• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
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• pp.60-61
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• 2004

Metabolic engineering is now a well established discipline, used extensively to determine and execute rational strategies of strain development to improve the performance of micro-organisms employed in industrial fermentations. The basic principle of this approach is that performance of the microbial catalyst should be adequately characterised metabolically so as to clearlyidentify the metabolic network constraints, thereby identifying the most probable targets for genetic engineering and the extent to which improvements can be realistically achieved. In order to harness correctly this potential, it is clear that the physiological analysis of each strain studied needs to be undertaken under conditions as close as possible to the physico-chemical environment in which the strain evolves within the full-scale process. Furthermore, this analysis needs to be undertaken throughoutthe entire fermentation so as to take into account the changing environment in an essentially dynamic situation in which metabolic stress is accentuated by the microbial activity itself, leading to increasingly important stress response at a metabolic level. All too often these industrial fermentation constraints are overlooked, leading to identification of targets whose validity within the industrial context is at best limited. Thus the conceptual error is linked to experimental design rather than inadequate methodology. New tools are becoming available which open up new possibilities in metabolic engineering and the characterisation of complex metabolic networks. Traditionally metabolic analysis was targeted towards pre-identified genes and their corresponding enzymatic activities within pre-selected metabolic pathways. Those pathways not included at the onset were intrinsically removed from the network giving a fundamentally localised vision of pathway functionality. New tools from genome research extend this reductive approach so as to include the global characteristics of a given biological model which can now be seen as an integrated functional unit rather than a specific sub-group of biochemical reactions, thereby facilitating the resolution of complexnetworks whose exact composition cannot be estimated at the onset. This global overview of whole cell physiology enables new targets to be identified which would classically not have been suspected previously. Of course, as with all powerful analytical tools, post-genomic technology must be used carefully so as to avoid expensive errors. This is not always the case and the data obtained need to be examined carefully to avoid embarking on the study of artefacts due to poor understanding of cell biology. These basic developments and the underlying concepts will be illustrated with examples from the author's laboratory concerning the industrial production of commodity chemicals using a number of industrially important bacteria. The different levels of possibleinvestigation and the extent to which the data can be extrapolated will be highlighted together with the extent to which realistic yield targets can be attained. Genetic engineering strategies and the performance of the resulting strains will be examined within the context of the prevailing experimental conditions encountered in the industrial fermentor. Examples used will include the production of amino acids, vitamins and polysaccharides. In each case metabolic constraints can be identified and the extent to which performance can be enhanced predicted

• Journal of Acupuncture Research
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• 제23권3호
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• pp.177-190
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• 2006

Objectives : It has long been known about the anticancer effect of GRR-HAS, however, it has not been systemically determined the differentially regulated genes by GRR-HAS in cancer cells. The purpose of this study is to screen the GRR-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cell lines. Oligonucleotide microarray and proteomic approaches were employed to screen the differential expression genes. Methods : GRR~HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of GRR-HAS (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of GRR-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cells in all concentrations(0.1, 0.5, 1.5, 10,$20mg/m{\ell}$). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 320 with 6 up-regulated and 314 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down -regulated protein was protein disulfide isomerase and up-regulated proteins were fatty acid binding protein 1 and 14-3-3 gan1lTIa protein by $1.5mg/m{\ell}$ of CRR-HAS. Discussion : This study showed the comprehensive gene expression analysis using oligonucleotide microarray for the screening of GRR-HAS mediated differentially regulated genes. These results will provide a better application of GRR-HAS in cancer field and drug target development.

• Nutrition Research and Practice
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• 제9권1호
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• pp.79-86
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• 2015

BACKGROUND/OBJECTIVES: It is well-known that alcohol consumption is associated with stroke risk as well as with aldehyde dehydrogenase 2 gene (ALDH2) polymorphisms. However, it is unclear whether ALDH2 polymorphisms are associated with stroke risk independent of alcohol consumption and whether such association is modified by sex. We evaluated sex-specific associations of a common ALDH2 polymorphism and alcohol consumption with stroke risk in a Korean population. SUBJECTS/METHODS: We conducted a prospective cohort study involving 8,465 men and women, aged 40-69 years and free of stroke between June, 2001 and January, 2003, and followed for the development of stroke. We identified new cases of stroke, which were self-reported or ascertained from vital registration data. Based on genome-wide association data, we selected a single-nucleotide polymorphism (rs2074356), which shows high linkage disequilibrium with the functional polymorphism of ALDH2. We conducted Cox proportional hazards regression analysis considering potential risk factors collected from a baseline questionnaire. RESULTS: Over the median follow-up of 8 years, 121 cases of stroke were identified. Carrying the wild-type allele of the ALDH2 polymorphism increased stroke risk among men. The multivariate hazard ratio [95% confidence interval] of stroke was 2.02 [1.03-3.99] for the wild-type allele compared with the mutant alleles, but the association was attenuated after controlling for alcohol consumption. Combinations of the wild-type allele and other risk factors of stroke, such as old age, diabetes mellitus, and habitual snoring, synergistically increased the risk among men. Among women, however, the ALDH2 polymorphism was not associated with stroke risk. CONCLUSIONS: The prospective cohort study showed a significant association between a common ALDH2 polymorphism and stroke risk in Korean men, but not in Korean women, and also demonstrated that men with genetic disadvantages gain more risk when having risk factors of stroke. Thus, these men may need to make more concerted efforts to control modifiable risk factors of stroke.

• BMB Reports
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• 제39권1호
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• pp.68-75
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• 2006

In higher plants, P450s participate in the biosynthesis of many important secondary metabolites. Here we reported for the first time the isolation of a new cytochrome P450 cDNA that expressed in a stem-specific manner from Camptotheca acuminata (designated as CaSS), a native medicinal plant species in China, using RACE-PCR. The full-length cDNA of CaSS was 1735 bp long containing a 1530 bp open reading frame (ORF) encoding a polypeptide of 509 amino acids. Bioinformatic analysis revealed that CASS contained a heme-binding domain PFGXGRRXCX and showed homology to other plant cytochrome P450 monooxygenases and hydroxylases. Southern blotting analysis revealed that there was only one copy of the CaSS present in the genome of Camptotheca acuminata. Northern blotting analysis revealed that CaSS expressed, in a tissue-specific manner, highly in stem and lowly in root, leaf and flower. Our study suggests that CaSS is likely to be involved in the phenylpropanoid pathway.

• The Plant Pathology Journal
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• 제25권4호
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• pp.344-351
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• 2009

A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

• 한국식품과학회지
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• 제46권1호
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• pp.44-50
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• 2014

고압처리가 발아벼 추출물의 성인병관련 예방효과에 미치는 영향을 살펴보기 위하여 발아기간 및 고압처리 시간에 따른 효소저해활성을 살펴보았다. 발아기간은 6일로 하였고, 30 MPa의 압력 하에서 24시간 및 48시간 동안 처리하였다. ${\alpha}$-Glucosidase 저해활성은 발아초기에 대조구에 비해 고압처리 시 높은 범위의 저해활성을 나타내었으며, ${\alpha}$-amylase 저해활성은 발아기간 및 고압처리시간이 증가함에 따라 유의적으로 증가하였다. ACE 및 lipase 저해활성은 ${\alpha}$-glucosidase 저해활성과 유사하였으며, 고압처리가 대조구보다 높은 저해활성을 나타내었으며, 48시간 고압처리한 2일차 발아벼에서 가장 높은 저해활성을 나타내었다. Xanthine oxidase 저해활성은 발아초기에는 대조구보다 고압처리시 높았지만 발아가 진행됨에 따라 감소하였다. 이상의 결과로 부터 발아와 고압처리를 병행한 벼는 무발아 및 발아벼에 비하여 활용가치가 높을 것으로 판단되며, 성인병 예방을 위한 기능성식품소재로써 이용이 가능할 것으로 판단된다.

• 대한환경공학회지
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• 제34권12호
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• pp.855-862
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• 2012

Free-living 선충으로 알려진 Caenorhabditis elegans는 주로 토양 공극수에서 서식하며, 토양 영양단계, 에너지 흐름, 그리고 분해자로서 중요한 역할을 하는 것으로 알려져 있다. 최근 들어, C. elegans는 나노독성연구에 널리 사용되고 있는 추세이다. 본 연구에서는 C. elegans를 이용한 나노독성과 그에 대한 기작에 관련된 선행연구를 조사하였으며, 총 20건의 연구를 확인하였다. 대부분의 연구는 K-medium, S-medium, 그리고 NGM (Nematode Growth Medium) plate를 노출매체로 이용하고 있으며, 나노물질에 노출된 C. elegans로부터 관찰된 영향으로는 노화억제, 광독성영향, 유전독성, 그리고 표피자극 등이 포함되었다. C. elegans를 이용한 독성기작 연구는 개체 생활사 영향 평가, 산화스트레스 영향 평가, 유전독성영향 평가, 나노물질의 생체 내 분포, 그리고 나노물질 특성에 의한 독성영향 평가로 분류할 수 있다. 본 연구에서는 다양한 세포활성, 잘 알려진 유전정보, 그리고 투명한 구조로 인한 나노물질 관찰의 용이성을 바탕으로, C. elegans를 이용한 나노독성연구의 장점을 확인하였다. C. elegans는 나노독성을 평가하기에 적합한 시험종으로 고려되고 있다.

• 한국정보통신학회:학술대회논문집
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• 한국정보통신학회 2018년도 춘계학술대회
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• pp.588-591
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• 2018

Baculovirus는 원래 알팔파 루퍼 (looper)로부터 분리되었으며 154 개의 오픈 리딩 프레임 (ORF)을 가진 134-kbp 게놈을 포함하고 있다. 주요 캡시드 단백질 VP39는 약간의 단백질과 함께 p6.9 단백질로 DNA를 감싸는 뉴클레오 캡시드($21nm{\times}260nm$)로 형성된다. 그것들은 막대 모양의 캡시드 안에 이중 가닥의 고리 모양의 슈퍼 코일 DNA 분자이다. 야생형 baculovirus는 용균 및 폐색 된 생명주기를 모두 나타내며 바이러스 복제의 3 단계에 걸쳐 독립적으로 발달한다. 재조합 baculovirus는 광범위한 포유류 세포 유형에서 벡터를 전달하고 재조합 단백질을 발현 할 수 있다. 특히, 이들 baculovirus 벡터에 우세한 선별 마커를 포함시킴으로써 많은 세포에서 다양한 재조합 유전자를 발현시킬 수 있다. 본 연구의 배큘로 바이러스 벡터는 cytomegalovirus (CMV) 프로모터, uroplakin II promoter, polyhedron promoter, 수포 구내염 바이러스 G (VSVG), 녹색 형광 단백질 (EGFP), 단백질 전달 도메인 (PTD) 유전자 등으로 재구성되었다. 이러한 재구성 된 벡터를 다양한 세포 및 세포주에 감염시켰다. 우리는 다른 재조합 벡터와 비교하여 이러한 재조합 벡터의 전이 및 발현을 조사하는 수행하였다. 본 연구에서, 우리는 이 재조합 벡터의 형질 감염 및 발현이 어떤 대조군 벡터보다 더 높은 효능을 갖는다는 것을 알았다. 본 연구는 과학 기술부, 한국 정보 기술 진흥 기금 (MSIP)이 후원하는 한국 연구 재단 (NRF)을 통해 중견 연구원 프로그램 (NRF-2016R1A2B4016552)을 통해 지원되었다.

• 한국결정학회지
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• 제15권1호
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• pp.9-17
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• 2004

Trigonal system에 屬한 25個 space group들 中 18個는 hexagonal axes만을 나타내는 lattice letter p로 表示되고, 나머지 7個의 rhombohedral space groups R3(146), $R\={3}$(148), R32(155), R3m(160), R3c(161), $R\={3}m$(166), $R\={3}c$(167)은 첫째 3개의 lattice points (0, 0, 0), (2/3, 1/3, 1/3), (1/3, 2/3, 2/3)를 갖는 hexagonal cell in obverse setting과 둘째 primitive rhombohedral cell의 두 가지로 表示된다. 本 論文에서는 space group $R\={3}c$(167)에 대하여 論한 後 이 space group에 屬한 化合物인 tris(1,2,3,4-tetraphenylbuta-1,3-dienyl)cyclotriphosphazene, $C_{84}H_{60}N_3P_3$의 構造를 hexagonal 및 rhombohedral cell 의 양쪽으로 糾明하여 發表하였다.

• Genomics & Informatics
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• 제15권3호
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• pp.87-97
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• 2017

Multiple myeloma (MM) is a malignant disease caused by an abnormal proliferation of plasma cells, of which the prognostic factors include chromosomal abnormality, ${\beta}$-2 microglobulin, and albumin. Recently, the term chromothripsis has emerged, which is the massive but highly localized chromosomal rearrangement in response to a one-step catastrophic event. Many studies have shown an association of chromothripsis with the prognosis in several cancers; however, few studies have investigated it in MM. Here, we studied the association between chromothripsis-like patterns and treatment resistance or prognosis. First, we analyzed nine MM cell lines (U266, MM.1S, RPMI8226, KMS-11, KMS-12-BM, KMS-12-PE, KMS-28-BM, KMS-28-PE, and NCI-H929) and bone marrow samples of four patients who were diagnosed with MM by next-generation sequencing-based copy number variation analysis. The frequency of the chromothripsis-like pattern was observed in seven cell lines. We analyzed the treatment-induced chromothripsis-like patterns in KMS-12-BM and KMS-12-PE cells. As a result, breakpoints and chromothripsis-like patterns were increased after drug treatment in the relatively resistant KMS-12-BM. We further analyzed the patients' results according to the therapeutic response, which was divided into sensitive and resistant, as suggested by the International Myeloma Working Group. The chromothripsis-like pattern was more frequently observed in the resistant group. In the sensitive group, the frequency of the chromothripsis-like pattern decreased after treatment, whereas the resistant group showed increased chromothripsis-like patterns after the treatment. These results suggest that the chromothripsis-like pattern is associated with treatment response in MM.