• Korean Journal of Microbiology
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• v.43 no.3
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• pp.166-172
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• 2007

ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confer the antibiotic resistance on micro-organisms ranging from antibiotic producers to pathogens and are classified into monomethyltransferase and dimethyltransferase. To investigate the differences between mono- and dimethyltransferase, tirD, a representative monomethylase gene was cloned in Escherichia coli from Streptomyces fradiae which contains ermSF, dimethylase gene as well to overexpress the TlrD for the first time. T7 promoter driven expression system successfully overexpress tlrD as a insoluble aggregate at $37^{\circ}C$ accumulating to around 55% of the total cell protein but unlike ErmSF, culturing at temperature as low as $18^{\circ}C$ did not make insoluble aggregate of protein into soluble protein. Coexpression of Thioredoxin and GroESL, chaperone was not helpful in turning into soluble protein either as in case of ErmSF. These results might suggest that differences between mono- and dimethylase could be investigated on the basis of the characteristics of protein structure. However, a very small amount of soluble protein which could not be detected by SDS-PAGE conferred antibiotic resistance on E. coli as in ErmSF which was expected from the activity exerted by monmethylase in a cell.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.165-171
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• 2006

ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA to confer the resistance to MLS (macrolide-lincosamide-streptogramin B) antibiotics on microorganism ranging from antibiotic producers to pathogens. To define the functional role of peptide segment encompassing amino acid residues 1 to 25 in NTER (N-terminal end region) of ErmSF, one of the ERM proteins, DNA fragment encoding mutant protein deprived of that peptide was cloned and overexpressed in E. coli to obtain a purified soluble form protein to the apparent homogeneity in the yield of 12.65 mg per liter of culture. The in vitro activity of mutant protein was found to be 85% compared to wild type ErmSF, suggesting that this peptide interact with substrate to affect the enzyme activity. This diminished activity of mutant protein caused the delayed expression of antibiotic resistance in vivo, that at fIrst cells expressing mutant protein showed the retarded growth due to the antibiotic action but with time cells inhibited by antibiotic gradually recovered the viability to exert the resistance to the same extent as those with wild type protein.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.193-198
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• 2008

ErmSF is one of the proteins which are produced by Streptomyces fradiae to avoid suicide by its autogenous macrolide antibiotic, tylosin and one of ERM proteins which are responsible for transferring the methyl group to $A_{2058}$ (Escherichia coli coordinate) in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confers the antibiotic resistance on microorganisms ranging from antibiotic producers to pathogens. ErmSF contains an extra N-terminal end region (NTER), which is unique to ErmSF and 25% of amino acids of which is arginine known well to interact with RNA. Noticeably, arginine is concentrated in $^{58}RARR^{61}$ and functional role of each arginine in this motif was investigated through deletion and site-directed mutagenesis and the activity of mutant proteins in cell R60 and R61 was found to play an important role in enzyme activity through the study with deletion mutant up to R60 and R61. With the site-directed mutagenesis using deletion mutant of 1 to 59 (R60A, R61A, and RR60, 61AA), R60 was found more important than R61 but R61 was necessary for the proper activity of R60 and vice versa. And these amino acids were presumed to assume a secondary structure of $\alpha$-helix.

• Biomolecules & Therapeutics
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• v.1 no.2
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• pp.177-182
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• 1993

From 84 clinical isolates of Staphylococcus species, ten strains showing inducible resistance to MLS antibiotics were selected by disk agar diffusion method. Colony hybridization was executed using two MLS inducible resistance genes, ermA and ermC, previously identified from S. aureus as probes. S. hemolyticus 401 and S. epidermidis 542 whose genes were not homologous to those probes were finally selected. It was determined that the resistance genes of S. hemolyticus 401 and S. epidermidis 542 were not homologous to ermA, ermC and ermAM by Southern hybridization. S. epidermidis 542 had a plasmid DNA. To know if the plasmid may have genes related to inducible resistance, it was attempted to transform B. subtilis BR151 and S. aureus RN4220 with the plasmid prepared from S. epidermidis 542. It was shown that the gene related to inducible resistance to MLS antibiotics did not exist in this plasmid. These results indicate that two clinical isolates of S. hemolyticus 401 and S. epidermidis 542 had novel genes which were not homologous to MLS resistance genes identified previously. It was assumed that these genes may exist in chromosomal DNA.

• Proceedings of KOSOMES biannual meeting
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• 2017.11a
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• pp.277-277
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• 2017

• YAKHAK HOEJI
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• v.32 no.4
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• pp.213-221
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• 1988

Inducible MLS resistance gene of Bacillus licheniformis specified by erm K was subcloned in Bacillus subtilis and the DNA sequence corresponding to its control region was determined. The determinant erm K was in Pvu II=Hind III fragment, which was 1.3 kb. The leader region is capable of forming a complex series of inverted complementary repeat sequences (ICRS) centering on at least six axes of symmetry, some of them mutually exclusive, in a way that resulted ultimately in post-transcriptional unmasking of the ribosome loading site for methylase synthesis.

• YAKHAK HOEJI
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• v.41 no.4
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• pp.533-537
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• 1997

The characteristics of the resistance phenotypes of Bacillus subtilis having ermk and its terminator region mutants were determined. Wild type ermK(pEC101) and pECMT109(methylase SD-region mutant) showed typical inducible resistance phenotype. pECMF1(terminator1 region mutant) and pECMT2(terminator2 region mutant) showed constitutive resistance to Kitasamycin but inducible resistance to tylosin. In contrast, pECMT3(terminator1 and terminator2 double mutant) and pECMT309(terminator1, terminator2 and methylase SD region triple mutant) showed constitutive resistance both to kitasamycin and tylosin.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.335.1-335.1
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• 2002

The ermK gene from Bacillus lichenformis encodes an inducible rANA methylase that confers resistance to the macrolide-lincosamide-streptograminB antibiotics. The ermKmANA leader sequence has a total length of 357 nucleotides and encodes a 14-amino acid leader peptide together with its ribosome binding site. The secondary structure of erm leader RNA and a leader peptide have been reported as the elements that control expression. (omitted)

• Proceedings of the Korean Institute Of Construction Engineering and Management
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• 2007.11a
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• pp.539-544
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• 2007

Owing to diversification of social desire and increase of demand as economic growth, construction industry recently is trending toward diversification, complication, gigantism. It means that is changing to difficult environment as existing construction company operation. Specially, some big construction companies are promoting get down to construction business risk management skill & development. Enterprise risk management system, recognized to risk portfolio, is suggested. instead of individually risk management. This study indicates ERM basic model, considering construction risk character, for apply to ERM to field of construction . And it is including analysis of recognition level and reality about ERM in construction company.

• Proceedings of the PSK Conference
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• 2000.10a
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• pp.204.1-204.1
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• 2000