• Korean Journal of Microbiology
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• v.28 no.3
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• pp.181-187
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• 1990

STA gene coding glucoamylase was introduced into haploid Saccharomyces cerevisiae SHY3 and polyploid Saccharomyces cerevisiae 54. We constructed the recombinant plasmid by substituting the promoter region of alcohol dehydrogenase isoenzyme I gene for that of STA gene to increase the expression of STA gene and found that the activity of glucoamylase was increased in transformants. The plasmid stability was improved remarkably when we got the STA gene into the plasmid which had centromere. The activity of glucoamylase and transformation frequency of it, however, was decreased because of low copy number. Industrial polyploid strain was transformed with the recombinant plasmid having the $2\mu$ origin of replication and STA gene. It produced more alcohol than host when fermented in liquefied starch media. The industrial strain, however, was not transformed with the autonomously replicating plasmid containing centromere.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.48-53
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• 1993

For the purpose to improve the glucoamylase productivity of Saccharomyces diastaticus, we integrated STA 1 gene into chromosomal DNA of S. diastaticus using YIp vector. After construction of Ylp-STA by the subcloning of STAI (5.3 kb) into YIp5 vector, S. diastaticus GMT-II(a. ura3. STAJ) was transformed by Ylp-STA through homologous recombination at the chromosomal STAJ gene. So we obtained the tram formants that glucoamylase productivity was increased maximum six fold. These strains transformed by the multi-copy integration of Ylp-STA in chromosomal DNA were confirmed by Southern hybridization. And the integrated Ylp-STA was maintained stably during 30 mitotic divisions.

• Korean Journal of Microbiology
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• v.30 no.5
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• pp.403-409
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• 1992

For the development of a glucanolytic yeast strain. the seceretion of endo-1.4-$\beta$-D-glucanase (CMCase) of Bacillus subtilis was performed in yeast using glucoamylase gene (STA1) of Saccharomyces diastaticus. A 1.7 kb-DNA fragment of STA1 gene containing authentic promoter, signal sequence, threonine serine-rich (TS) region and N-terminal region (98 amino acids) of mature glucoamylase was ligated to YEp 24. E. coli-yeast shuttle vector. And then. CMCase structural gene of B. subtilis was fused in frame with the 1.7 kb-DNA fragment of STA1 gene, resulting in recombinant plasmid pYES('24. Yeast transformant harboring pYESC24 had no CMCase activity. So. we deleted TS region and N-terminal region of mature glucoamylase existing between signal sequence and CMCase structural gene in pYESC24. consequently constructed recombinant plasmid pYESC11. The yeast transformed with the newly constructed recombinant plasmid pYESC11 efficiently secreted CMCase to extracellular medium. After 4 days culture. total CMCase activity of this transformant was 44.7 units/ml and over 93% of total CMCase activity was detected in culture supernatant.

• Proceedings of the Korean Society of Crop Science Conference
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• 1997.05a
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• pp.26-27
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• 1997

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.606-610
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• 1989

For the purpose of improving glucoamylase productivity of Saccharomyces diastaticus, useful yeast in direct ethanol fermentation of starch, the effects of growth rate on the plasmid stability and glucoamylase productivity of S. diastaticus harboring recombinant plasmid pYES 18 containing glucoamylase gene STA 1 were investigated. In a selective medium, the recombinant plasmids were maintained stably at constant level but glucoamylase productivity was very low. On the other hand, in the complex medium containing starch, growth rate of the cell was stimulated by the supplementation of glucose and plasmid stability was improved by growth stimulation. We can conclude that glucoamylase productivity of S. diastaticus harboring the recombinant plasmid was increased as the maintaining of high plasmid stability in the cell.

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.301-310
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• 2000

The antimicrobial susceptibility, genotypes of enterotoxins(LT, STa) and pili(K88, 987P), and plasmid profiles were investigated with 102 Escherichia coli isolated from piglets showing diarrhea in Korea. Almost of them were susceptible to ceftiofur(99%), cefquinone(97.1%). However they showed resistance to bacitracin(100%), streptomycin(98%), vancomycin(97%), trimethoprim/sulfamethoxazole(87.2%), tetracycline(84.3%) in antimicrobial susceptibility test. Moreover, all of the isolates demonstrated resistance to more than 2, and 78% of them were resistant to more than 8 of total 17 drugs. Multiplex PCRs for genotyping of enterotoxins(LT, STa) and pili(K88, 987P) were established with primers designed based on sequences from Genebank. Seventeen strains(16.6%) of the isolates had STa gene, 11 strains(10.8%) of them had both STa and LT genes, and 18 strains(17.8%) had K88 gene. But none of the isolates harbored a gene exclusively encoding LT. The gene encoding 987P pili was not found in all isolates. Fifty-four strains of 102 isolates(52.9%) had plasmid with various sizes ranging from 125kb to 1.1kb. Numbers of plasmid per isolates were also various, from 1 to 9. Distinctive relationship between plasmid profiles and genotypes of enterotoxins and pili in the isolates was not found. These results might provide the basic knowledge to establish strategies for the treatment and prevention of colibacillosis in piglets.

• Journal of Life Science
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• v.7 no.4
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• pp.347-354
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• 1997

Some attempts were made to investigate the gentic characteristics of the brown planthopper(BPH), Nilaparvata lugens Sta${\aa}$l, biotypes. In terms of egg and nymphal periods, egg hatcgability, emergence ratio, and population change of BPH and plant damage of rice cultivars by the infestation of BPH biotypes and their hybrid progenies in field conditions. The egg and nymphal periods of the three of BPH biotypes and their hybrid progenies on Dongjinbyeo with no resistance gene were shorter than on rice cultivars with resistance gene. But the periods of biotype-2(♀) X 2(♂) (E) on Samgangbyeo with Bph-1 gene and biotype-3(♀) X 3(♂)(I) on Mirying 63 with bph-2 gene were as short as on Dongjinbyeo. The egg hatchability and emergence ratio of the three Bph biotypes and their hybrid progenies on Dongjinbyeo were higher than on the other rice cultivars, and those of E on Samgangbyeo and I on Miryang 63 were as high as on Dongjinbyeo. In rice paddy field, biotype-1(♀) X 1(♂)(A)$\sim$I infested on Dongjinbyeo, E onSamgangbyeo, and I on Miryang 63 were increased their population densities remarkably in next generation. Dongjinbyeo infested by A$\sim$I, and Myryang 63 infested by I were hopperburned at 60 days after infestation and had a damage rating of 0.9. But Samgangbyeo infested by E was no hopperburn(damage rating: 7.3) though it showed high BPH population at 60 days after infestation.

• Journal of Life Science
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• v.20 no.5
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• pp.683-690
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• 2010

The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase I, II or III, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and $45^{\circ}C$, respectively, whereas those for the extracellular glucoamylase were 5.0 and $50^{\circ}C$. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.4
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• pp.595-601
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• 2010

This study was conducted to detect and identify Staphylococcus aureus, Vibrio parahaemolyticus, and Salmonella enterica subsp. using simultaneous multiplex polymerase chain reaction (multiplex PCR) assay. 23S rRNA partial gene (S. aureus), tox R gene (V. parahaemolyticus), and inv A gene (S. enterica subsp.) as diagnostic marker gene were suggested, and their amplicon sizes were 482 bp, 368 bp, and 284 bp, respectively. Non specific amplicons by STA-5F/STA-5R primer, ToxR-F/ToxR-R primer, and 139/141 primer were not observed in genomic DNA of pathogen bacteria as Bacillus cereus, Listeria monocytogenes, Escherichia coli, Proteus vulgaris, Streptococcus pyogenes, Candida albicans, and Shigella sonnei. The extracted crude DNA of targeted bacteria was detected as PCR template successfully. The detection limits were $10^5\sim10^4$ CFU/mL and 10 pg of purified genomic DNA of S. aureus, V. parahaemolyticus, and S. enterica subsp. by using simultaneous multiplex PCR.

• Korean Journal of Veterinary Service
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• v.35 no.2
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• pp.99-104
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• 2012

Otitis externa (OE) is a frequent disease in the ear canals of dogs. To identify the pathogens causing OE in dogs and to determine their antimicrobial resistances, specimens were collected from animal hospitals in Daejeon. The isolates were examined by morphological and biochemical tests, 16S rRNA analysis and antimicrobial susceptibility tests. We analyzed correlation between the isolated pathogens and external factors of dogs such as breed, age, gender, ear mite, hair in ears and experience with antibiotic therapy. Thirty three strains of bacteria were isolated from 26 of the 68 heads of dogs with OE. The most isolated bacteria were Enterococcus faecalis (E. faecalis) followed by Staphylococcus aureus (Sta. aureus), Sta. pseudointermedius, E. faecium, E. avium and Streptococcus canis (Strep. canis) in order of frequency of occurrence. Isolation frequency of Enterococcus spp. and Staphylococcus spp. were 51.5% and 45.5%, respectively. E. faecalis and E. faecium isolates showed VanB phenotype, which is resistant to vancomycin but sensitive to teicoplanin were 58% and 25%, respectively. Nine isolates among total twelve isolates of E. faecalis were isolated from the dogs treated with antibiotics. There was no methicillin-resistant Sta. aureus (MRSA), but were MR-Sta. pseudointermedius (MRSP) (57.1%) and vancomycin-resistant (VR)-Sta. pseudointermedius (14.3%) (VRSP) showing VanB phenotype. However, vanA, vanB and vanC genes were not detected in VR isolates from the dogs. Taken together, VR-Enterococcus spp. (VRE) is one of the major pathogens in domestic animals, as well as community-and hospital-acquired infection.