• Title/Summary/Keyword: sprC

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Distinct Regulation of the sprC Gene Encoding Streptomyces griseus Protease C from Other Chymotrypsin Genes in Streptomyces griseus IFO13350

  • Choi, Eun-Yong;Oh, Eun-A;Kim, Jong-Hee;Kang, Dae-Kyung;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.81-88
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    • 2007
  • The sprC gene encodes Streptomyces griseus protease C (SGPC), a bacterial chymotrypsin-like serine protease. Because the published data on sprC was not complete, we cloned and analyzed a new DNA fragment spanning downstream to upstream of the sprC gene from S. griseus IFO13350. The cloned 2.3-kb DNA fragment was placed on a high-copy number plasmid and introduced into Streptomyces lividans TK24. Chymotrypsin activity of the transformant was 8.5 times higher than that of the control after 3 days of cultivation and stably maintained until 9 days of cultivation, which dearly indicated that the cloned 2.3-kb fragment contained the entire sprC gene with its own promoter. When the same construct was introduced in the S. griseus IFO13350 (wild strain) and its two mutant strains in the A-factor regulatory cascade, ${\Delta}adpA$ and HO1, the chymotrypsin activity increased fivefold only in the ${\Delta}adpA$ strain. Transcriptional analysis based on RT-PCR revealed that the sprC gene is normally transcribed in both strains; however, earlier transcription was observed in the wild strain compared with the ${\Delta}adpA$ strain. A gel mobility shift assay showed that the AdpA protein did not bind to the promoter region of sprC. All these data clearly indicate that the expression of sprC is not dependent on the AdpA protein, but is distinctly regulated from other chymotrypsin genes composing an AdpA regulon. Earlier morphological differentiation was observed in S. lividans TK24, and S. griseus IFO13350 and HO1, transformed with the expression vector. The transformant of S. griseus ${\Delta}adpA$ formed markedly larger colonies. Antisense repression of sprC resulted in severe decrease of chymotrypsin activity, down to one-third of the control, and delayed morphological differentiation. All these data suggest that SGPC is related to normal morphogenesis in S. griseus.

Surface Plasmon Resonance Multisensing Using Thickness Difference of Additional Layer (부가층의 두께 차이를 이용한 표면플라즈몬공명 멀티센싱)

  • Kim, Young-Gyu;Oh, Myung-Hwan;Lee, Seung-Ki
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.55 no.10
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    • pp.492-498
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    • 2006
  • A novel surface plasmon resonance(SPR) multisensing method, which does not require imaging apparatus such as CCD, has been proposed and implemented experimentally. The proposed method is based on the multichannel SPR and the separation of signals by use of additional layers whose thickness is controlled. SPR signals are influenced by the thickness of sensing layer as well as the optical condition of sensing surface. As the SPR signals from different ligands are usually positioned closely, the reflected light from sensing surface does not provide us with the clear differences of resonance signal depending on the kinds of ligands. It was found from our experiments that SPR signals from each ligand that is located on the additional layer with different thickness can be separated clearly enough to identify various signals from different ligands. Proposed method with theoretical design and simulation has been verified experimentally by using $SiO_2$ thin film layer as additional layer.

Development of Protein Chip for Diagnosis of Chlamydophia Pneumoniae (단백질 칩을 이용한 클라미디아 폐렴의 진단)

  • Kim, Woo Jin;Lee, Hui Young;Lee, Seung-Joon;Jung, Se-Hui;Yuk, Jong Seol;Ha, Kwon-Soo;Jung, Ki-Suck
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.4
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    • pp.412-418
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    • 2006
  • Background; The diagnosis of chlamydial infection is based on serology. The current gold standard of diagnosis is MIF(microimmunofluorescence), but this modality is subjective and time-consuming. Protein microarray with using a SPR(surface plasmon resonance) sensor has recently been suggested as a method for detecting infection. For developing a protein chip to diagnose chlamydial infection, EBs(elementary bodies) were immobilized on a gold chip and the interaction between an antibody for Chlamydophila pneumoniae and the EBs(elementary bodies) immobilized on the surface of the gold chip was measured by using an SPR sensor. Methods; For the surface antigen, the EBs of Chlamydophila pneumoniae LKK1 were purified. Charged arrays were prepared by using PDDA(polydiallyldimethylammonium chloride) which has a positive charge. After immobilization of the chlamydial EBs on the PDDA surface, the investigation of the surface was done with using atomic force microscopy. After the antibody for C. pneumoniae was applied on chip, we monitored the SPR wavelength-shift to detect any antigen-antibody interaction with using a self-assembled SPR sensor. Results; The chlamydial EBs on the positively charged PDDA were visible on the surface with using atomic force microscopy. The SPR wavelength increased after interaction of antibody for C. pneumoniae with the EBs immobilized on charged gold surface. The wavelength-shift was correlated with the concentration of antigens. Conclusion; The surface immobilization of EBs on the gold surface with the charged arrays was identified and the antigen-antibody interaction on the gold chip was detected via the SPR sensor. Further investigations are needed to apply this technique to the clinical field.

Graphene Coated Optical Fiber SPR Biosensor

  • Kim, Jang Ah;Hwang, Taehyun;Dugasani, Sreekantha Reddy;Kulkarni, Atul;Park, Sung Ha;Kim, Taesung
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.401-401
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    • 2014
  • In this study, graphene, the most attractive material today, has been applied to the wavelength-modulated surface plasmon resonance (SPR) sensor. The optical fiber sensor technology is the most fascinating topic because of its several benefits. In addition to this, the SPR phenomenon enables the detection of biomaterials to be label-free, highly sensitive, and accurate. Therefore, the optical fiber SPR sensor has powerful advantages to detect biomaterials. Meanwhile, Graphene shows superior mechanical, electrical, and optical characteristics, so that it has tremendous potential to be applied to any applications. Especially, grapheme has tighter confinement plasmon and relatively long propagation distances, so that it can enhance the light-matter interactions (F. H. L. Koppens, et al., Nano Lett., 2011). Accordingly, we coated graphene on the optical fiber probe which we fabricated to compose the wavelength-modulated SPR sensor (Figure 1.). The graphene film was synthesized via thermal chemical vapor deposition (CVD) process. Synthesized graphene was transferred on the core exposed region of fiber optic by lift-off method. Detected analytes were biotinylated double cross-over DNA structure (DXB) and Streptavidin (SA) as the ligand-receptor binding model. The preliminary results showed the SPR signal shifts for the DXB and SA binding rather than the concentration change.

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A portable surface plasmon resonance sensor system for detection of C-reactive protein using SAM with dimer structure (소형 표면 플라즈몬 공명 센서와 이합체 구조를 가진 SAM을 이용한 CRP 검출)

  • Sin, Eun-Jung;Joung, Eun-Jung;Jo, Jin-Hee;Hwang, Dong-Hwan;Sohn, Young-Soo
    • Journal of Sensor Science and Technology
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    • v.19 no.6
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    • pp.456-461
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    • 2010
  • The detection of C-reactive protein(CRP) using self-assembled monolayer(SAM) was investigated by a portable surface plasmon resonance(SPR) sensor system. The CRP is a biomarker for the possible cardiovascular disease. The SAM was formed on gold(Au) surface to anchor the monoclonal antibody of CRP(anti-CRP) for detection of CRP. Sequence injection of the anti-CRP and bovine serum albumin(BSA) into the sensor system has been carried out immobilize the antibody and to prevent non-specific binding. The portable SPR system has two flow channels: one for the sample measurements and the other for the reference. The output SPR signal was increased with the injection of the anti-CRP, BSA and CRP due to binding of the proteins on the sensor chip. The valid output SPR signals was linearly related to the critical range of the CRP concentration. The experimental results showed the feasibility of the portable SPR system with newly developed SAM to diagnose a risk of the future cardiovascular events.

A coupled finite element/meshfreemoving boundary method for self-piercing riveting simulation

  • Cai, Wayne;Wang, Hui-Ping;Wu, C.T.
    • Interaction and multiscale mechanics
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    • v.6 no.2
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    • pp.257-270
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    • 2013
  • The use of lightweight materials has been steadily increasing in the automotive industry, and presents new challenges to material joining. Among many joining processes, self-piercing riveting (SPR) is particularly promising for joining lightweight materials (such as aluminum alloys) and dissimilar materials (such as steel to Al, and metal to polymer). However, to establish a process window for optimal joint performance, it often requires a long trial-and-error testing of the SPR process. This is because current state of the art in numerical analysis still cannot effectively resolve the problems of severe material distortion and separation in the SPR simulation. This paper presents a coupled meshfree/finite element with a moving boundary algorithm to overcome these numerical difficulties. The simulation results are compared with physical measurements to demonstrate the effectiveness of the present method.

Optimal Stiffness Design of Self-Piercing Riveting's C-Frame for Multimaterial Joining (다종소재 접합을 위한 SPR(Self-Piercing Riveting)용 C-프레임 강성 최적설계)

  • Shin, Chang-Yeul;Lee, Jae-Jin;Mun, Ji-Hun;Kwon, Soon-Deok;Yang, Min-Seok;Lee, Jae-Wook
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.20 no.5
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    • pp.76-84
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    • 2021
  • In this study, an optimal stiffness model of the C-frame, which was supporting the mold and tool load, was proposed to obtain quality self-piercing riveting (SPR) joining. First, the load path acting on the C-frame structure was identified using topology optimization. Then, a final suggested model was proposed based on the load path results. Stiffness and strength analyses were performed for a rivet pressing force of 7.3 [t] to compare the design performance of the final proposed model with that of the initial model. Moreover, to examine the reliability of continuous and repeated processes, vibration analysis was performed and the dynamic stiffness of the final proposed model was reviewed. Additionally, fatigue analysis was performed to ascertain the fatigue characteristics due to simple repetitive loading. Finally, stiffness test was performed for the final proposed model to verify the analysis results. The obtained results differed from the analysis result by 2.9%. Consequently, the performance of the final proposed model was superior to that of the initial model with respect to not only the SPR fastening quality but also the reliability of continuous and repetitive processes.

Detection of Escherichia coli O157:H7 Using Immunosensor Based on Surface Plasmon Resonance

  • Oh, Byung-Keun;Kim, Young-Kee;Bae, Young-Min;Lee, Won-Hong;Choi, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.780-786
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    • 2002
  • An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Escherichia coli O157:H7. A self-assembled protein C layer on a gold (Au) surface was fabricated by adsorbing the mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol at various molar ratios and by activating chemical binding between free amine (-$NH_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) in series. The formation of a self-assembled protein G layer on an Au substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of the self-assembled protein G layer on the Au substrate, monoclonal antibody (Mab) against E. coli O157:H7 which was immobilized on protein G, and bound E. coli O157:H7 extracts on Immobilized Mab against E. coii O157:H7 were performed by atomic force microscopy (AFM). The detection limit of the SPR-based immunosensor for E. coli O157:H7 was found to be about $10^4$ cells/ml.

Measurement of Alcohol Contents in Alcoholic Beverages Using a Surface Plasmon Resonance Sensor (표면 플라즈몬 공명 센서를 이용한 주류의 알코올 함량 측정)

  • 조용진;안병학;김철진;김종태
    • Journal of Biosystems Engineering
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    • v.29 no.4
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    • pp.323-328
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    • 2004
  • A surface plasmon resonance (SPR) sensing system, which belonged to a family of thin film refractometry-based sensors, was fabricated to measure the alcohol contents in alcoholic beverages. In this study, a SPR owe was obtained by a photodiode array with 128 pixels and transformed into a refractive index which was calculated by means of the first moment of values below a baseline of 0.85. When the sensing system was applied to Soju, Cheonju, Igwaduju and Tacju highly linear relationships between the refractive index by SPR and the contents of alcohol were obtained. That is, the coefficients of determination in their linear models were 0.992, 0.933, 0.918 and 0.954, respectively. Meanwhile, the effect of sample preparations on the accuracy of measurement was analyzed. As for Soju and Tacju, the best calibration equations were obtained when no sample preparation was applied. The best calibration models for measurement of Cheonju and Igwaduju were obtained through the samples filtered by $C_{18}$ disks.

Characteristics Analysis of Total Internal Reflection-based Dielectric Multi-layer Sensor Using Plasmonics Phenomena (플라즈모닉스 현상을 이용한 전반사 기반 다층 유전체 박막 센서의 특성 분석)

  • Kim, Hong-Seung;Lee, Tae-Kyeong;Kim, Doo-Gun;Jung, You-Ra;Oh, Geum-Yoon;Lee, Byeong-Hyeon;Ki, Hyun-Chul;Choi, Young-Wan
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.25 no.7
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    • pp.516-520
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    • 2012
  • In this paper, we have theoretically analyzed and designed a dielectric multi-layer sensor with a SPR (surface plasmon resonance) using analytical calculation and FDTD (finite difference time-domain) methods. The proposed structure is composed of periodic layer and thin metal film. It has many advantages. One of that is a high sensitivity of the SPR. Another is a high Q-factor of the characteristics in the PhC (photonic crystals) micro-cavity structure. The incident light has double resonance characteristics, because the filtered light by PhC structure, dielectric multi-layer, is met the thin metal film for SPR effect. We have also observed the change of resonance characteristics according to the variation of effective index on the metal film.