• Title/Summary/Keyword: molecular method

Search Result 3,663, Processing Time 0.026 seconds

Comparison of Ectopic Gene Expression Methods in Rat Neural Stem Cells

  • Kim, Woosuk;Kim, Ji Hyeon;Kong, Sun-Young;Park, Min-Hye;Sohn, Uy Dong;Kim, Hyun-Jung
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.17 no.1
    • /
    • pp.23-30
    • /
    • 2013
  • Neural stem cells (NSCs) have the ability to proliferate and differentiate into various types of cells that compose the nervous system. To study functions of genes in stem cell biology, genes or siRNAs need to be transfected. However, it is difficult to transfect ectopic genes into NSCs. Thus to identify the suitable method to achieve high transfection efficiency, we compared lipid transfection, electroporation, nucleofection and retroviral transduction. Among the methods that we tested, we found that nucleofection and retroviral transduction showed significantly increased transfection efficiency. In addition, with retroviral transduction of Ngn2 that is known to induce neurogenesis in various types of cells, we observed facilitated final cell division in rat NSCs. These data suggest that nucleofection and retroviral transduction provide high efficiency of gene delivery system to study functions of genes in rat NSCs.

Modulation of Poly($\beta-amino ester$) pH-Sensitive Polymers by Molecular Weight Control

  • Kim Min Sang;Lee Doo Sung;Choi Eun-Kyung;Park Heon-Joo;Kim Jin-Seok
    • Macromolecular Research
    • /
    • v.13 no.2
    • /
    • pp.147-151
    • /
    • 2005
  • The main objective of this study was to modulate pH-sensitive polymers (poly($\beta-amino esters$)) by controlling their molecular weight during their synthesis. These pH-sensitive and biodegradable polymers were synthesized by Michael-type step polymerization. 1,4-Butane diol diacrylate was used as the unsaturated carbonyl compound and piperazine as the nucleophilic compound. Various molecular weight polymers were obtained by varying the mol ratio of piperazine/1,4-butane diol diacrylate. The synthesized polymers were characterized by $^{1}H-NMR$ and their molecular weights were measured by gel permeation chromatography(GPC). The dependence of the molecular weight on the mol ratio was evaluated by the titration method. Also, the pH dependent turbidity of the polymers was determined by UV-Vis spectrophotometry. This pH dependent property of the polymers could be very useful for preparing drug carriers that are sensitive to pH.

Effects of Molecular Weight of PC on Mechanical Properties of PC/ABS Blends using High-Shear Rate Processing

  • Lee, Eun Ju;Park, Hee Jung;Kim, Se Mi;Lee, Seung Goo;Lee, Kee Yoon
    • Korean Chemical Engineering Research
    • /
    • v.56 no.3
    • /
    • pp.343-348
    • /
    • 2018
  • Each of the two polycarbonates (PC) of different molecular weights was blended with acrylonitrile-butadiene-styrene (ABS) under high-shear rate processing to afford PC/ABS. Sizes of ABS dispersed phases and mechanical properties of PC/ABS blends were investigated and high-shear rate processing of PC/ABS was carried out by changing screw speed and processing time. Prepared specimens were examined by scanning electron microscope (SEM) to observe morphology changes. Sizes of ABS dispersed phases in PC/ABS blends were observed to decrease gradually as screw speeds increased. Tensile strengths and elongations of specimens were investigated by universal testing method (UTM) to study the influence of molecular weight of PC exerting on PC/ABS blends. As a result, PC1/ABS blends (PC1: higher molecular weight PC) exhibited more strengthened properties than PC2/ABS (PC2: lower molecular weight PC). The tensile strength of PC1/ABS showed an increasing tendency when the screw speed increased, and the elongation did not show a significant decrease, but increased slightly with increasing shear time at a constant screw speed of 1000 rpm.

Structural Design and Characterization of a Channel-forming Peptide

  • Krittanai, Chartchai;Panyim, Sakol
    • BMB Reports
    • /
    • v.37 no.4
    • /
    • pp.460-465
    • /
    • 2004
  • A 16-residue polypeptide model with the sequence acetyl-YALSLAATLLKEAASL-OH was derived by rational de novo peptide design. The designed sequence consists of amino acid residues with high propensity to adopt an alpha helical conformation, and sequential order was arranged to produce an amphipathic surface. The designed sequence was chemically synthesized using a solid-phase method and the polypeptide was purified by reverse-phase liquid chromatography. Molecular mass analysis by electro-spray ionization mass spectroscopy confirmed the correct designed sequence. Structural characterization by circular dichroism spectroscopy demonstrated that the peptide adopts the expected alpha helical conformation in 50% acetonitrile solution. Liposome binding assay using Small Unilamellar Vesicle (SUV) showed a marked release of entrapped glucose by interaction between the lipid membrane and the tested peptide. The channel-forming activity of the peptide was revealed by a planar lipid bilayer experiment. An analysis of the conducting current at various applied potentials suggested that the peptide forms a cationic ion channel with an intrinsic conductance of 188 pS. These results demonstrate that a simple rational de novo design can be successfully employed to create short peptides with desired structures and functions.

Enhanced Expression of High-affinity Iron Transporters via H-ferritin Production in Yeast

  • Kim, Kyung-Suk;Chang, Yu-Jung;Chung, Yun-Jo;Park, Chung-Ung;Seo, Hyang-Yim
    • BMB Reports
    • /
    • v.40 no.1
    • /
    • pp.82-87
    • /
    • 2007
  • Our heterologous expression system of the human ferritin H-chain gene (hfH) allowed us to characterize the cellular effects of ferritin in yeasts. The recombinant Saccharomyces cerevisiae (YGH2) evidenced impaired growth as compared to the control, which was correlated with ferritin expression and with the formation of core minerals. Growth was recovered via the administration of iron supplements. The modification of cellular iron metabolism, which involved the increased expression of high-affinity iron transport genes (FET3 and FTR1), was detected via Northern blot analysis. The findings may provide some evidence of cytosolic iron deficiency, as the genes were expressed transcriptionally under iron-deficient conditions. According to our results examining reactive oxygen species (ROS) generation via the fluorescence method, the ROS levels in YGH2 were decreased compared to the control. It suggests that the expression of active H-ferritins reduced the content of free iron in yeast. Therefore, present results may provide new insights into the regulatory network and pathways inherent to iron depletion conditions.

Molecular Dynamics Study on Behaviors of Liquid Cluster with Shape and Temperature of Nano-Structure Substrate (나노구조기판의 형상 및 온도변화에 따른 액체 클러스터의 거동에 대한 분자동역학적 연구)

  • Ko, Sun-Mi;Jeong, Heung-Cheol;Shibahara, Masahiko;Choi, Gyung-Min;Kim, Duck-Jool
    • Journal of ILASS-Korea
    • /
    • v.13 no.1
    • /
    • pp.34-41
    • /
    • 2008
  • Molecular dynamic simulations have been carried out to study the effect of the nano-structure substrate and its temperature on cluster laminating. The interaction between substrate molecules and liquid molecules was modeled in the molecular scale and simulated by the molecular dynamics method in order to understand behaviors of the liquid cluster on nano-structure substrate. In the present model, the Lennard-Jones potential is applied to mono-atomic molecules of argon as liquid and platinum as nano-structure substrate to perform simulations of molecular dynamics. The effect of wettability on a substrate was investigated for the various beta of Lennard-Jones potential. The behavior of the liquid cluster and nano-structure substrate depends on interface wettability and function of molecules force, such as attraction and repulsion, in the collision progress. Furthermore, nano-structure substrate temperature and beta of Lennard-Jones potential have effect on the accumulation ratio. These results of simulation will be the foundation of coating application technology for micro fabrication manufacturing.

  • PDF

Development of a Rapid Molecular Detection Marker for Colletotrichum species with AFLP

  • Eom, Seung-Hee;Kim, Kwon-Jong;Jung, Hee-Sun;Lee, Sang-Pyo;Lee, Youn-Su
    • Mycobiology
    • /
    • v.32 no.3
    • /
    • pp.123-127
    • /
    • 2004
  • Sweet persimmons have been increasingly cultivated in the southern part of Korea. However, anthracnose disease caused by Colletotrichum species is one of the major hindrances in cultivation and productions. In this study, we used polymerase chain reaction(PCR) to detect Colletotrichum species with the AFLP(amplified fragment length polymorphism) method. In AFLP, we used E3(5'-GACTGCGTACCAATTCTA-3') and M1(5'-GATGAGTCCTGAGTAACAG-3') primer combination and, as a result, 262 bp segment was observed in Colletotrichum species only. Specific PCR primers were designed from the sequence data and used to detect the presence of the fungus in genomic DNA isolated from symptomless sweet persimmon plants. Based on sequence data for specific segments, Co.B1(5'-GAGAGAGTAGAATTGCGCTG-3') and Co.B2(5'-CTACCATTCTTCTA GGTGGG-3') were designed to detect Colletotrichum species. The 220 bp segment was observed in Colletotrichum species only, but not in other fungal and bacterial isolates.

On Some Changes in Polymer Blend Topological and Molecular Structures Resulted from Processing

  • Jurkowski, B.;Jurkowska, B.;Nah, C.
    • Elastomers and Composites
    • /
    • v.37 no.4
    • /
    • pp.234-243
    • /
    • 2002
  • A general scheme of a rubber structure is proposed. Using the thermomechanical method(TMA), some changes in the molecular and topological structures for uncured and cured, and unfilled and filled rubbers during processing are shown. In our investigations as region it is understood a complex structure, which is expressed at the thermomechanical curve(TMC) as a zone differed from others in thermal expansion properties. This zone is between the noticed temperatures of relaxation transitions, usually on the level like those determined by DMTA at 1Hz. These regions, which shares, are not stable, and differ in molecular-weight distribution(MWD) of chain fragments between the junctions. Differences in dynamics of the formation of the molecular and topological structures of a vulcanizate are dependent on the rubber formulation, mixing technology and curing time. Some of characteristics of these regions correlate with mechanical properties of vulcanizates what is shown for NR rubbers containing ENR or CPE as a polymeric additive. It is well known that the state of order influences diffusivity of low-molecular substances into the polymer matrix. Because of this, the two topological amorphous regions should influence the distribution of the ingredients and resulting in rubber compounds' heterogeneity, and related properties of cured rubber. Investigation of this problem is expected to be, in the future, one of the essential factors in determining further improvement of polymeric materials properties by compounding with additives and in reprocessing of rubber scrap.

Rapid Agrobacterium-mediated genetic rice transformation method using liquid media (액체배양을 이용한 단기 벼 형질전환 방법)

  • Yang, Dae-Hwa;Chang, Ahn-Cheol;Ahn, Il-Pyung;Kim, Hae-Jung;Kim, Dong-Hern;Lee, Hyo-Yeon;Suh, Seok Cheol
    • Journal of Plant Biotechnology
    • /
    • v.40 no.1
    • /
    • pp.37-42
    • /
    • 2013
  • Rice is one of the most important cereal crops as a model plant for functional genomics of monocotyledons and usually transformed using Agrobacterium tumefaciens. However, the transformation's process using previous method is still time consuming and uneconomical, low efficiency. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the infection and co-cultivation, Agrobacterium elimination, infected calli's selection steps using liquid media. We directly inoculated Agrobacterium containing a ZjLsL gene under the control of constitutive promoter into the 1- to 3-week-old rice calli derived from mature seeds. After 3 days of co-cultivation, the infected calli were transferred onto liquid media of Agrobacterium elimination and calli's selection for 3 days. The calli were transferred to calli's growth solid media for 14 days and then the calli transferred to shoot induction and root induction media. Putative transformants were initially selected on the medium containing phosphinothricin, and the PAT protein verified by PAT strip test. This method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

3D Printed Flexible Cathode Based on Cu-EDTA that Prepared by Molecular Precursor Method and Microwave Processing for Electrochemical Machining

  • Yan, Binggong;Song, Xuan;Tian, Zhao;Huang, Xiaodi;Jiang, Kaiyong
    • Journal of Electrochemical Science and Technology
    • /
    • v.11 no.2
    • /
    • pp.180-186
    • /
    • 2020
  • In this work, a metal-ligand solution (Cu-EDTA) was prepared based on the molecular precursor method and the solution was spin-coated onto 3D printed flexible photosensitive resin sheets. After being processed by microwave, a laser with a wavelength of 355 nm was utilized to scan the spin-coated sheets and then the sheets were immersed in an electroless copper plating solution to deposit copper wires. With the help of microwave processing, the adhesion between copper wires and substrate was improved which should result from the increase of roughness, decrease of contact angle and the consistent orientation of coated film according to the results of 3D profilometer and SEM. XPS results showed that copper seeds formed after laser scanning. Using the 3D printed flexible sheets as cathode and galvanized iron as anode, electrochemical machining was conducted.