• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.231-231
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• 2005

• 한국작물학회지
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• 제49권spc1호
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• pp.307-317
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• 2004

• 한국작물학회:학술대회논문집
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• 한국작물학회 2003년도 춘계 학술대회지
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• pp.51-51
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• 2003

• Journal of Microbiology and Biotechnology
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• 제6권5호
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• pp.361-363
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• 1996

The terminal regions of the double-stranded RNA (dsRNA) genome segments of infectious pancreatic necrosis virus (IPNV) DRT strain were sequenced. The dsRNAs, which were $^{32}P$-labelled at their 3'-termini by incubation with [$^{32}P$]pCp and T4 RNA ligase, were separated by 5$%$ polyacrylamide gel electrophoresis, and the segments A and B of IPNV-DRT were sequenced by two-dimensional gel electrophoresis. The 5'-terminal sequences of the IPNV-DRT plus strand from two genome segments were found to have the same conserved nucleotide (5'-CGG(C/A)A-), but the 3'-terminal sequences -CCCCAGGCG-3' and -CGGACCCCG-3' were found in the plus strand from segments A and B, respectively. The inverted oligonucleotide sequences of 3'-terminal of between segments A and B were found and they differ from those of other IPNVs.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1222-1228
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• 2006

Two kinds of Haematococcus pluvialis cells (green vegetative cells cultivated under optimal cell culture conditions and red cyst cells maintained under high light stress conditions to induce astaxanthin production) were used to investigate the protein expression profiles by two-dimensional electrophoresis, image analysis, and peptide mass fingerprinting. The cellular accumulation of astaxanthin was evident after exposure to high light intensity and reached the maximum cellular level after 78 h of high light stress. In a 2-D electrophoresis analysis, 22 proteins were upregulated over 2-fold in the red cyst cells when compared with the green vegetative cells and selected for further analysis by chemically assisted fragmentation (CAF)-MALDI-TOF sequencing to identify the protein functions. Among 22 different spots, several key enzymes specific to the carotenoid pathway, including isopentenyl pyrophosphate isomerase (IPP) and lycopene $\beta$-cyclase, appeared in H. pluvialis after exposure to high light intensity. Therefore, IPP and lycopene $\beta$-cyclase would appear to be involved with carotenoid accumulation in the cytoplasm, as these peptides were preferentially upregulated by high light intensity preceding an increase in carotenoid, and only these forms were detected in the red cyst cells.

• 정보처리학회논문지B
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• 제12B권3호
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• pp.239-246
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• 2005

생물학자가 단백질을 검색하고 분석하기 위해서는 2차원 젤 전기영동(2DGE : Two Dimensional Gel Electrophoresis) 실험을 해야 한다. 실험 결과는 2차원 영상이 생성된다. 2차원 영상에서 단백질 반점의 패턴 분석을 위해 2차원 젤 영상에 펼쳐진 단백질 반점들을 영상처리를 통해 분할하고, 대조 그룹의 단백질 패턴과 비교분석을 통해 밝히고자하는 단백질 반점을 찾아내야 한다. 단백질 반점을 분할하는 알고리즘에 있어서 기존에는 가우시안 함수를 적용하였지만, 최근 들어 형태학 분리개념에 의한 Watersheds 영역기반 분할(Watersheds region-based segmentation) 알고리즘을 활용하고 있다. 그러나 Watersheds 영역기반 분할 알고리즘은 크기가 큰 영상에서 원하는 영역을 신속하게 분할한다는 장점이 있지만, 영상 화소의 그레이 값이 연속적인 경우 실제 반점의 개수 에 비해 과다분할(over-segmentation)되거나 과소분할(under-segmentation)의 문제점을 안고 있다. 이는 마커(marker) 포인트의 설정에 의해 어느 정도 해결할 수 있지만 병합(merge)과 분할(split) 과정을 반복해야 한다. 본 논문은 Watersheds 기반 계층적 이진화 기법을 적용하여 마커 드리븐 Watersheds 영상분할의 문제점을 해결하고자 한다.

• 정보처리학회논문지D
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• 제12D권5호
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• pp.719-728
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• 2005

2DE는 조직 내의 단백질을 규명하는 단백질 분리 기술이다. 그러나 2DE 이미지는 실험 조건, 스캐닝 상태와 같은 환경에 민감하게 영향을 받는다. 이러한 이미지간의 변화를 극복하기 위해서 사용자는 각각의 서로 다른 이미지에 수동으로 기준점을 입력해주어야 한다. 그러나 이 과정은 에러를 발생시키며 긴 시간을 요구하는 작업으로, 빠른 분석에 장애 요인이 된다. 따라서 본 논문에서는 기준점 프로파일에 기반 하여 기준점을 자동으로 추출하는 방법을 개발하였다. 기준점 프로파일은 이미 확인된 이미지들의 기준점들에 대한 클러스터링 방법을 통하여 생성하며, 각 클러스터의 다양한 속성을 정의한다. 새로운 이미지가 입력되면 기준점의 후보 스팟들을 대상으로 프로파일과 비교하석 기준점을 추출한다. 그리고 $A^*$알고리즘을 이용하여 기준점 선정 과정을 최적화한다. 본 논문에서는 실제 사람의 간 조직 이미지를 이용하여 기준점 추출 방법의 성능을 분석하였다

• Journal of mucopolysaccharidosis and rare diseases
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• 제3권1호
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• pp.20-27
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• 2017

Purpose: Mucopolysaccharidosis IV (MPS IV) is a disease characterized by deficient activity of N-acetylgalactosamine-6-sulfatase (GALNS) causing excessive lysosomal storage of keratan sulfate (KS). The identification of the relevant disaccharide units of KS after keratanase II digestion followed by liquid chromatography/tandem mass spectrometry detection (LC-MS/MS) is validated and applicable for the preliminary diagnosis of MPS IV. Methods: A total of 67 urine samples were collected and analyzed from 11 MPS IV patients comprising 10 MPS IVA and one MPS IVB patients, and 56 normal controls. Urinary glycosaminoglycan was first precipitated by the Alcian blue method followed by a digestion of keratanase II. The protonated species of the digested disaccharide products were detected by using multiple reaction monitoring experiment. Results: One particular disaccharide of KS was selected. The transition mass-to-charge (m/z) of the parent ion and its daughter ion after collision was $462.0{\rightarrow}97.0$, whereas the chondrosine used as an internal standard in this assay was m/z $353.9{\rightarrow}73.0$. The results corresponded well with the two-dimensional electrophoresis method. The quantities of urinary KS were significantly raised in confirmed MPS IV patients when comparing with those of normal controls ($170.2{\pm}81.1$ vs. $4.06{\pm}1.92{\mu}g/mL$). Conclusion: The LC-MS/MS method for MPS IVA determination is specific, sensitive, validated, and applicable for urinary KS quantification. This method can be used not only as a first-line biochemistry examination of MPS IVA, but also as an outcome survey after enzyme replacement therapy.

• BMB Reports
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• 제39권2호
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• pp.171-177
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• 2006

Nonsteroidal anti-inflammatory drugs such as indomethacin (IN) can exert anti-colorectal cancer (CRC) activity through cyclooxygenase independent mechanism, but the exactly biological mechanism is not completely known. Here we use proteomic tools to investigate the molecular mechanism of this action. First, nude mice bearing tumors derived from subcutaneous injection with human CRC cell line HCT116 were randomly allocated to groups treated with or without indomethacin. Later, tumor lumps were incised and then total proteins extracted. After separated with two-dimensional electrophoresis, thirty-one differently expressed spots were found between IN-treated and non-IN-treated groups, of which 25 spots decreased and 6 spots increased in abundance in IN-treated group. Through matrix-assisted laser desorption ionization time of flight mass spectrometry and then NCBInr and SWISS-PROT databases searching, 12 protein spots were finally identified including galectin-1, annexin A1, annexin IV, trancription factor BTF3A, calreticulin. Most of the identified proteins are correlated with tumor's biological prosperities of proliferation, invasion, apoptosis and immunity, or take part in cell's signal transduction. From above we thought that indomethacin can exert its effect on colorectal cancer through regulating several proteins' expression directly or indirectly. Further study of these proteins may be helpful in founding new targets of drugs for cancer chemotherapy.

• Journal of Microbiology and Biotechnology
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• 제16권6호
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• pp.911-920
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• 2006

[ $CD8^+$ ] T Iymphocytes with the cytotoxic activity and capability to release various cytokines are the major players in immune responses against viral infection and cancer. To identify the proteins specific to resting or activated human CD8$^+$ T cells, human CD8$^+$ T cells were activated with anti-CD3+anti-CD28 mAb in the presence of IL-2. The solubilized proteins from resting and activated human CD8$^+$ T cells were separated by high-resolution two-dimensional polyacrylamide gel electrophoresis, and their proteomes were analyzed. Proteomic analysis of resting and activated T cells resulted in identification of 35 proteins with the altered expression. Mass spectrometry coupled with Profound and SWISS-PROT database analysis revealed that these identified proteins are to be functionally associated with cell proliferation, metabolic pathways, antigen presentation, and intracellular signal transduction pathways. We also identified six unknown proteins predicted from genomic DNA sequences specific to resting or activated CD8$^+$ T cells. Protein network studies and functional characterization of these novel proteins may provide new insight into the signaling transduction pathway of CD8$^+$ T cell activation.