• Proceedings of the KSR Conference
• /
• 2011.10a
• /
• pp.2672-2678
• /
• 2011

Carbody of rolling stock has been gradually changed as whole wood, steel frame with wood car body, whole steel car body with rivet and whole monocoque carbody with welding. And also mild steel has been used widely to material of structure, but usage of stainless and aluminium which have lightweight and high corrosion resistance is being increased lately. Structure is being commercialized to AED(All Extrusion Design),whole double skin with hollow excluded shape such as aluminium structure from SSD(Sheet Stringer Design), single skin consists of traditional frame and outside plat. Traditional aluminium carbody had many problems from reduced strength in welding combination section because car body is consist of small extruded material affected heat by welding. On this study, we proposed the plan to improve the body strength and quality with large-scale aluminium extruded material by minimizing welding concentration in combination section.

• Journal of Life Science
• /
• v.7 no.3
• /
• pp.161-166
• /
• 1997

HIV-1 Rev protein plays an important role in regulating the expression of viral structural proteins. It allows the nuclear export and accumulation of unspliced and partially spliced viral mRNA in the cytoplasm. The Rev-responsive element RNA, present in the env gene, forms a higly ordered RNA secondary structure and is required for the Rev-mediated mRNA export. For this process to complete factor(s) are strongly suggested. From our experiments of electrophoretic mobility shift, UV-crosslinking and SDS/PAGE, RRE RNA was found to be recognized to several nuclear factors such as 36/37, 56, 41. 76, 150 kD proteins in the order of reactivity. Among them, 36/37 and 56 kD proteins are more reactive upon a brief UV treatment (5 min) and more persistent in the presence of high amount of nonspecific competitor, heparin. Certain nuclear protein9s) seemed to recognize the RRE RNA structure in competition with Rev to gel mobility shift assay.

• Journal of Life Science
• /
• v.28 no.3
• /
• pp.275-283
• /
• 2018

Human fibroblast growth factor (FGF) has the potential to be a commercially important therapeutic or cosmeceutical agent due to its ability to generate tissue and heal wounds. Granting permeability into skin tissues increases the therapeutic effects of FGF. Thus, several researchers have attempted the fusion of FGF conjugates with protein transduction domains (PTDs) to investigate the transduction ability and therapeutic effects of FGF. Less is known, however, about whether the location of PTD fused to the N- or C-terminus of FGF proteins has a significant impact on the folding and stability in Escherichia coli, and eventually, on transduction. Here, we report cloning of human basic fibroblast growth factor (FGF2) as a control and FGF2 with PTD fused to the N- or C-terminal ends of FGF proteins by an overlap extension PCR. We performed expression, verified expression properties of recombinant FGF2 without or with PTD fused to the N-terminus and the C-terminus, and investigated transduction ability into tissue by treating the dorsal skin of mice subjects. As a result, FGF2 and FGF2-PTD (fused to C-terminus) fusion protein were expressed as soluble forms suitable for straight-forward purification, unlike insoluble PTD-FGF2 (fused to N-terminus), but only FGF2-PTD fusion protein could transduce into the dorsal skin tissue of the mice subjects. Our results suggest that FGF2 with PTD fused to the C-terminus is more efficient than other options in terms of expression, purification, and delivery into skin tissue, as it does not require labor-intensive, costly, and time-consuming methods.

• Microbiology and Biotechnology Letters
• /
• v.33 no.2
• /
• pp.96-105
• /
• 2005

For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

• Proceedings of the Optical Society of Korea Conference
• /
• 2003.02a
• /
• pp.334-335
• /
• 2003

현재 기지국과 중계기 사이의 광선로에 사용되고 있는 1310 nm와 1550 nm 파장이외에 OADM (optical add-drop multiplexer)을 사용하여 새로운 2개 파장을 광선로에 추가로 연결함으로써 광선로 임차사용의 효율을 증대시키는 4파장 광중계기의 구현시 drop신호는 삽입손실 ­3 dB 이하의 저손실특성이 요구되며 isolation 신호는 통상 ­40 dB 이상의 고품질이 요구된다. 그러나 이와같은 성능의 광모듈을 실제로 구현하기 어렵다. 따라서 많은 광결합 부품의 조합을 통해서도 drop또는 isolation 특성은 4파장 광중계기를 실용화하기 어려울 만큼 저하된다. (중략)

• Proceedings of the Korea Information Processing Society Conference
• /
• 2012.04a
• /
• pp.1141-1144
• /
• 2012

최근의 SW 개발환경은 네트워크 환경이 발달하고 복잡한 비즈니스를 구현하기 위해 분산 컴퓨팅 환경으로 옮겨진 상황이다. 따라서, 시스템간의 쉬운 연결성을 보장하여 자원의 낭비를 줄이고 재사용하며 활성화 할 수 있는 플랫폼을 갖출 수 있는 SOA(Service Oriendted Architecture)의 활용이 필요한 시점이다. 이런 복잡한 니즈의 비즈니스를 구현함에 있어 업무 규칙을 별도로 관리하고 룰엔진에 의해 판단을 할 수 있다면, 프로그램 코드로부터 업무를 완전히 분리해낼 수 있다 본 논문에서는 이러한 진일보한 사상을 조합하여 비즈니스 구현에 집중하고 결합성은 낮출 수 있는 SW 개발 아키텍쳐를 제시해본다.

• Proceedings of the Korea Information Processing Society Conference
• /
• 2010.11a
• /
• pp.391-394
• /
• 2010

기존의 M&S 프레임워크는 컴포넌트 간에 송수신되는 메시지의 구조가 유연하지 못한 문제점을 가지고 있다. 플러그인 기반 아키텍처는 소프트웨어를 구성하는 컴포넌트들을 플러그인 형태로 구현하여 컴포넌트간의 결합도를 낮추고 유연성 및 재사용성을 강화할 수 있는 구조를 가지고 있다. 이러한 아키텍처를 구성하는 각 컴포넌트는 메시지 지향 미들웨어 기반의 메시지 통신을 수행하게 되는데, 플러그인 간에 종속성이 생기지 않는 형태로 설계되고 구현되어야 한다. XML기반의 객체모델은 이러한 메시지 통신에 사용되는 메시지 객체의 구조를 정의한다. 이 객체모델은 사용자 정의 형식을 지원하며 이러한 형식을 조합하여 새로운 복합 형식을 정의하여 메시지 구조를 표현할 수 있도록 한다. 객체 모델에서는 각 사용자 정의 형식과 각 형식에서 사용하는 기본형(Primitive Type)의 클래스를 추상화하여 정의함으로써 객체 모델의 유연성을 높일 수 있는 구조를 가지고 있다.

• Korean Journal of Microbiology
• /
• v.30 no.3
• /
• pp.218-224
• /
• 1992

Recombinant plasmid shuttle vectors were constructed for genetic studies on the oxidation of carbon monoxide by carboxydobacteria. Two vectors. pYK322 (7.2 kb, Ap'. Tc') and pYK 324 (7.2 kb, Ap', Tc'), were constructed using pBR322 and pYK100. a small plasmid in Pseudomonas carbo,xydovorans. Four plasmids. pYK2IO (5.2 kb. Cm'), pYK220 (5.2 kb, Cmr), pYK230 (5.2 kb, Cm'), and pYK232 (5.2 kb. Cm'), were constructed using pACYC184 and pYK100. Transformation of several carboxydobacteria with pYK322 and pYK220 was round to be efficient when the cells were transformed by the methoti of Bagdasarian and Timmis (Curr. Top. Microbiol. Immunol. 96:47-67. 1982) with several modifications; cells growing on 0.2% succinate were harvested at the mid-exponential phase. 10 mM RbCl in transformation solution was substituted with 100 mM KCI. cclls in transformation solution were incubated for 12 h at 4'C before addition of DNA and heat shock was carried out for 3 min at 45$^{\circ}$C. Plasmid vectors used for transformation, however. were not detected from antibiotics-resistant transformants, suggesting that the vectors may be integrated into the chromosomal DNA.

• Microbiology and Biotechnology Letters
• /
• v.40 no.2
• /
• pp.92-97
• /
• 2012

Antimicrobial peptides (AMPs) are important components of living organisms acting against Gram-negative and Gram-positive bacterial and fungal pathogens. Cathelicidin human peptides have a variety of biological activities that can be used in clinical applications. AMPs are not produced naturally in large quantities, and chemical synthesis is also economically impractical, especially for long peptides. Therefore, as an alternative, heterologous expression of AMPs by recombinant techniques has been studied as a means to reduce production costs. E. coli is an excellent host for the expression of AMPs, as well as other recombinant proteins, because of the low cost involved and its easy manipulation. However, overexpression of AMPs in E. coli has been shown to cause difficulties resulting from the toxicity of the subsequently produced AMPs. Therefore, fusion expression was theorized to be a solution to this problem. In this study, AMPs were expressed as fused proteins with the glutathione S-transferase (GST) binding protein to protect against the toxicity of AMPs when expressed in E. coli. The LL37, and hybrid gaegurin and LL37 (GGN4(1-16)-LL37(17-32), which we designated as GL32, peptides were expressed as GST-fusion proteins in E. coli and the fusion proteins were then purified by affinity columns. The purified peptides were obtained by removal of GST and were confirmed by western blot analysis. The purified antimicrobial peptides then demonstrated antimicrobial activities against Gram-negative and Gram-positive bacterial strains.

• KSBB Journal
• /
• v.10 no.5
• /
• pp.475-481
• /
• 1995

The effect of metallothionein expression on the metal resistance and removal by recombinant Saccharomyces cerevisiae containing the plasmid pJW9 was investigated. The recombinant strain S. cerevisiae BZ-pJ was constructed by transforming the host strain S. cerevisiae BZ3l-1-7Ba with the gene coding for a metal-binding protein, metallothionein. Introduction of the MT gene yielded an increase in the minimum inhibitory concentration (MIC) of copper more than three times compared with the host strain. The minimum inhibitory concentrations of $Cr^{2+}, Znr^{2+} and Pb^{2+}, $ were not different for the two strains. The recombinant yeast grown in a medium containing 8mM CuSO4 was able to remove copper with a capacity of 18.9mg $Cu^{2+}$/g dry cell. In a mixture of copper and zinc, the presence of copper relieved the toxic effects caused by zinc, resulting in an enhancement of the final cell density and the specific growth rate of the recombinant yeast. The capability to remove copper by the recombinant yeast was linearly proportional to the copper concentrations in the medium. The efficiency of copper removal was rather constant regardless of the initial copper concentrations. The specific removal of zinc was dependent on the zinc concentrations in media, though, and such dependence was not so pronounced as the concentration of copper.