• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1377-1383
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• 2006

Among 46 Acinetobacter baumannii isolates collected in 2004, two imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Republic of Korea. Two carbapenemase-producing isolates were further investigated to determine the mechanism of resistance. These isolates were analyzed by antibiotic susceptibility testing, microbiological tests of carbapenemase activity, determination of pI, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. Two cases of infection by A. baumannii producing the IMP-1 ${\beta}$-lactamase were detected. The isolates were characterized by a modified cloverleaf synergy test and EDTA-disk synergy test. Isoelectric focusing of crude bacterial extracts revealed nitrocefin-positive bands with a pI value of 9.0. PCR amplification and characterization of the amplicons by direct sequencing indicated that the isolates carried a $bla_{IMP-l}$ determinant. The isolates were characterized by a multidrug resistance phenotype, including penicillins, extended-spectrum cephalosporins, carbapenems, and aminoglycosides. These results indicate that the observed imipenem resistance of two Korean A. baumannii isolates was due to the spread of an IMP-1-producing clone. Our microbiological test of carbapenemase activity is simple to screen class B metallo-${\beta}$-lactamase-producing clinical isolates to determine their clinical impact and to prevent further spread. This study shows that the $bla_{IMP-l}$ resistance determinant, which is emerging in Korea, may become an emerging therapeutic problem, since clinicians are advised not to use extended-spectrum cephalosporins, imipenem, and aminoglycosides. This observation emphasizes the importance of having effective control measures in Asian hospitals, such as early detection of colonized patients, isolation procedures, and a judicious use of antibiotics.

• Analytical Science and Technology
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• v.16 no.1
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• pp.1-11
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• 2003

This research examined the presence of hazardous materials in chemical workplace field using an integrated chemical/biological monitoring. Chemical workplace field air for volatile organic compounds (VOCs) analysis was collected using a collection tube packed with Tena.x TA adsorbent 400 mg. Workplace field air samples were analyzed by gas chromatography/mass spectrometry (GC/MS). Simultaneously, Tradescantia BNL 4430 clone was exposed in situ to monitor hazardous materials in chemical workplace field. GC/MS analysis showed the presence of various VOCs such as trichloroethylene, toluene, ethylbenzene, (m,p,o)-xylenes, styrene, 1,3,5-trimethylbenzene, and 1,2,4-trimethylbenzene. The results showed that in situ monitoring of VOCs with the Tradescantia-micronucleus (Trad-MCN) assay gave positive results in chemical workplace field and negative response at outdoor air. In conclusion, inhalation of these field air by workers may affect chronic demage to their health by inducing micronuclei formation in Tradescantia pollen mother cells. The combination of chemical/biological monitoring is very effective to evaluate hazardous materials in workplace field and can be alternatively used for screening hazardous materials.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.845-858
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• 1996

The purpose of this study was to establish early diagnostic methods for the detection of Aujeszky's disease viral antigens and nucleic acid in nasal cells, and buffy coats from experimentally infected living pigs by a combination of immunocytochemistry, in situ hybridization with digoxigenin(DIG)-labled probe and electron microscopy. Forty days old piglets were inoculated intranasally with $10^{7.0}TCID_{50}$ of Aujeszky's disease virus (ADV, NYJ-1-87 strain). The viral antigens and nucleic acid of ADV were detected in nasal cells, and buffy coat for 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopical method. The results were compared with conventional methods such as a porcine Aujeszky's disease serodiagnostic(PAD) kit, neutralization test(NT) and virus isolation. 1. The viral antigens, nucleic acids and capsids of ADV were detected in nasal cells, buffy coats from 3 days to 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopy, respectively. 2. When viral antigens were detected by the immunocytochemical technique, a diffuse brown deposit was observed in the nucleus and cytoplasm of nasal cells, buffy coats and PK-15 cells under a microscope. 3. DIG-labeled DNA probe was prepared by amplification of conserved sequence of recombinant ADV-gp50 clone with polymerase chain reacction. When ADV-DNA was detected by ISH with DIG-labeled probe, purplish blue pigmentation were observed in the nuclei and cytoplasms of ADV-infected cells under a microscope. Positive signals were observed in nasal cells and in the buffy coat and PK-15 cells at the first day after inoculation. 4. Where ADV-capsids were detected by transmission electron microscopical method, aggregation of capsids was observed in the nuclei and cytoplasms of nasal cells, buffy coats and PK-15 cells. The results suggested that these methods were considered as the highly sensitive and reliable tools for rapid and confirmative diagnosis of Aujeszky's disease in living pigs.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.420-427
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• 2018

The orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase (Sc YRS) and a variant of E. coli initiator tRNA, fMam $tRNA_{CUA}$ which recognizes the amber stop codon is an effective tool for site-specific incorporation of unnatural amino acids into the protein in E. coli. To evolve the amber suppression activity of the orthogonal pair, we generated a mutant library of Sc YRS by randomizing two amino acids at 320 and 321 which involve recognition of the first base of anticodon in fMam $tRNA_{CUA}$. Two positive clones are selected from the library screening with chloramphenicol resistance mediated by amber suppression. They showed growth resistance against high concentration of chloramphenicol and their $IC_{50}$ values were approximately 1.7~2.3 fold higher than the wild type YRS. In vivo amber suppression assay reveals that mutant YRS-3 (mYRS-3) clone containing amino acid substitutions of P320A and D321A showed 6.5-fold higher activity of amber suppression compared with the wild type. In addition, in vitro aminoacylation kinetics of mYRS-3 also showed approximately 7-fold higher activity than the wild type, and the enhancement was mainly due to the increase of tRNA binding affinity. These results demonstrate that optimization of anticodon recognition by engineered aminoacyl tRNA synthetase improves the efficiency of unnatural amino acid incorporation in response to nonsense codon.

• Journal of Plant Biotechnology
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• v.37 no.3
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• pp.313-318
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• 2010

To develop edible vaccines for swine, the embryogenic calli (type II) derived from HiII genotype were inoculated with A. tumefaciens strain C58C1 containing the binary vector pMYV611, 613, 616, and V621, 622 and 623 respectively. Six of those vectors carry nptII gene which confers resistance to paromomycin and apxIIA gene producing ApxII toxin which is generated in various serum types of A. pleuropneumoniae as a target gene. The 4,120 callus clones for pMYV611, 5,959 callus clones for pMYV613, 7,581 callus clones for pMYV616, 52,329 callus clones for V621, 48,948 callus clones for V622, and 56,188 callus clones for V623 were inoculated. The frequency of positive response clone was confirmed into range of 2.3% - 4.4% for each vectors by NPTII ELISA kit assay, and the selected callus clones of them were finally 3 callus clones from pMYV611 (0.07%), 4 callus clones from pMYV613 (0.07%), 2 callus clones from pMYV616 (0.03%), 51 callus clones from V621 (0.1%), 72 callus clones from V622 (0.15%), and 102 callus clones from V623 (0.18%) respectively. From the selected callus clones of each binary vector, the integration of the apxIIA gene into maize genome was detected from 2 plants of pMYV613 and 2 plants of V623 by Southern blot analysis.

• Journal of Life Science
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• v.18 no.11
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• pp.1592-1599
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• 2008

The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

• Asian Journal of Turfgrass Science
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• v.1 no.1
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• pp.7-17
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• 1987

The experiment with two levels of nitrogen (0. and 300kg / ha / year) and two levels of clipping height (1.5cm and 4cm) was conducted on the field during the period 3 June to 23 October 1985. Clonal lines of korean lawngrass ( Zoysia japonica Steud.) and manilagrass ( Zoysia matrella ( L.) Merr.)of Daejon origin were established in June, as individual clone in rows 30cm apart with a 40cm spacing between clones, actually 4 clones each plot. The results obtained were as follows : 1. When no nitrogen was applied to korean lawngrass, leaf blade which appeared during the August / early September period remained green for a period of about 10 weeks and even leaves emerged in late September lived for 42 days. However, leaf longevity did not exceed 8 weeks as nitrogen was applied. In contrast the leaf longevity of manilagrass which emerged during the mid - August / early September period was 11 weeks and, under the nitrogen applied, 9 weeks, indicating that the life - saen of individual leaf of manilagrass may be longer than that of korean lawngrass. Meanwhile, clipping height had no effect on the leaf longevity in both grasses. 2. During the July / August period, tiller number, green leaf number and DM weight of korean lawngrass were increased significantly with fertilizer nitrogen, but were not with two levels of clipping height. This trend was reversed after late September : no effect of nitrogen was appeared. Instead, lax clipping increased tiller number, green leaf number and DM weight. Green leaves stimulated by lax clipping resulted in the occurrance of more dead leaves in late October. 3. The increase of tiller number, green leaf number, and DM weight of korean lawngrass due to nitrogen application appeared to be of significance in early September. Unlike korean lawngrass, however, this significant increase was maintained to late October when new green leaves still emerge. Clipping height had little effect on the growth of manilagrass by early September, but since then, lax clipping stimulated leaf appearance, possibly resulting in a remained green color of manilagrass turf. 4. Among the stolons outgrown until early September, the primary stolon was not influenced by nitrogen and clipping treatments to produce only 2 - 3 stolons. However, 1st branch stolon as affected by nitrogen increased significantly, so most of stolons which occurred consisted of 1st branch stolon. 5. Until early September, stolon length obtained at nil nitrogen level was chiefly caused by lengthening the primary stolons. By applying nitrogen the primary stolons of korean lawngrass was longer than 1st branch stolons when severe clipping was involved and in turn, shorter than 1st branch stolons when lax clipping was concerned. In manilagrass, 1st branch stolons were much longer than the primary stolons when turf was clipped severely but in conditions of lax clipping, there was little difference in length between primary and 1st branch stolons. 6. Stolon nodes of both korean lawngrass and manilagrass were positively influenced by nitrogen, but no particular increases by imposing clipping height treatment was marked in manilagrass. Although the stolon of korean lawngrass was grown until late october, the growth stimulated by nitrogen was not so remarkable as to exceed that a by nil N. 7. The thickness of korean lawngrass and manilagrass was greatest in late September, but that of manilagrass did not differ significantly from that in late October. 8. The response of stolon length of korean lawngrass to lax clippings was not so great in late October as to that to severe clippings. On the other hand, the positive effect of lax clippings to stolon length in m anilagrass was confirmed even in late October.