• Title/Summary/Keyword: positively charged

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Addition of an N-Terminal Poly-Glutamate Fusion Tag Improves Solubility and Production of Recombinant TAT-Cre Recombinase in Escherichia coli

  • Kim, A-Hyeon;Lee, Soohyun;Jeon, Suwon;Kim, Goon-Tae;Lee, Eun Jig;Kim, Daham;Kim, Younggyu;Park, Tae-Sik
    • Journal of Microbiology and Biotechnology
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    • v.30 no.1
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    • pp.109-117
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    • 2020
  • Cre recombinase is widely used to manipulate DNA sequences for both in vitro and in vivo research. Attachment of a trans-activator of transcription (TAT) sequence to Cre allows TAT-Cre to penetrate the cell membrane, and the addition of a nuclear localization signal (NLS) helps the enzyme to translocate into the nucleus. Since the yield of recombinant TAT-Cre is limited by formation of inclusion bodies, we hypothesized that the positively charged arginine-rich TAT sequence causes the inclusion body formation, whereas its neutralization by the addition of a negatively charged sequence improves solubility of the protein. To prove this, we neutralized the positively charged TAT sequence by proximally attaching a negatively charged poly-glutamate (E12) sequence. We found that the E12 tag improved the solubility and yield of E12-TAT-NLS-Cre (E12-TAT-Cre) compared with those of TAT-NLS-Cre (TAT-Cre) when expressed in E. coli. Furthermore, the growth of cells expressing E12-TAT-Cre was increased compared with that of the cells expressing TAT-Cre. Efficacy of the purified TAT-Cre was confirmed by a recombination test on a floxed plasmid in a cell-free system and 293 FT cells. Taken together, our results suggest that attachment of the E12 sequence to TAT-Cre improves its solubility during expression in E. coli (possibly by neutralizing the ionic-charge effects of the TAT sequence) and consequently increases the yield. This method can be applied to the production of transducible proteins for research and therapeutic purposes.

Regular Distribution of -OH Fragments on a Si (001)-c(4×2) Surface by Dissociation of Water Molecules (물 분자의 해리에 의한 Si (001)-c(4×2) 표면에서의 수산화기의 균일한 분포)

  • Lee, Soo-Kyung;Oh, Hyun-Chul;Kim, Dae-Hee;Jeong, Yong-Chan;Baek, Seung-Bin;Kim, Yeong-Cheol
    • Korean Journal of Materials Research
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    • v.20 no.9
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    • pp.457-462
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    • 2010
  • Adsorption of a water molecule on a Si (001) surface and its dissociation were studied using density functional theory to study the distribution of -OH fragments on the Si surface. The Si (001) surface was composed of Si dimers, which buckle in a zigzag pattern below the order-disorder transition temperature to reduce the surface energy. When a water molecule approached the Si surface, the O atom of the water molecule favored the down-buckled Si atom, and the H atom of the water molecule favored the up-buckled Si atom. This is explained by the attractions between the negatively charged O of the water and the positively charged down-buckled Si atom and between the positively charged H of the water and the negatively charged up-buckled Si atom. Following the adsorption of the first water molecule on the surface, a second water molecule adsorbed on either the inter-dimer or intra-dimer site of the Si dimer. The dipole-dipole interaction of the two adsorbed water molecules led to the formation of the water dimer, and the dissociation of the water molecules occurred easily below the order-disorder transition temperature. Therefore, the 1/2 monolayer of -OH on the water-terminated Si (001) surface shows a regular distribution. The results shed light on the atomic layer deposition process of alternate gate dielectric materials, such as $HfO_2$.

Programmed APTES and OTS Patterns for the Multi-Channel FET of Single-Walled Carbon Nanotubes (SWCNT 다중채널 FET용 표면 프로그램된 APTES와 OTS 패턴을 이용한 공정에 대한 연구)

  • Kim, Byung-Cheul;Kim, Joo-Yeon;An, Ho-Myoung
    • The Journal of Korea Institute of Information, Electronics, and Communication Technology
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    • v.8 no.1
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    • pp.37-44
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    • 2015
  • In this paper, we have investigated a selective assembly method of single-walled carbon nanotubes (SWCNTs) on a silicon substrate using only photolithographic process and then proposed a fabrication method of field effect transistors (FETs) using SWCNT-based patterns. The aminopropylethoxysilane (APTES) patterns, which are formed for positively charged surface molecular patterns, are utilized to assemble and align millions of SWCNTs and we can more effectively assemble on a silicon (Si) surface using this method than assembly processes using only the 1-octadecyltrichlorosilane (OTS). We investigated a selective assembly method of SWCNTs on a Si surface using surface-programmed APTES and OTS patterns and then a fabrication method of FETs. photoresist(PR) patterns were made using photolithographic process on the silicon dioxide (SiO2) grown Si substrate and the substrate was placed in the OTS solution (1:500 v/v in anhydrous hexane) to cover the bare SiO2 regions. After removing the PR, the substrate was placed in APTES solution to backfill the remaining SiO2 area. This surface-programmed substrate was placed into a SWCNT solution dispersed in dichlorobenzene. SWCNTs were attracted toward the positively charged molecular regions, and aligned along the APTES patterns. On the contrary, SWCNT were not assembled on the OTS patterns. In this process, positively charged surface molecular patterns are utilized to direct the assembly of negatively charged SWCNT on SiO2. As a result, the selectively assembled SWCNT channels can be obtained between two electrodes(source and drain electrodes). Finally, we can successfully fabricate SWCNT-based multi-channel FETs by using our self-assembled monolayer method.

Preparation of Positively and Negatively Charged Carbon Nanotube-Collagen Hydrogels with pH Sensitive Characteristic (양전하와 음전하를 띄며 pH 감응성인 카본나노튜브-콜라젠 Hydrogel의 합성)

  • Seo, Jae-Won;Shin, Ueon Sang
    • Journal of the Korean Chemical Society
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    • v.60 no.3
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    • pp.187-193
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    • 2016
  • In this study, preparation of positively and negatively charged carbon nanotube (CNT)-collagen (CG) hydrogels with pH sensitive characteristic was reported. The positive and negative characteristics of the prepared hydrogels were created by introduction of positively functionalized CNT-NH2 and negatively functionalized CNT-COOH, respectively, into the collagen hydrogel. The surface charge of CNTs (CNT-NH2 and CNT-COOH), CG and CNTs/CG hydrogels was measured by Zetasizer. The swelling ratios of CNT-NH2/CG and CNT-COOH/CG hydrogels in aqueous solution were checked by measuring of weight changes of the hydrogels in the range of pH 2~10. In detail, the positively charged CNT-NH2/CG hydrogel swelled up to 5% at pH 4 in comparison to the weight at pH 7, while the negatively charged CNT-COOH/CG hydrogel swelled up to 10% at pH 10. The prepared CNT-NH2/CG and CNT-COOH/CG hydrogels will be very useful as pH sensitive oral drug-delivering systems for gastrointestine (pH ~2) and small intestine (pH ~9), respectively.

A Study on the Streaming Electrification and Static Charge Elimination of Insulating Oil on Pipe Materials. (파이프 재질에 따른 절연유의 대전및재전에 관한 연구)

  • Kim, Y.W.;Cho, Y.K.;Shin, Y.D.;Jeong, H.D.;Lee, D.C.
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1993.05a
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    • pp.72-75
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    • 1993
  • The characteristic of streaming electrification and static charge diminution in Cu, Fe and Sus pipes were investigated. The Cu and Fe pipes are charged positively, whereas the Sus pipes are charged negatively. Electrification rate in Cu and Fe pipes increases with increasing a pipe length for no-charged insulating oil. The electrification rate in Cu and Fe pipes abruptly increase with increasing the temperature and flow rate of insulating oil but decrease in Sus pipes. The current flows as a direction of insulating oil in all pipes.

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Study on pH Sensor using Methylene Blue Adsorption and A Long-Period Optical Fiber Grating Pair

  • Jeon Young-Hee;Kwon Jae-Joong;Lee Byoung-Ho
    • Journal of the Optical Society of Korea
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    • v.10 no.1
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    • pp.28-32
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    • 2006
  • We propose a new pH-sensing scheme using a methylene blue adsorption on an optical fiber cladding surface. Interactions between the silica and hydroxyl ions of a base solution induce the surface of the silica negatively charged. The charged surface attracts the positively charged chromophores of methylene blue. As the pH of the solution is reduced, the electrostatic attraction will also be reduced. This electrostatic attraction can change the transmitted light intensity of the cladding mode, since the boundary condition changes. We also carried out a simulation to verify the effect from external refractive index change around a long-period fiber grating. Our results confirm that the wavelength shift by external refractive index change is negligible compared to the transmitted light intensity variation of the cladding mode. By using a long-period grating pair, we can detect the cladding mode transmittance variations. Experimentally, we showed the possibility of pH sensing in the $1.5{\mu}m$ infrared region.

Adhesion Force Analysis of Charged Particles for the E-paper (전자 종이용 하전 입자의 부착력 분석)

  • Kim, Seung-Taek;Kim, Hyung-Tae;Lee, Sang-Ho;Kim, Jong-Seok
    • Journal of the Semiconductor & Display Technology
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    • v.9 no.4
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    • pp.87-91
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    • 2010
  • Charged micro-particles are widely used as the key components for many electrical applications such as an e-paper, a touch panel, a printer toner and an electronic ink. Among them, the e-paper is an emerging reflective type display using the charged particles that has the advantages of the extremely low power consumption and sunlight readability. To create images on the e-paper, we confine black positively-charged and white negatively-charged particles between bottom and top electrodes and selectively apply the electric field. When the Coulomb force by an applied electric field is greater than the adhesion force between the charged particle and the electrode, the particles' transition happens resulting in the change of color between black and white. Therefore, the adhesion force is a very important factor for designing and estimating e-paper's operation. In this study, we constructed a basic model for particle's transition and an adhesion force equation describing particle's transition with three different forces: electrostatic image force, Van der Waals force and gravitational force. The simulation results showed that the gravitational force is negligible for the interesting range for the charge and the radius, and the adhesion force can be strongly dependent on the particle's charge and radius.

Layer-by-layer assembled graphene oxide films and barrier properties of thermally reduced graphene oxide membranes

  • Kim, Seon-Guk;Park, Ok-Kyung;Lee, Joong Hee;Ku, Bon-Cheol
    • Carbon letters
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    • v.14 no.4
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    • pp.247-250
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    • 2013
  • In this study, we present a facile method of fabricating graphene oxide (GO) films on the surface of polyimide (PI) via layer-by-layer (LBL) assembly of charged GO. The positively charged amino-phenyl functionalized GO (APGO) is alternatively complexed with the negatively charged GO through an electrostatic LBL assembly process. Furthermore, we investigated the water vapor transmission rate and oxygen transmission rate of the prepared (reduced GO $[rGO]/rAPGO)_{10}$ deposited PI film (rGO/rAPGO/PI) and pure PI film. The water vapor transmission rate of the GO and APGO-coated PI composite film was increased due to the intrinsically hydrophilic property of the charged composite films. However, the oxygen transmission rate was decreased from 220 to 78 $cm^3/m^2{\cdot}day{\cdot}atm$, due to the barrier effect of the graphene films on the PI surface. Since the proposed method allows for large-scale production of graphene films, it is considered to have potential for utilization in various applications.

Heterogeneous Electron Transfer at Polyoxometalate-modified Electrode Surfaces

  • Choi, Su-Hee;Seo, Bo-Ra;Kim, Jong-Won
    • Bulletin of the Korean Chemical Society
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    • v.31 no.1
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    • pp.104-111
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    • 2010
  • The heterogeneous electron transfer at $SiMo_{12}O_{40}^{4-}$ monolayers on GC, HOPG, and Au electrode surfaces are investigated using cyclic voltammetric and electrochemical impedance spectroscopic (EIS) methods. The electron transfer of negatively charged $Fe(CN)_6^{3-}$ species is retarded at $SiMo_{12}O_{40}^{4-}$-modified electrode surfaces, while that of positively charged $Ru(NH_3)_6^{3+}$species is accelerated at the modified surfaces. This is due to the electrostatic interactions between $SiMo_{12}O_{40}^{4-}$ layers on surfaces and charged redox species. The electron transfer kinetics of a neutral redox species, 1,1‘-ferrocenedimethanol (FDM), is not affected by the modification of electrode surfaces with $SiMo_{12}O_{40}^{4-}$, indicating the $SiMo_{12}O_{40}^{4-}$ monolayers do not impart barriers to electron transfer of neutral redox species. This is different from the case of thiolate SAMs which always add barriers to electron transfer. The effect of $SiMo_{12}O_{40}^{4-}$ layers on the electron transfer of charged redox species is dependent on the kind of electrodes, where HOPG surfaces exhibit marked effects. Possible mechanisms responsible for different electron transfer behaviors at $SiMo_{12}O_{40}^{4-}$ layers are proposed.

Effect of Bovine Serum Albumin on the Stability of Methotrexate-encapsulated Liposomes

  • Kim, Chong-Kook;Kim, Han-Sung;Lee, Beum-Jin;Han, Jeong-Hee
    • Archives of Pharmacal Research
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    • v.14 no.4
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    • pp.336-341
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    • 1991
  • The effect of bovine serum albumin (BSA) on the encapsulation efficiency and stability of liposomes containing methotrexate (MTX) having different surface charges and cholesterol contents were investigated. The encapsulation efficiency of MTX was lower and the release of MTX was faster by the addition of BSA. The leaking of MTX from lipid bilayer depends upon the BSA concentrations. These results may be derived from the interaction of BSA with lipid bilayers. The dynamic structural changes of BSA were monitored indirectly using circular dichroism spectra. Observed dynamic structural changes of BSA with liposomes are presumed to reflect the interaction of BSA with liposomes. Negatively charged liposomes have more strong interaction with BSA than neutral and positively charged liposomes. BSA attacks lipid bilayers whether it is at the inner or at the outer phase of lipid bilayer and induces leakage of entrapped MTX. Especially, negatively charged liposomes are more sensitive than others. The inclusion of cholesterol in the lipid layers inhibits the interaction of BSA with liposomes and shows protective effect against BSA-induced leakage of MTX. To endure the attacking of BSA liposomes as drug carriers should be made using cholesterol.

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