• Korean Journal of Materials Research
• /
• v.22 no.8
• /
• pp.433-438
• /
• 2012

A chemical process involves polymerization within microspheres, whereas a physical process involves the dispersion of polymer in a nonsolvent. Nano-sized monodisperse microspheres are usually prepared by chemical processes such as water-based emulsions, seed suspension polymerization, nonaqueous dispersion polymerization, and precipitation polymerizations. Polymerization was performed in a four-necked, separate-type flask equipped with a stirrer, a condenser, a nitrogen inlet, and a rubber stopper for adding the initiator with a syringe. Nitrogen was bubbled through the mixture of reagents for 1 hr. before elevating the temperature. Functional silane (3-mercaptopropyl)trimethoxysilane (MPTMS) was used for the modification of silica nanoparticles and the self-assembled monolayers obtained were characterized by X-ray photoelectron spectroscopy (XPS), laser scattering system (LSS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), elemental analysis (EA), and thermogravimetric analysis (TGA). In addition, polymer microspheres were polymerized by radical polymerization of ${\gamma}$-mercaptopropyl modified silica nanoparticles (MPSN) and acrylamide monomer via precipitation polymerization; then, their characteristics were investigated. From the elemental analysis results, it can be concluded that the conversion rate of acrylamide monomer was 93% and that polyacrylamide grafted to MPSN nanospheres via the radical precipitation polymerization with AAm in ethanol solvent. The microspheres were successfully polymerized by the 'graft from' method.

• Journal of Applied Biological Chemistry
• /
• v.58 no.3
• /
• pp.227-232
• /
• 2015

A microarray study has been employed to understand changes of gene expression in E. coli KD43162 resistant to ampicillin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, cefazolin, cefepime, aztreonam, imipenem, meropenem, gentamicin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, fosfomycin, and trimethoprim-sulfamethoxazole except for amikacin using disk diffusion assay. Using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF MS analyses, 36 kDa of outer membrane proteins (OMPs) was found to be deleted in the multidrug resistant E. coli KD 43162. Microarray analysis was used to determine up- and down-regulated genes in relation to multidrug resistant E. coli KD43162. Among the up-regulated genes, these genes were corresponded to express the proteins as penicillin-binding proteins (PBPs), tartronate semialdehyde reductase, ethanolamine utilization protein, shikimate kinase I, allantoinase, predicted SAM-dependent methyltransferase, L-glutamine: D-fructose-6-phosphate aminotransferase (GFAT), phospho-glucosamine mutase, predicted N-acetylmannosamine kinase, and predicted N-acetylmannosamine-6-P epimerase. Up-regulation of PBPs, one of primary target sites of antibiotics, might be responsible for the multidrug resistance in E. coli with increasing amount of target sites. Up-regulation of GFAT enzyme may be related to the up-regulation of PBPs because GFAT produces N-acetylglucosamine, a precursor of peptidoglycans. One of GFAT inhibitors, azaserine, showed a potent inhibition on the growth of E. coli KD43162. In conclusion, up-regulation of PBPs and GFATs with the loss of 36 kDa OMP refers the multidrug resistance in E. coli KD 43162.

• Applied Biological Chemistry
• /
• v.40 no.6
• /
• pp.546-550
• /
• 1997

This study was carried out to elucidate the biological characteristics of Rhizobia in biological nitrogen fixation system. The results of investigation were as follows; Polyacrylamide gel electrophoresis pattern of root lectin in the presence of SDS was ascertained electrophoretically and chromatographically. The purified root lectin formed immunoprecipitin line with anti lectin rabbit IgG. Root lectin, seed lectin and root exudate were tested for chemotactic ability. Chemotactic responses of RCR3407 and KCTC2422 toward root exudate were stronger than those of seed lectin and root lectin, but there didn't occur chemotactic responses of LPN100, not bound with seed lectin and that of LPN101, bound with seed lectin toward root exudate, root lectin and seed lectin. RCR3407, KCTC2422 and LPN-101, which nodulated with soybean, interacted with soybean lectin, but not with pea lectin. LPN-100, which was not nodulated with soybean, didn't interact with soybean lectin.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.50 no.1
• /
• pp.32-38
• /
• 2005

A long-term study initiated in 1989 at San-born Field, Columbia, Missouri, was designed to evaluate the affect of environmental factors, nitrogen application, and crop rotation on soybean (Glycine max [L.] Merr.) seed composition. Soybeans were grown as part of a four- year rotation which included corn (Zea maize L.), wheat (Triticum aestivum L.), and red clover (Trifolium pratense L.). Results from soil tests made prior to initiation of the study and subsequently every five years, were used to calculate application rates of nitrogen, phosphorus, and potassium necessary for target yield of pursuant crops. In the experimental design, nitrogen was applied to one-half of the plot on which the non-leguminous crop, either corn or wheat was grown. Analysis of soybean seed by near infrared reflectance spectroscopy collected over an 11-year period revealed a linear increase in protein and decrease in oil content. Application of nitrogen fertilizer to non-leguminous crops did not have an apparent effect on total protein or oil content of subsequent soybean crop. Analysis of soybean seed proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis in conjunction with computer­assisted densitometry revealed subtle changes in the accumulation of seed proteins. Immunoblot analysis using antibodies raised against the $\beta-subunit$ of $\beta-conglycinin$ showed a gradual increase in the accumulation of the 7S components during successive years of the experiment. A linear increase in temperature and decrease in rainfall was observed from the onset of data· collection. Higher temperatures during the growing season have been linked to increased protein and diminished oil content of soybean, thus changes observed in this study are possibly related to climatic conditions. However, crop rotation and subsequent changes in soil ecology may contribute to these observed changes in the seed composition.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.2
• /
• pp.239-246
• /
• 2005

Some characteristics of two forms of glucoamylase (glucan 1 A-$\alpha$-glucosidase, EC 3. 2. I. 3) purified from Aspergillus coreanus NR 15-1 were investigated. The enzymes were produced on a solid, uncooked wheat bran medium of A. coreanus NR 15-1 isolated from traditional Korean Nuruk. Two forms of glucoamylase, GA-I and GA-II, were purified to homogenity after 5.8-fold and 9.6-fold purification, respectively, judged by disc- and SDS-polyacrylamide gel electrophoresis. The molecular mass of GA-I and GA-II were estimated to be 62 kDa and 90 kDa by Sephadex G-1OO gel filtration, and 64 kDa and 91 kDa by SDS-polyacrylarnide gel electrophoresis, respectively. The optimum temperatures of GA-I and GA-II were 60$^circ$C and 65$^circ$C, respectively, and the optimum pH was 4.0. The activation energy (Ea value) of GA-I and GA-II was 11.66 kcal/mol and 12.09 kcal/mol, respectively, and the apparent Michaelis constants (K_{m}) of GA-I and GA-II for soluble starch were found to be 3.57 mg/ml and 6.25 mg/ml, respectively. Both enzymes were activated by 1 mM Mn^{2+} and Cu^{2+}, but were completely inhibited by 1 mM N­bromosuccinimide. The GA-II was weakly inhibited by 1 mM p-CMB, dithiothreitol, EDTA, and pyridoxal 5-phosphate, but GA-I was not inhibited by those compounds. Both enzymes had significant ability to digest raw wheat starch and raw rice starch, and hydrolysis rates of raw wheat starch by GA-I and GA-II were 7.8- and 7.3-fold higher than with soluble starch, respectively.

• Korean Journal of Environmental Biology
• /
• v.17 no.2
• /
• pp.209-215
• /
• 1999

Changes of malate dehydrogenase isozyme in oyster exposed to different temperature, pH and salinity were investigated by polyacrylamide gel electrophoresis. MDH isozyme in control group was separated into two bands on the positive side. In case of temperature and pH stress, MDH isozyme was separated into only one band after 12 hours exposure but two bands after 24, 48 hours exposure on the positive side. In case of salinity stress, after 12 hours exposure, MDH isozyme bands were separated into two bands in 5 ppt, 30 ppt and three bands in 10 ppt, 40 ppt concentration on the positive side. After 24 hours and 48 hours exposure case in salinity stress, MDH isozyme bands was separated into two bands on the positive side in all concentration. Activities of isozyme bands show their characteristics according to the condition of experiment. In conclusion, changes of MDH isozyme was a biochemical defense mechanism in oyster and result from effect of environmental stress to oyster.

• Journal of fish pathology
• /
• v.9 no.2
• /
• pp.177-183
• /
• 1996

A new RNA virus isolated from abnormally swimming behavior has caused mortalities in salmonid fish (Oh et al., 1995 a), A reverse transcriptase (RTase) activity of the virus was determined by using poly r(A) : oligo d(T) as templete : primer. This RTase activity was associated with virus particles of buoyant density of 1.16 g/ml. The virus particles in sucrose fractions were enveloped and were about 85 nm diameter with central electron-dense core. The brain and kidney samples of artificially infected fish showed RTase activity. Virus particle associated proteins about 120, 80, 65, 61, 48, 42, 35, 30, 25, 19, and 15 kDa were observed when examined by polyacrylamide gel electrophoresis analysis. This study showed the presence of a new retrovirus in salmonid fish, which tentatively called RVS (Retrovirus of salmonid).

• Journal of Embryo Transfer
• /
• v.25 no.4
• /
• pp.281-286
• /
• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.3
• /
• pp.525-531
• /
• 2005

From a genomic library of Lactobacillus reuteri ATCC 55739, one clone, NE347, carrying a pyrH gene encoding UMP kinase, was identified. pNE347 carried a 1.88 kb EcoRI fragment and the pyrH was located in the middle of the insert. pyrH ORF was 723 bp in size and capable of encoding UMP kinase composed of 240 amino acid residues. tsf encoding an elongation factor-Ts and frr encoding a ribosomal recycling factor were present upstream and downstream of pyrH, respectively. When introduced into E. coli KUR1244, a pyrH-negative strain, pNE347 restored the ability to grow at $42^{\circ}C$, indicating that pyrH from L. reuteri synthesized functional UMP kinase in E. coli. Northern blot experiment showed that pyrH and frr were cotranscribed as a 1.4 kb single transcript. pyrH was overexpressed in E. coli by using a pET26b(+) vector, and a major 25 kDa protein band appeared on SDS-polyacrylamide gel.

• Journal of Life Science
• /
• v.14 no.2
• /
• pp.309-314
• /
• 2004

Cyclic AMP receptor protein (CRP) is involved in the transcriptional regulation of more than 100 genes in E. coli. CRP dimer is converted into active form via the sequential conformation change of cAMP binding pocket, hinge region and HTH DNA binding motif by binding of cAMP. The temperature gradient gel electrophoresis (TGGE) was applied to CRP protein to know whether it was an efficient technique to study the conformational transitions and the thermal stability. TGGE showed the unfolding process of wild-type and S83G CRP proteins with the temperature gradient set from 29 to 71$^{\circ}C$ on nondenaturing polyacrylamide gel. Melting temperature (Tm) was 57$\pm$1 and 55$\pm$1$^{\circ}C$ for wild-type and S83G CRP, respectively in acidic buffer[89.8 mM Glycine and 24 mM Boric acid (pH 5.8)].