• Title/Summary/Keyword: precursor signal

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Flow Field Change before Onset of Flow Separation

  • Hasegawa, Hiroaki;Sugawara, Takeru
    • International Journal of Fluid Machinery and Systems
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    • v.2 no.3
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    • pp.215-222
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    • 2009
  • Jets issuing through small holes in a wall into a freestream has proven effective in the control of flow separation. This technique is known as the vortex generator jet (VGJs) method. If a precursor signal of separation is found, the separation control system using VGJs can be operated just before the onset of separation and the flow field with no separation is always attained. In this study, we measured the flow field and the wall static pressure in a two-dimensional diffuser to find a precursor signal of flow separation. The streamwise velocity measurements were carried out in the separated shear layer and spectral analysis was applied to the velocity fluctuations at some angles with respect to the diffuser. The pattern of peaks in the spectral analysis changes as the divergence angle increases over the angle of which the whole separation occurs. This change in the spectral pattern is related to the enhancement of the growth of shear layer vortices and appears just before the onset of separation. Therefore, the growth of shear layer vortices can be regarded as a precursor signal to flow separation.

Toxicity of Tomato Spotted Wilt Virus Glycoprotein Signal Peptide and Promoter Activity of th 5' UTR

  • Park, Tae-Jin;Kim, Sun-Chang;Thomas L. German
    • The Plant Pathology Journal
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    • v.15 no.6
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    • pp.313-318
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    • 1999
  • Cloning of the 5'untranslated region (5' UTR) and Nterminus of the glycoprotein precursor (G2G1) open reading frame of tomato spotted wilt virus has been problematic, possibly because of the toxicity of a signal peptide at the beginning of th G2G1 protein precursor. The toxicity of the signal peptide to bacterial growth and the reason for the expression of the peptide gene in Escherichia coli were investigated by cloning the 5' UTR and the signal peptide sequence separately. Cells transformed with the plasmid containing both the first 30 amino acids of the glycoprotein and the 5' UTR showed a severe growth inhibition whereas transformants harboring either the plasmid with the signal sequence or the 5'UTR alone did not show any ingibition. An E. coli promoter-like sequence was found in the 5'UTR and tis promoter acivity was confirmed with a promoter-less GUS gene cloned downstream of the 5'UTR. In the cloning of the Tomato spotted wilt virus (TSWV) glycoprotein G2G1 open reading frame all the recovered plasmids contained stop codons in the signal sequence region. However, clones containing no stop codon were recovered when the signal sequence and the 5'UTR were cloned separately.

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High Level Expression of a Protein Precursor for Functional Studies

  • Gathmann, Sven;Rupprecht, Eva;Schneider, Dirk
    • BMB Reports
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    • v.39 no.6
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    • pp.717-721
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    • 2006
  • In vitro analyses of type I signal peptidase activities require protein precursors as substrates. Usually, these pre-proteins are expressed in vitro and cleavage of the signal sequence is followed by SDS polyacrylamide gel electrophoresis coupled with autoradiography. Radioactive amino acids have to be incorporated in the expressed protein, since the amount of the in vitro expressed protein is usually very low and processing of the signal peptide cannot be followed by SDS polyacrylamide gel electrophoresis alone. Here we describe a rapid and simple method to express large amounts of a protein precursor in E. coli. We have analyzed the effect of ionophors as well as of azide on the accumulation of expressed protein precursors. Azide blocks the function of SecA and the ionophors dissipate the electrochemical gradient across the cytoplasmic membrane of E. coli. Addition of azide ions resulted in the formation of inclusion bodies, highly enriched with pre-apo-plastocyanine. Plastocyanine is a soluble copper protein, which can be found in the periplasmic space of cyanobacteria as well as in the thylakoid lumen of cyanobacteria and chloroplasts, and the pre-protein contains a cleavable signal sequence at its N-terminus. After purification of cyanobacterial pre-apo-plastocyanine, its signal sequence can be cleaved off by the E. coli signal peptidase, and protein processing was followed on Coomassie stained SDS polyacrylamide gels. We are optimistic that the presented method can be further developed and applied.

Precursor and Substituent Effects on the Alternate Adsorption of Polycation and Anionic Dyes as Studied by Second Harmonic Generation and Absorption Spectroscopies

  • Yamada, Sunao;Niidome, Yasuro;Hinoguchi, Takeshi
    • Journal of Photoscience
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    • v.7 no.1
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    • pp.9-13
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    • 2000
  • An alternately assembled polycation-anionic dye film was prepared on a precursor assembly of poly(allylamine)hydrochloride (PAH) and poly(sodium styrenesulfonate) (PSS) or of poly (djallyldimethyl)ammonium chloride (PDDA) and PSS. The (PAH/PSS) precursor assembly gave better adsorptivity to the anionic dye than the (PDDA/PSS) one. Four kinds of anionic dyes (EB-T, AR-17 AR-18, AR-27) with different numbers of sulfonate substituents were compared. The extent of dye assembling was higher for the dye with a smaller number of sufonate substituents. Second harmonic generation (SHG) was clearly observed from the films. The SHG signal and the absorption intensity correlated well up to four polycation/dye bilayers, while further assembly did not increase the SHG signal appreciably. Second order nonlinear coefficients for ten bilayer assemblies were the order of pm/V.

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Development of Thermal Precursor DB for Partial Disconnection and Poor Contact on Electrical Wire (배선에서의 반단선 및 접촉불량에 대한 열적 전조 DB구축)

  • Kim, Doo-Hyun;Kim, Sung-Chul;Lee, Jong-Ho;Kang, Dong-Kyu
    • Journal of the Korean Society of Safety
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    • v.24 no.2
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    • pp.30-36
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    • 2009
  • This paper aims at the precursor analysis and DB development of electrical fires based on thermal and current signals for partial disconnection and poor contact on electric wires through experiments and simulations. Also, DB system required for developing the precursor DB with these data was studied and designed. Firstly, in case of partial disconnection, characteristics were derived and analyzed by experiment and electrical-thermal finite element method(Flux 3D) on the model wires which consist of VCTF and IV electric wires. Based on the characteristics, About 351 partial disconnection precursor patterns were generated by the thermal analysis for electric wire according to deterioration time under normal state and 200% overload state of rated current. Secondly, in order to develop poor contact precursor patterns, temperature value and the current signal were considered. In simulating the poor contact situation on connector area of MCCB, connection torque was changed. Through the experimental analysis, about 251 poor contact precursor patterns were generated. Finally, Using thermal precursor patterns obtained by partial disconnection and poor contact, electrical fire thermal precursor DB was developed.

Development of H/W and S/W for Detecting Electrical Fire Precursor Signal on Electrical Wirings (배선에서 전기화재 전조신호 검출을 위한 H/W 및 S/W 구축)

  • Kim, Sung-Chul;Kim, Doo-Hyun
    • Journal of the Korean Society of Safety
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    • v.24 no.3
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    • pp.13-18
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    • 2009
  • This paper is purposed to develop DAQ H/W, S/W and DB which can be used in developing electrical fire alarm system or in analyzing electrical fire cause, by detecting and monitoring precursor signals which have high possibility leading to electrical fire on electrical distribution wires. In this paper, developed was DAQ H/W adopting the C8051FXXX CPU which can analyze the measurement signals of current and voltage in electrical distribution wires, other CPU was investigated in view of the best digital sampling rate on the basis of previous researches for electrical fire alarm system. Also, the S/W which can interface with DAQ H/W's communication protocol and can be applied for electrical fire causes analysis, are embodied by LabVIEW. The combined DAQ H/W and S/W could analyze efficiently normal as well as abnormal electrical signals such as RMS value, instantaneous value of current and voltage, frequency signals etc, on the electrical wires. Also, DB system was constructed for recording various analysis results for precursor signals including voltage and current signals. The results by simulator and experiment showed that the suggested scheme with DAQ H/W, S/W and DB in this paper has high usability.

A Comparison of Three Dimensional Structures of Biosynthesized Preproinsulin and Insulin Using NMR

  • Oh, Mi-Na;Mok, K.-Hun;Lim, Yoong-Ho
    • Applied Biological Chemistry
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    • v.41 no.8
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    • pp.572-577
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    • 1998
  • The solution conformation of the human insulin precursor, preproinsulin, is described in terms of NMR spectral data. NMR experiments were performed on preproinsulin, whose structure was compared with the NMR structure of native human insulin. Despite the presence of the C-peptide and/or the signal peptide, secondary structure analyses indicate that the native structures of the A and B chains are well conserved even in preproinsulin. The observed relative robustness of the native structure in precursor forms permits further protein engineering experiments where the C-peptide or N-terminal signal sequence can be altered for the purpose of increasing expression or purification yields when producing recombinant human insulin.

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Fed-batch Culture of Recombinant E.coli for the Production of Penicillin G Amidase (Penicillin G Amidase생산을 위한 재조합 대장균의 유가배양에 관한 연구)

  • Lee, Sang-Mahn
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.314-319
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    • 2008
  • Penicillin G amidase (PGA, benzylpenicillinaminohydrolase, EC 3.5.1.11) is industrially important enzyme which converts penicillin G to 6-aminopenicillanic acid (6-APA) and phenylacetic acid (PAA). The PGA in E. coli ATCC 11105 is secreted into the periplasm after removing signal sequences and becomes heterodimer which composed of two subunits, small subunit (24 kDa) and large subunit (65 kDa). In this study, the PGA gene was obtained from E. coli ATCC 11105 using PCR (polymerase chain reaction) technique. The active PGA was successfully secreated into periplasm in E. coli BL2 1(DE3) harboring pET-pga plasmid. The optimized fed-batch fermentation, consisting of a three-step shift of culture temperature from $37^{\circ}C$ to $22^{\circ}C$, gave a productivity of 19.6 U/mL with a cell growth of 62 O.D. at 600 nm.

Preparation of Carbon Films from Polyacrylonitrile@Lignin Composites, and Their Electrical Properties and Adsorption Behavior (폴리아크릴로나이트릴/리그닌 복합소재로부터 생성된 탄소 필름의 전기적 성질 및 흡착 성능)

  • Joonwon Bae
    • Applied Chemistry for Engineering
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    • v.34 no.2
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    • pp.106-110
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    • 2023
  • Lignin is compatible with various polymeric materials and useful as a carbon precursor. In this work, carbon monolith films were produced from polyacrylonitrile (PAN)@lignin precursor films by a controlled carbonization cycle. In addition, their morphological features, electrical properties, and adsorption behavior were analyzed and compared with those of carbonized PAN films. The successful formation of PAN@lignin precursor was confirmed by Fourier-transform infrared (FT-IR) spectroscopy. SEM was used to examine the morphology of precursor and carbonized films, revealing that both precursor and carbonized films retained structural stability following carbonization. A trace of lignin in the carbonized films was also found. The pore structure of the carbonized PAN@lignin film was measured using the BET method, indicating the formation of fairly uniform pores. The electrical properties were also analyzed to obtain the Ohmic relation, which demonstrated that the electrical signal was influenced by incoming materials. Finally, the carbonized PAN@lignin films were useful as adsorbents to remove metal ions. This study provides important information for future initiatives in relevant research fields.

Cloning and Expression of a Alkaline Protease from Bacillus clausii I-52 (Bacillus clausii I-52로부터 alkaline protease 유전자의 클로닝 및 발현)

  • Joo, Han-Seung;Choi, Jang Won
    • Journal of agriculture & life science
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    • v.45 no.6
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    • pp.201-212
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    • 2011
  • The alkaline protease gene was cloned from a halo-tolerant alkalophilic Bacillus clausii I-52 isolated from the heavily polluted tidal mud flat of West Sea in Inchon Korea, which produced a strong extracellular alkaline protease (BCAP). Based on the full genome sequence of Bacillus subtilis, PCR primers were designed to allow for the amplification and cloning of the intact pro-BCAP gene including promoter region. The full-length gene consists of 1,143 bp and encodes 381 amino acids, which includes 29 residues of a putative signal peptide and an additional 77-amino-acid propeptide at its N-terminus. The mature BCAP deduced from the nucleotide sequence consists of 275 amino acids with a N-terminal amino acid of Ala, and a relative molecular weight and pI value was 27698.7 Da and 6.3, respectively. The amino acid sequence shares the highest similarity (99%) to the nattokinase precursor from B. subtilis and subtilisin E precursor from B. subtilis BSn5. The substrate specificity indicated that the recombinant BCAP could hydrolyze efficiently the synthetic substrate, N-Suc-Ala-Ala-Pro-Phe-pNA,and did not hydrolyze the substrates with basic amino acids at the P1 site. The recombinant BCAP was strongly inhibited by typical serine protease inhibitor, PMSF, indicating that BCAP is a member of the serine proteases.