• Microbiology and Biotechnology Letters
• /
• v.29 no.2
• /
• pp.72-77
• /
• 2001

Expression of f3 ~agarase Gene and Catabolite Repression in Escherichia coli by the Promoter of Alginate Lyase Gene Isolated from Marine Pseudomonas sp. Jin, Cheal~Ho, J~Hyeon Park, Jeong-Hyun Han, YoonM Hyeok Chae, Jong~Hee Lee, Jung-Kee Lee!, and In-800 Kong*. Faculty of Food Science and Biotechnology, Pukyong National UniversitYt Pusan 608-737, Korea, llnBioNet Co. 1690-3 Taejon 306-230, Korea - Promoter is a key factor for expression of the recombinant protein. There are many promoters for overexpression of protein in various organisms. The aly promoter of Pseudomonas sp. W7 isolated from marine environment was known to be a constitutive expression promoter of the alginate lyase gene, and it's promoter activity is repressed by glucose in Escherichia coli. To investigate the catabolite repression of the aly promoter ~md association between the promoter mutants, f3 agarase gene, which was also cloned from Pseudomonas sp. W7 was connected to the aly promoter with the sequence the coding 46 N-terminal amino acids ofthe alginate lyase gene. The constructed plasmid was introduced into E. coli and the agarase activity was measured. Fourty six amino acids of the alginate lyase gene was serially deleted using peR to the direction of 5' upstream region and subcloned. The agarase was overexpressed by the aly promoter and the production of agarase was repressed by the addition of glucose into culture media. Fourty six amino acids of alginate lyase did not affect the production of agarase at all. The deletion of a putative stem-loop structure in the aly promoter induced the decrease of f3 -agarase productivity.

• Proceedings of the Korean Information Science Society Conference
• /
• 2004.04b
• /
• pp.253-255
• /
• 2004

다량의 유전자 발현 정보를 담고 있는 DNA 마이크로어레이 기술의 발달로 인해 대량의 생물정보를 한번의 실험을 통해 분석할 수 있게 되었다. 유전자 발현 데이터를 분석하는 방법 중 하나인 클러스터링은 비슷한 기능을 가진 유전자들을 그룹별로 묶어서 그룹 레의 유전자들의 기능을 밝히거나 미지의 유전자를 분석하는데 이용되고 있다 본 논문에서는 유전자 발현 데이터를 클러스터링 하여 그로부터 유전 정보를 찾아내기 위한 방법으로 GG (Gath-Geva) 알고리즘을 제시한다. 퍼지 클러스터링 알고리즘중 하나인 GG 알고리즘은 대표적인 퍼지 클러스터링 방법인 퍼지 c-means 와 GK (Gustafson-Kessel) 알고리즘을 개선한 것으로. 차원이 크고 분포가 애매하여 클러스터링이 어려운 유전자 발현 데이터의 클러스터링에 적합한 알고리즘이다. 혈청(Serum) 유전자 데이터와 효모(Yeast) 세포주기 데이터를 CG 알고리즘 이용해 클러스터링 해 보고, 그 결과를 퍼지 c-means 알고리즘, GK알고리즘과 비교해 본 결과, GG 알고리즘이 유전자 발현 데이터의 클러스터링에 더 적합함을 확인하였다.

• Journal of the Korean Recycled Construction Resources Institute
• /
• v.2 no.3
• /
• pp.239-246
• /
• 2014

High volume slag concrete is attracting new attention and are thought to have promising potential for industrial applications, partly due to the climate debate, but especially due to their very low heat of hydration and their good durability in chemically aggressive environments. However, High volume slag concretes tend to have slower strength development especially. In this study, the effect of anhydrite ($CaSO_4$) on the mechanical and durability performance of high volume slag concrete were investigated. The main variables were anhydrite contents (0, 4, 6, 8, 10%). Test results show that 4~8% anhydrite concrete have improved engineering properties (hydration, compressive strength, shrinkage, creep, carbonation) as control concrete at early ages.

• Microbiology and Biotechnology Letters
• /
• v.26 no.1
• /
• pp.15-22
• /
• 1998

The dnrF gene, responsible for conversion of aklavinone to $\varepsilon$-rhodomycinone via C-11 hydroxylation, was mapped in the daunorubicin gene cluster of Streptomyces peucetius subsp. caesius ATCC 27952, close to drrAB, one of the anthracycline resistance genes. To characterize the enzymatic properties of the aklavinone 11-hydroxylase, the dnrF gene was overexpressed in Escherchia coli. The pET-22(+) plasmid which has the T7 promoter under the control of lacUV5 gene was used for the overexpression of the dnrF gene, and the recombinant plasmid pET213 that contains the dnrF gene linked to the T7 promoter of pET-22b(+) was introduced into the E. coli BL2l. When the expression of the dnrF gene was induced by IPTG at the final concentration of 1 mM, the induced protein could be detected in SDS-PAGE only in insoluble precipitate. The insoluble protein was electroeluted from the gel and used for the preparation of antiserum in mice. Various culture conditions were tested to maximize the expression of the aklavinone 11-hydroxylase in soluble form. The enzymatic activity was checked by the bioconversion experiment, and the protein was confirmed by the SDS-PAGE and the Western blot analysis. From the analysis of the data, it was concluded that the culture induced with IPTG at the final concentration of 0.02 mM at 37$^{\circ}C$ yielded the best productivity of active form of enzyme.

• Proceedings of the Korean Information Science Society Conference
• /
• 2004.04b
• /
• pp.244-246
• /
• 2004

마이크로어레이 칩 실험을 통하여 대량으로 생산되는 유전자 발현 데이터는 여러 가지 클러스터링 방법을 적용하여 분석할 수 있으며, 생성된 클러스터들 또한 여러 가지 방법으로 해석 할 수 있다. 본 논문에서는 기존의 클러스터링 방법들을 응용한 seed클러스터링 방법을 제안하고 생물학적 온톨로지인 Gene Ontology를 기반으로 클러스터를 해석한다. 본 논문에서는 효과적인 유전자 발현 데이터 클러스터링 방법과 생물학적 지식을 바탕으로 클러스터를 해석, 평가하는 방법을 보여 준다.

• The Korean Journal of Zoology
• /
• v.33 no.3
• /
• pp.303-309
• /
• 1990

In order to have a handle on the availability of eukarvotic peptide hormone precursors, a cDNA encoding angler fish prepro-SRIF I was manipulated so that it can be produced in large quantity from heterologous E. coli cells. Using T7 overexpression system, fusion constructs between the T7 phage coat protein Sl0 and the prepro-SRIF were made and modified as desired. From the host E. coli strain, BL21 DE3, harboring these plasmid constructs, three different SRIF related polypeptides were expressed in large amount and characterized. The results confirm the exact construction and authenticity of the overexpressed proteins from E. coli cells. The importance of this heterologous overexpression in hard to get peptide hormone precursors as well as the suitability of the target peptide hormone SRIF for this approach are discussed.

• Proceedings of the Korean Information Science Society Conference
• /
• 2005.11b
• /
• pp.799-801
• /
• 2005

현재 마이크로어레이 기술은 대량의 유전자 발현 데이터 특히 종양과 관련한 데이터들을 쏟아내고 있다. 이 데이터를 기반으로 종양의 종류에 따른 유전자들의 차별적 발현 양상을 분석하고 발현량의 변화가 두드러지는 유전자들에 기반하여 종양을 분별할 수 있는 분류 모델을 구축한 후, 이것을 종양을 진단하거나 예측하는데 이용할 수 있다. 대부분의 종양은 생성 매커니즘에 따라 세부 부류로 나눌 수 있고 세부 부류에 따라 치료 방법이나 예후가 달라지므로, 정확하게 종양의 세부 부류를 진단하는 것이 매우 중요하다. 본 논문에서는 종양의 종류에 따라 발현량이 민감하게 변화하는 유전자들을 뽑아내기 위한 특징 추출 방법들과 추출된 특징들에 기반해서 종양의 종류를 분별할 수 있는 기계학습 알고리즘들의 조합들의 성능을 비교분석 하였다.

• Journal of Life Science
• /
• v.14 no.3
• /
• pp.435-440
• /
• 2004

The gene encoding a extremely thermostable 4-$\alpha$-glucanotransferase from a hyperthermophilic archaeon, Thermococcus litoralis, was cloned, sequenced and expressed in Escherichia coli. The amino acid sequence of the enzyme was distantly related to other functionally-related ones, such as D-enzymes. The enzyme is of industrial interest because of a novel activity of producing cycloamylose and is also important for fundamental studies of protein, sugar-metabolizing enzymes. In this paper, the overexpression of 4-$\alpha$-glucanotransferase in E. coli was carried out expression vector system with lac and T7 promoters. The enzyme was successfully overexpressed, and purified by the heat treatment of a cell-free extract, successive Butyl-Toyopearl and Mono Q chromatographies. The purified recombinant enzyme showed the same specific activity and the same mobility in SDS-PAGE as natural enzyme.

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.711-714
• /
• 2001

The production of the recombinant Plasmodium vivax merozoite surface protein (PvMSP) has been investigated in the recombinant E.coli system. Experimental optimization of the culture conditions, such as the effect of initial pH, and operating temperature has been tried on the growth of recombinant E.coli and on the overproduction of the target foreign protein.

• Proceedings of the Korean Information Science Society Conference
• /
• 2006.06b
• /
• pp.220-222
• /
• 2006

현재 마이크로어레이 기술은 대량의 유전자 발현 데이터 특히 암과 관련한 데이터들을 쏟아내고 있다. 이 데이터를 기반으로 암의 종류에 따른 유전자들의 차별적 발현 양상을 분석하고 발현량의 변화가 두드러지는 유전자들에 기반하여 암을 분별할 수 있는 분류 모델을 구축한 후, 이것을 암을 진단하거나 예측하는데 이용할 수 있다. 본 논문에서는 마이크로어레이 데이터를 사용해 특징추출방법과 분류를 위한 Naive Bayes, k-Nearest Neighborhood, Decision Tree, Support Vector Machine, Neural Network 알고리즘을 이용하여 최적의 조합을 찾고 어떤 알고리즘이 가장 효과적인지 실험을 통해 분석해보고 성능평가 하는 것을 목표로 한다.