including sequence encoded a predicted polypeptide of 760 amino acids. The polypeptide had the Ets DNA-binding domain in its N-terminal region. In adult animals, Hr-Erf mRNA was predominantly detected in muscle, and at lower levels in ganglion, gills, gonad, hepatopancreas, and stomach by quantitative real-time PCR (QPCR) method. During embryogenesis, Hr-Erf mRNA was detected from eggs to early developmental stage embryos, whereas the transcript levels were decreased after neurula stage. Similar to the QPCR results, maternal transcripts of Hr-Erf was detected in the fertilized eggs by whole-mount in situ hybridization. Maternal mRNA of Hr-Erf was gradually lost from the neurula stage. Zygotic expression of Hr-Erf started in most blastomeres at the 8-cell stage. At gastrula stage, Hr-Erf was specifically expressed in the precursor cells of brain and mesenchyme. When MEK inhibitor was treated, embryos resulted in loss of Hr-Erf expression in mesenchyme cells, and in excess of Hr-Erf in a-line neural cells. These results suggest that zygotic Hr-Erf products are involved in specification of mesenchyme and neural cells.

• 전자공학회논문지S
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• 제35S권11호
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• pp.152-165
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• 1998

영역 기반 부호화 시 윤곽선의 정보량은 부호화 후 전체 정보량의 큰 부분을 차지한다. 본 논문에서는 분할 기반 영상 부호화를 위한 새로운 적응적 윤곽선 부호화 기법을 제안한다. 주변 영역간의 대비를 고려하여 적응적으로 윤곽선을 부호화하는 제안한 기법을 적용하여 주관적 화질의 저하 없이 전체 윤곽선 정보량을 줄일 수 있다. 영상에 형태학적 분수계 알고리즘을 적용하여 영역별로 분할한 후, 윤곽선 세그먼트들을 인접 영역간의 대비에 따라 분류한다. 분류된 윤곽선 세그먼트 중, 낮은 대비 영역 사이의 윤곽선 세그먼트들은 형태학적 저역 통과 필터 등에 의해 크게 압축할 수 있다. 실험을 통해 제안한 윤곽선 부호화 기법은 초저속 환경 하에서 우수한 수행 결과를 나타내었다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권6호
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• pp.3865-3869
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• 2013

Background: Cigarette per day (CPD) use is a key smoking behaviour indicator. It reflects smoking intensity which is directly proportional to the occurrence of tobacco induced cancers. Self reported CPD assessment in surveys may suffer from digit bias and under reporting. Estimates from such surveys could influence the policy decision for tobacco control efforts. In this context, this study aimed at identifying underlying factors of digit bias and its implications for Global Adult Tobacco Surveillance. Materials or Methods: Daily manufactured cigarette users CPD frequencies from Global Adult Tobacco Survey (GATS) - India data were analyzed. Adapted Whipple Index was estimated to assess digit bias and data quality of reported CPD frequency. Digit bias was quantified by considering reporting of '0' or '5' as the terminal digits in the CPD frequency. The factors influencing it were identified by bivariate and logistic regression analysis. Results: The mean and mode of CPD frequency was 6.7 and 10 respectively. Around 14.5%, 15.1% and 15.2% of daily smokers had reported their CPD frequency as 2, 5 and 10 respectively. Modified Whipple index was estimated to be 226.3 indicating poor data quality. Digit bias was observed in 38% of the daily smokers. Heavy smoking, urban residence, North, South, North- East region of India, less than primary, secondary or higher educated and fourth asset index quintile group were significantly associated with digit bias. Discussion: The present study highlighted poor quality of CPD frequency data in the GATS-India survey and need for its improvement. Modeling of digit preference and smoothing of the CPD frequency data is required to improve quality of data. Marketing of 10 cigarette sticks per pack may influence CPD frequency reporting, but this needs further examination. Exploring alternative methods to reduce digit bias in cross sectional surveys should be given priority.

• BMB Reports
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• 제50권12호
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• pp.640-646
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• 2017

Regulatory B cells, also well-known as IL-10-producing B cells, play a role in the suppression of inflammatory responses. However, the epigenetic modulation of regulatory B cells is largely unknown. Recent studies showed that the bromodomain and extra-terminal domain (BET) protein inhibitor JQ1 controls the expression of various genes involving cell proliferation and cell cycle. However, the role of BET proteins on development of regulatory B cells is not reported. In this study, JQ1 potently suppressed IL-10 expression and secretion in murine splenic and peritoneal B cells. While bromodomain-containing protein 4 (BRD4) was associated with $NF-{\kappa}B$ on IL-10 promoter region by LPS stimulation, JQ1 interfered the interaction of BRD4 with $NF-{\kappa}B$ on IL-10 promoter. In summary, BRD4 is essential for toll like receptor 4 (TLR4)-mediated IL-10 expression, suggesting JQ1 could be a potential candidate in regulating IL-10-producing regulatory B cells in cancer.

• The Plant Pathology Journal
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• 제33권1호
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• pp.53-65
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• 2017

Barley yellow dwarf virus (BYDV) belongs to Luteovirus and is limited only at phloem related tissues. An open reading frame (ORF) 4 of BYDV codes for the movement protein (MP) of BYDV gating plasmodesmata (PD) to facilitate virus movement. Like other Luteoviruses, ORF 4 of BYDV is embedded in the ORF3 but expressed from the different reading frame in leaky scanning manner. Although MP is a very important protein for systemic infection of BYDV, there was a little information. In this study, MP was characterized in terms of subcellular localization and programmed cell death (PCD). Gene of MP or its mutant (ΔMP) was expressed by Agroinfiltration method. MP was clearly localized at the nucleus and the PD, but ΔMP which was deleted distal N-terminus of MP showed no localization to PD exhibited the different target with original MP. In addition to PD localization, MP appeared associated with small granules in cytoplasm whereas ΔMP did not. MP associated with PD and small granules induced PCD, but ΔMP showed no association with PD and small granules did not exhibit PCD. Based on this study, the distal N-terminal region within MP is seemingly responsible for the localization of PD and the induction small granules and PCD induction. These results suggest that subcellular localization of BYDV MP may modulate the PCD in Nicotiana benthamiana.

• 한국항만경제학회지
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• 제31권4호
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• pp.121-132
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• 2015

양산 ICD(Inland Container Depot)는 2000년 3월 개장 이후 2005년 133만 TEU를 처리하여 국제물류경쟁력과 부산지역의 컨테이너화물 운송 경쟁력을 높여 부산항 및 우리나라 항만물류산업에서 중요한 역할을 담당하였다. 그러나 2006년 부산항 신항 개장과 신항배후부지의 조성에서 물동량이 급감하여 활성화를 위한 새로운 방안이 모색이 필요한 상황이다. 따라서 본 연구는 AHP(analytic hierarchy process)를 활용하여 평가모형을 설정하고, 양산 ICD 내 입주기업 및 관계자를 대상으로 설문조사를 실시하여 양산 ICD의 활성화 방안을 모색하였다. 연구의 결과, 양산 ICD 활성화 방안으로 입지와 시설의 장점을 고려한 물류시설(공동집배송센터, 물류창고)로의 기능전환이 필요하며, 이용기간 연장, 건폐율 확보를 통해 기존 및 신규 기업활동의 안정성을 보장하여야 할 것이다. 또한 ICD 내 철도역 등 시설을 활용하여 적극적인 철도활성화 정책을 통해 철도택배 등을 활용하여 신규 물동량을 창출하고, 공로 중심의 국가물류체계에서 철송 활성화를 통한 내륙물류기지로서의 그 역할을 제시하였다.

• BMB Reports
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• 제36권5호
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• pp.475-487
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• 2003

A gene that encodes a homologue to baculoviral p74, an envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). A part of the ChfuGV p74 gene was located on an 8.9 kb BamHI subgenomic fragment using different sets of degenerated primers. These were designed using the results of the protein sequencing of a major 74 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1992 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 663 amino acids with a predicted molecular mass of 74,812 Da. Comparative studies revealed the presence of two major conserved regions in the ChfuGV p74 protein. This study also shows that all of the p74 proteins contain two putative transmembrane domains at their C-terminal segments. At the nucleotide sequence level, two late promoter motifs (TAAG and GTAAG) were located upstream of the first ATG of the p74 gene. The gene contained a canonical poly(A) signal, AATAAA, at its 3' non-translated region. A phylogenetic tree for baculoviral p74 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV p74 is related the closest to those of Cydia pomonella granulovirus (CpGV) and Phthorimaea operculella granulovirus (PhopGV).

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
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• pp.278-281
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• 2004

The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.

• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.592-597
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• 2002

The Rkp1/CPC2, a receptor for activated protein kinase C of Schizosaccharomyces pombe, contains seven WD motifs found in the G-protein $\beta$-subunit. A 110-kDa protein was identified to interact with Rkp1/CPC2 by immunoprecipitation and following in vitro binding assay. To examine its kinase activity and binding ability to Rkp1, the $pck1^+$, a PKC homolog of S. pombe, was cloned. Pckl phosphorylated myelin basic protein (MBP) and histone Hl in a phospholipid-dependent and $Ca^{2+}$-independent manner. It was demonstrated that the N-terminal region of Pck1 was responsible for the binding to Rkp1 , thus suggesting that Rkp1 interacted with Pckl to regulate Pckl-mediated signaling in S. pombe.