• 미생물학회지
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• 제30권6호
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• pp.501-506
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• 1992

An alkalophilic Vibrio sp. RH530 showing high proteolytic activity was isolated form soil samples by enrichment culture. The activity staining using gelatin SDS- polyacrylamide gel electrophoresis (PAGE ) revealed that the strain produced an alkaline major protease (Apr B) with a size of 27 kDa, and at least six minor proteases. The apparent sizes of four of the minor proteases were approximately 45, 28, 22 and 19 kDa. Apr B and five of the minor proteases were inhibited by serine protease inhibitors including PMSF and DFP, suggesting that they are serine proteases. One of the minor proteases was inhibited by metalloprotease inhibitors, not by serine protease inhibitors, indicating it to be a metalloprotease. Furthermore, the activities of Apr B and Prt 3 were not inhibited by SDS in the reaction mixture. The production of Apr B and some of the minor proteases was specifically affected by culture temperature (30 to 37.deg.C) and pH (7 to 10). The production of Apr B. Prt 2, Prt 5 and Prt 6 was mainly affected by culture temperature, while Prt 4 by culture pH. Prt 1 and Prt 3 were not affected by neither of these factors.

• BMB Reports
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• 제38권5호
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• pp.555-562
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• 2005

Single nucleotide polymorphisms (SNPs) are becoming the most common type of markers used in genetic analysis. In the present report a SNP has been chosen to test the applicability of Real Time PCR to discriminate and quantify SNPs alleles on DNA pools. Amplification Refractory Mutation System (ARMS) and Mismatch Amplification Mutation Assay (MAMA) has been applied. Each assay has been pre-validated testing specificity and performances (linearity, PCR efficiency, interference limit, limit of detection, limit of quantification, precision and accuracy). Both the approaches achieve a precise and accurate estimation of the allele frequencies on pooled DNA samples in the range from 5% to 95% and don't require standard curves or calibrators. The lowest measurement that could be significantly distinguished from the background noise has been determined around the 1% for both the approaches, allowing to extend the range of quantifications from 1% to 99%. Furthermore applicability of Real Time PCR assays for general diagnostic purposes is discussed.

• BMB Reports
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• 제42권6호
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• pp.367-372
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• 2009

In this study, we showed that WAX9D, a nonspecific lipid-transfer protein found in broccoli, binds palmitate (C16) and stearate (C18) with dissociation constants of 0.56 ${\mu}M$ and 0.52 ${\mu}M$, respectively. WAX9D was fused to thioredoxin protein by genetic manipulation to enhance its solubility. The data revealed strong interaction of Trx-WAX9D with palmitate and stearate. The dissociation constants of Trx-WAX9D for palmitate and stearate were 1.1 ${\mu}M$ and 6.4 ${\mu}M$, respectively. The calculated number of binding sites for palmitate and stearate was 2.5 to 2.7, indicating that Trx-WAX9D can bind three molecules of fatty acids. Additionally, Trx-WAX9D was shown to inhibit the apoptotic effect of palmitate in endothelial cells. Our data using Trx-WAX9D provide insight into the broad spectrum of its biological applications with specific palmitate binding.

• BMB Reports
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• 제43권6호
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• pp.419-426
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• 2010

Aeromonas hydrophila is a bacterial pathogen that infects a large number of eukaryotes, including humans. The UDP-galactose 4'-epimerase (GalE) catalyzes interconversion of UDP-galactose to UDP-glucose and plays a key role in lipopolysaccharide biosynthesis. This makes it an important virulence determinant, and therefore a potential drug target. Our earlier studies revealed that unlike other GalEs, GalE of A. hydrophila exists as a monomer. This uniqueness necessitated elucidation of its structure and active site. Chemical modification of the 6xHis-rGalE demonstrated the role of histidine residue in catalysis and that it did not constitute the substrate binding pocket. Loss of the 6xHis-rGalE activity and coenzyme fluorescence with thiol modifying reagents established the role of two distinct vicinal thiols in catalysis. Chemical modification studies revealed arginine to be essential for catalysis. Site-directed mutagenesis indicated Tyr149 and Lys153 to be involved in catalysis. Use of glycerol as a cosolvent enhanced the GalE thermostability significantly.

• Molecular & Cellular Toxicology
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• 제5권1호
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• pp.14-22
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• 2009

In vitro cytotoxicity of 23 alkyl phenols (APs) on human cervical cancer cell lines (HeLa) was determined using the lactate dehydrogenase (LDH) cytotoxicity assay. Two different sets of descriptors were used to construct the calibration model based on Genetic Algorithm-Multiple Linear Regression (GA-MLR) based on the experimental data. A statistically robust Structure-Activity Relationships (QSAR) model was achieved ($R^2$=95.05%, $Q^2_{LOO}$=91.23%, F=72.02 and SE= 0.046) using three Dragon descriptors based on Me (0D-Constitutional descriptor), BELp8 (2D-Burden eigenvalue descriptor) and HATS8p (3D-GETAWAY descriptor). However, external validation could not fully prove its validity of the selected QSAR in characterization of the cytotoxicity of APs towards HeLa cells. Nevertheless, the cytotoxicity profiles showed a finding that 4-n-octylphenol (4-NOP), 4-tert-octyl-phenol (4-TOP), 4-n-nonylphenol (4-NNP) had a more potent cytotoxic effect than other APs tested, inferring that increased length and molecular bulkiness of the substituent had important influence on the LDH cytotoxicity.

• The Plant Pathology Journal
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• 제36권4호
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• pp.364-377
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• 2020

Sugarcane is an important sugar crop contributes more than 80% of world sugar production. Mosaic, leaf fleck, and yellow leaf (YL) are the major viral diseases affecting sugarcane, amongst YL occurrence is widely reported in all the sugarcane growing countries. It is caused by Sugarcane yellow leaf virus (SCYLV) and detailed works were done on complete genome characterization, transmission, and management. However, in countries like Egypt, South Africa, Cuba, Mauritius and Hawaii, the disease was reported to the cause of sugarcane yellow leaf phytoplasma (SCYP) and/or SCYLV as single/combined infections. Hence, we have investigated in detail to identify the exact Candidatus phytoplasma taxon associated in Indian cultivars affected with YL. The sequencing results and the restriction fragment length polymorphism pattern of the PCR products using the universal phytoplasma primers confirmed presence of sugarcane grassy shoot (SCGS) phytoplasma (16SrXI group) in the YL-affected plants. Mixed infection of SCYLV and SCGS phytoplasma was estimated as 32.8% in YL affected plants. Evolutionary genetic relationship between SCYP and SCGS phytoplasma representatively taken from different countries showed that SCYP from South Africa and Cuba were diverged from others and had a highest similarity with SCGS phytoplasma. Although we wanted to identify SCYP from YL affected Indian sugarcane cultivars, the study clearly indicated a clear absence of SCYP in YL affected plants and we found SCYLV as the primary cause for the disease.

• Food Science and Biotechnology
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• 제15권6호
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• pp.877-883
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• 2006

The immunostimulatory activities of Lactobacillus plantarum, the major microorganism in kimchi fermentations were investigated. Five strains of L. plantarum exhibited weak immunopotentiating activity, but L. plantarum PS-21 showed as strong a mitogenic activity as Bifidobacterium adolescentis M101-4, a known positive strain. It is of interest that, L. plantarum PS-21 stimulated proliferation of Peyer's patch cells, one of the most important tissues in the gut-associated lymphoreticular system. Cell' wall fractions from L. plantarum PS-21 also showed strong mitogenic activity compared with the soluble cytoplasmic fraction. A peptidoglycan fraction (PG) extracted from the cell wall of L. plantarum PS-21 was identified as an active mitogenic component when used in murine lymph node and spleen cell test systems. PG showed dose-dependent mitogenic activity and significantly enhanced antibody production in lymph node cells when studied in vitro. The lysosomal enzyme activity of murine peritoneal macrophages was increased when analyzed following injection of PG to the host animal. Furthermore, PG enhanced the production of cytokines such ($TNF-{\alpha}$ and IL-6) in the in vitro culture of RAW 264.7 macrophage cells.

• Asian-Australasian Journal of Animal Sciences
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• 제28권2호
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• pp.158-165
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• 2015

Marbling is an important trait regarding the quality of beef. Analysis of beef cattle transcriptome and its expression profile data are essential to extend the genetic information resources and would support further studies on beef cattle. RNA sequencing was performed in beef cattle using the Illumina High-Seq2000 platform. Approximately 251.58 million clean reads were generated from a high marbling (H) group and low marbling (L) group. Approximately 80.12% of the 19,994 bovine genes (protein coding) were detected in all samples, and 749 genes exhibited differential expression between the H and L groups based on fold change (>1.5-fold, p<0.05). Multiple gene ontology terms and biological pathways were found significantly enriched among the differentially expressed genes. The transcriptome data will facilitate future functional studies on marbling formation in beef cattle and may be applied to improve breeding programs for cattle and closely related mammals.

• Animal cells and systems
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• 제3권4호
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• pp.399-405
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• 1999

Characteristics of the food isolates and the clinical specimens isolates of E. coli harboring virulence factor and their correlations were analyzed. The predominant serogroup were 08 and 027 in the food isolates and 06 and 018 in the clinical isolates, respectively, showing the different patterns in serogrouping between them. In the test of antibiotic susceptibility, the food isolates were resistant to cephalothin, streptomycin, tetracycline and minocycline, and the clinical isolates were resistant to ampicillin, carbenicillin, streptomycin, cephalothin, trimethoprim/sulfamethoxazole, tetracyclino and minocycline, respectively. It shows that E.coli isolated from food sources and clinical specimens might be correlated. Plasmid profile in the food and clinical isolates showed wide diversity. Especially, large sized plasmid DNA such as 60 MDa, 90 MDa and 120 MDa were observed. The plasmid DNA (60 MDa) containing a gene encoding hemolysin was found in 43% of the food isolates and 35% of the clinical isolates. To study chromosomal homology, PFGE analysis was performed, showing different restriction patterns by Xbal. This result indicates that there were no genetic correlations between the foods and the clinical isolates.

• 대한바이러스학회지
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• 제28권2호
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• pp.139-146
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• 1998

Recombinant polioviruses have been developed by many research groups for use as vaccine vector because poliovirus induces mucosal immunity as well as humoral immunity through oral uptake. We assessed the potential use of poliovirus as a T-cell epitope carrier. Recombinant poliovirus V129 5L was constructed to have a substituted T-helper epitope from the core protein of Hepatitis B virus at neutralization antigenic site 1 on its VP1 capsid protein. The recombinant virus replicated less efficiently than type 1 poliovirus Mahoney strain. The V129 5L formed a little smaller plaques than the Mahoney strain and showed some 1.25 log unit lower titer at the peak in the one-step growth kinetics though it had similar growth profile to that of the Mahoney strain. Since V129 5L recombinant virus was genetically stable even after 24 successive passages in HeLa cells, the antigenic site 1 on VP1 capsid protein was confirmed for its ability of carrying T cell epitope. The genetic stability of V129 5L also indicated that recombinant poliovirus can be successfully utilized for the development of the multivalent vaccines.