• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 1995.06b
• /
• pp.97-117
• /
• 1995

Copper resistance in Pseudomonas syringae pv. tomato is determined by copper-resistance operon (cop) on a highly conserved 35 kilobase plasmid. Copper-resistant strains of Pseudomonas syringae containing the cop operon accumulate copper and develop blue clonies on copper-containing media. The protein products of the copper-resistance operon were characterized to provide an understanding of the copper-resistance mechanism and its relationship to copper accumulation. The Cop proteins CopA (72 kDa), CopB (39 kDa), and CopC (12 kDa) were produced only under copper induction. CopA and CopC were periplasmic proteins and CopB was an outer membrane protein. Leader peptide sequences of CopA, CopB, and CopC were confirmed by amino-terminal peptide sequencing. CopA, CopB, and CopC were purified from strain PT23.2, and their copper contents were determined. One molecule of CopA bound 10.9${\pm}$1.2 atoms of copper and one molecule of CopC bound 0.6${\pm}$0.1 atom of copper. P. syringae cells containing copCD or copBCD cloned behind the lac promoter were hypersensitive to copper. The CopD (32 kDa), a probable inner membrane protein, function in copper uptake with CopC. The Cop proteins apparently mediate sequestration of copper outside of the cytoplasm as a copper-resistance mechanism.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.37 no.1
• /
• pp.117-123
• /
• 2008

The protein profiles of domestic potato cultivars were evaluated for total protein determination, amino acid composition, SDS-PAGE analysis and scanning densitometry. There were statistically significant differences in the levels of amino acids among potato cultivars. Total nitrogen amount was also significantly different among cultivars, ranging from 1.27 to 1.64%. SDS-PAGE analysis showed that there were significant differences in the content of major potato proteins such as papatin (40 kDa), trypsin inhibitor (20 kDa) and protease inhibitor (15 kDa) among cultivars (p<0.05). The amount of papatin among cultivars with a range of 22.16 to 25.81 mg/g d.w. was higher in Jopung, Shepody and Superior, whereas the amount of protease inhibitors including 15 kDa and 20 kDa was the highest in Jopung (37.0%). The Shepody contains the highest amount of papatin (25.8%) and the lowest of trypsin inhibitor (5.22%). Thus, it is suggested that Shepody is the most desirable cultivar for better nutrition based on the protein profile.

• Korean journal of applied entomology
• /
• v.36 no.4
• /
• pp.331-340
• /
• 1997

Hyalophora cecropia attacin-like antibacterial gene was isolated from Bombyx mori induced with nonpathogenic bacteria. It was expressed in Spodopfera frugiperda 9 (Sf9 cells using baculovirus expression vector system (BEVS), and examined its antibacterial activity. With a cDNA library constructed from fifthinstar B. mori injected with Escherichia coli(4 X IOhcellsllarva), differential screening was performed using naive and induced mRNA probes. BmInc6 clone was screened by partial nucleotide sequence and GenBank database analysis. A complete nucleotide sequence of Bmlnc6 cDNA was determined (GenBank, AF005384). Its insert size was 852 bp and had open reading frame that started translation at position 35 and stopped at 679. And its putative polyadenylational signal existed at 812 bp. The number of amino acid deduced from Bmlnc6 cDNA was 214 and hydropathy analysis showed that this peptide was hydrophilic. This peptide deduced by BmInc6 was named nuecin. When the nuecin gene was expressed in Sf9 cells using BEVS, about 950 bp of the transcripts was detected. In addition, SDS-PAGE analysis showed that the molecular weights of intracellular expressed protein and the mature protein secreted to culture media were approximately 23 and 20 kDa, respectively. The antibacterial activity of nuecin against E. coli and Bacillus subtilis was significantly high, demonstrating that nuecin had a wider antibacterial spectrum with gram negative and positive bacteria than attacin.

• Archives of Pharmacal Research
• /
• v.18 no.5
• /
• pp.301-307
• /
• 1995

In the particulate fraction obtained from PKC-down regulated K562 cells by treatment for 24 h with 200nM TPA, phosphorylation of two proteins with molecular weight, 100 kDa and 23 kDa (designated p100 and p23, respectvely) was depleted and addition of exogenous purified PKC to this fraction failed to testore their phosphorylation. However, in the soluble fraction, all of phosphoproteins abolished by long-term treatment with TPA were restored by exogenously added PKC. Phosphorylation of two proteins was increased by short-term tretment (20 min), and diminished with the persistant exposure to TPA as well as at a concentration as low as 1nM. When K562 cells were treated with 1 nM and 200 nM TPA for 24 h, phosphorylation of p100 was restored with or without exogenous PKC on 2-3day and 6day after removal of treated TPA, respectively. Two-dimensional electrophoresis of phosphoproteins revealed that phosphorylated p100 (pl=5.9) and p66 species were completely absent from the particulate fraction of K562 cells treated with 200nM TPA for 24 h. These results suggest that the interaction of sensitive endogenous substrates, p100 and p23 with the plasma membrane might be regulated by PKC-down regulation without displacement to the cytosol and the interaction of p100 with the membrane might be reveersible.

• Parasites, Hosts and Diseases
• /
• v.29 no.2
• /
• pp.113-120
• /
• 1991

When the component proteins in crude saline extract of 13-week old adult Paragonimus westermani were observed by non-denaturing discontinuous-polyacrylamide gel electrophoresis (Disc-PAGE), 8 distinct bands were clearly recognized. Molecular weight (MW) of each band protein, numbered in sequence from cathodal side which appeared in 10% separating gel, was measured first by Ferguson plot utilizing different gel concentrations from 10% to 4.5%. MW of band 1 Protein (known as egg Protein) was 440 kDa. And MW of other band Proteins were: 386 kDa in band 2, 17.4 kDa in band 3, 17kDa in band 4, 14.3 kDa in band 5, 46 kDa in band 6, 38 kDa in band 7 and 23 kDa in band 8. When the proteins in the crude extract were separated into fractions by molecular sieve chromatography through 1.6 (Φ)×70cm sired Sephacryl 5-300 Superane column and revisualized by Disc-PAGE in 8% gel, the sequence of fluted proteins was band 1, band 2, band 6, band 7 and bands 3,4,5 and 8. This elusion profile confirmed MW of each band protein in the crude extract as measured by Ferguson plot.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.113.1-113
• /
• 2003

A potexvirus, Hosta virus X (HVX-Kr), causing mosaic and mottle symptoms was isolated from hosta plants (Hosta spp.), and its entire genome RNA sequence was determined. in Korea using cDNA library and RACE methods. The genome of HVX encodes five open reading frames coding for viral replicase, triple gene block (TGB), and viral coat protein (CP) from the 5'to 3' ends, which is a typical genome structure of potexviruses. The 3-terminal region of the virus includes the TGBI (26 kDa), TGB2 (13 kDa), TGB3 (8 kDa), and 23 kDa coat protein (CP) and the 3-nontranslated region (NTR). The CP gene of the type isolate of HVX (HVX-U) was amplified by RT-PCR and its nucleotide sequence was determined. The CPs of HVX-Kr and HVX-U had 100％ and 98.9％ identical amino acids and nucleotides, respectively. Most of the regions of the genome HVX had over 50％ nucleotide identical to other sequenced potexviruses. This is the first report of complete genome sequence information of HVX and molecular evidence supporting the virus as a distinct species of the genus Potexvirus.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.1 no.1
• /
• pp.19-23
• /
• 2000

A novel recombinant baculovirus Autographa californica nuclear polyhedrosis virus (ACNPV) producing the green fluorescent polyhedra was constructed and characterized. The recombinant virus was stably produced fluorescent polyhedra in the infected cells and the morphology of the polyhedra was nearly similar to that of wild-type AcNPV. For the production of the fluorescent polyhedral the green fluorescent protein (GFP) gene was introduced under the control of polyhedrin gene promoter of AcNPV by translational fusion in the front and back of intact polyhedrin gene. The recombinant baculovirus was named as CXEP, As expected, the 93 kDa fusion protein was expressed in the CXEP-infected cells. Interestingly, however, the cells infected with CXEP also showed a 33 kDa protein band as cells infected with wild-type AcNPV. The results of Southern blot analysis and plaque assay suggested that two types of baculoviruses expressing the GFP fusion protein or only native polyhedrin were formed through homologous recombination between two polyhedrin genes in the same orientation. Thus, this system can be applied for the production of recombinant polyhedra with foreign gene product of diverse interest.

• Journal of Animal Science and Technology
• /
• v.62 no.2
• /
• pp.121-140
• /
• 2020

Anaerobic fungi habitat in the gastrointestinal tract of foregut fermenters or hindgut fermenters and degrade fibrous plant biomass through the hydrolysis reactions with a wide variety of cellulolytic enzymes and physical penetration through fiber matrix with their rhizoids. To date, seventeen genera have been described in family Neocallimasticaceae, class Neocallimastigomycetes, phylum Neocallimastigomycota and one genus has been described in phylum Neocallimastigomycota. In National Center for Biotechnology Information (NCBI) database (DB), 23,830 nucleotide sequences and 59,512 protein sequences have been deposited and most of them were originated from Piromyces, Neocallimastix and Anaeromyces. Most of protein sequences (44,025) were acquired with PacBio next generation sequencing system. The whole genome sequences of Anaeromyces robustus, Neocallimastix californiae, Pecoramyces ruminantium, Piromyces finnis and Piromyces sp. E2 are available in Joint Genome Institute (JGI) database. According to the results of protein prediction, average Isoelectric points (pIs) were ranged from 5.88 (Anaeromyces) to 6.57 (Piromyces) and average molecular weights were ranged from 38.7 kDa (Orpinomyces) to 56.6 kDa (Piromyces). In Carbohydrate-Active enZYmes (CAZY) database, glycoside hydrolases (36), carbohydrate binding module (11), carbohydrate esterases (8), glycosyltransferase (5) and polysaccharide lyases (3) from anaerobic fungi were registered. During four decades, 1,031 research articles about anaerobic fungi were published and 444 and 719 articles were available in PubMed (PM) and PubMed Central (PMC) DB.

• Journal of Life Science
• /
• v.23 no.2
• /
• pp.299-305
• /
• 2013

Paenibacillus is a gram-positive, spore-forming aerobes that was previously classified as a Bacillus species. Paenibacillus sp. CK214 was highly motile on LB agar plates and showed typical colonial morphology of Paenibacillus. However, its motility was defective in the absence of glucose. Electron microscopic observation revealed that the cells of CK214 cultured on LB agar plates were peritrichously flagellated but not flagellated in the presence of glucose. Flagellar filaments were purified by centrifugation after shearing off from the CK214 cells with vigorous pipetting. The purified protein was composed of a single flagellin with an apparent molecular size of 29 kDa. Recognition of the protein by anti-Edwardsiella tarda flagellin protein antibody demonstrates that the protein is a flagellin protein. A decreased level of flagellin protein was detected in CK214 cells grown under glucose-supplemented media.

• The Journal of Pediatrics of Korean Medicine
• /
• v.14 no.2
• /
• pp.33-46
• /
• 2000

It has long been known that Jahage(紫河車) extracts of Placenta hominis are effective for immunological and vascular diseases in human body and thus, was used a major constituent of traditional oriental medicines. From full-term human placenta, we have purified a new type anticoagulant protein, PP27, using different chromatographic techniques of a phenyl TSK gel 650M column, DEAE, HA and Mono-Q columns. PP27 showed single band on SDS-PAGE with a molecular mass (Mr) of 27 kDa under denaturing conditions and a calibrated Sepharose 4B column chromatography indicated a molecular mass of 23 kDa, indicating that the value is similar to those of other PP4 enzyme reported to date. Isoelectric point of PP27 was p15.2. The protein was found to inhibit the coagulation time in a concentration-dependent manner. PP27 was acted as a vascular anticoagulant of annexin type, inhibits the blood clotting process by binding of the essential lipids in a reaction which is dependent on $Ca2^+$ ions. In the presence of $Ca2^+$ ions, PP27 combines with platelet membranes neutralizing their procoagulant effect. Coagulation triggered by the addition of thromboplastin/ lipid- mixtures is extinguished by PP27.