• Title/Summary/Keyword: ErmB

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Expression Profiles of Streptomyces Doxorubicin Biosynthetic Gene Cluster Using DNA Microarray System (DNA Microarray 시스템을 이용한 방선균 독소루비신 생합성 유전자군의 발현패턴 분석)

  • Kang Seung-Hoon;Kim Myung-Gun;Park Hyun-Joo;Kim Eung-Soo
    • KSBB Journal
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    • v.20 no.3
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    • pp.220-227
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    • 2005
  • Doxorubicin is an anthracycline-family polyketide compound with a very potent anti-cancer activity, typically produced by Streptomyces peucetius. To understand the potential target biosynthetic genes critical for the doxorubicin everproduction, a doxorubicin-specific DNA microarray chip was fabricated and applied to reveal the growth-phase-dependent expression profiles of biosynthetic genes from two doxorubicin-overproducing strains along with the wild-type strain. Two doxorubicin-overproducing 5. peucetius strains were generated via over-expression of a dnrl (a doxorubicin-specific positive regulatory gene) and a doxA (a gene involved in the conversion from daunorubicin to doxorubicin) using a streptomycetes high expression vector containing a strong ermE promoter. Each doxorubicin-overproducing strain was quantitatively compared with the wild-type doxorubicin producer based on the growth-phase-dependent doxorubicin productivity as well as doxorubicin biosynthetic gene expression profiles. The doxorubicin-specific DNA microarray chip data revealed the early-and-steady expressions of the doxorubicin-specific regulatory gene (dnrl), the doxorubicin resistance genes (drrA, drrB, drrC), and the doxorubicin deoxysugar biosynthetic gene (dnmL) are critical for the doxorubicin overproduction in S. peucetius. These results provide that the relationship between the growth-phase-dependent doxorubicin productivity and the doxorubicin biosynthetic gene expression profiles should lead us a rational design of molecular genetic strain improvement strategy.

Analysis of Genotype and Phenotype of Erythromycin Resistance in Enterococci spp. Isolated from Raw Milk Samples (원유시료에서 분리한 장구균의 에리스로마이신 내성 유전자형 및 표현형 분석)

  • Lee, Hye-In;Jung, Jae-Hyuk;Lee, Sang-Jin;Choi, Sung-Sook
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.148-151
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    • 2010
  • The aim of this study was to investigate the erythromycin resistance patterns of Enterococci sp. present in cow milk. A total 110 erythromycin resistant Enterococci were isolated from milk samples; E. faecalis (n=101), E. avium (n=7), and E. durans (n=2). The minimum inhibitory concentration of erythromycin against 110 Enterococci were determined. The 66.3% of Entercocci (n=73) showed high level resistance (${\geq}64$ mg/ml). Among 110 isolates, 86.3% (n=95) showed $cMLS_B$ phenotype and 13.6% (n=15) showed $iMLS_B$ The aim of this study was to investigate the erythromycin resistance patterns of Enterococci sp. present in cow milk. A total 110 erythromycin resistant Enterococci were isolated from milk samples; E. faecalis (n=101), E. avium (n=7), and E. durans (n=2). The minimum inhibitory concentration of erythromycin against 110 Enterococci were determined. The 66.3% of Entercocci (n=73) showed high level resistance (${\geq}64$ mg/ml). Among 110 isolates, 86.3% (n=95) showed $cMLS_B$ phenotype and 13.6% (n=15) showed $iMLS_B$ phenotype. All of isolates have erm(B) determinant, 75.45% (n=83) have mef(A) an efflux system determinant. The majority of Enetrcococci isolated from raw milk samples in northern area of Gyeonggi-Do showed high level of resistance to erythromycin.

Development of a Recombinant Streptomyces griseus with sprA and sprB Genes for Proteolytic Enzyme Production (Streptomyces griseus IFO13350 유래 sprA 및 sprB 유전자를 이용한 Pretense 생산균주 개발)

  • Hwang Ji-Hwan;Lee Chang-Kwon;Lee Kang-Mu;Jo Byoung-Kee;Park Hae-Ryong;Hwang Yong-Il
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.87-92
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    • 2005
  • Pronase, a protease produced for commercial purpose by Streptomyces griseus, was composed of serine protease, alkaline protease, aminopeptidase and carboxypeptidase complex, and it has been widely used as anti-inflammatory drugs for human therapy. In this study, we developed a new integration vector, pHJ101 derived from pSET152, containing strong promoter, ermE, to overexpress a certain protease gene. Specific PCR primers for cloning of sprA (a gene for S. griseus protease A) and sprB (a gene for S. griseus protease B) genes were designed from the basis of nucleotide sequence in databases and amplified by PCR. Plasmid pHJ201 and pHJ202 were constructed by inserting of amplified each gene in a vector pHJ101. S. griseus HA and S. griseus HB were respectively obtained by conjugal process of a parent strain, S. griseus IFO 13350 with the recombinant Escherichia coli harboring plasmid pHJ201 or pHJ202. When protease activity was measured in flask cultivation, produced protease levels of S. griseus HA and S. griseus HB increased about 5.3 times and 5 times, respectively, more than that of parent strain. And, the constructed integrating plasmid pHJ101 was applicable for overexpression of a certain gene in Streptomyces sp.

Distribution of resistance genes against lincomycin of pathogenic bacteria isolated from cultured olive flounder (Paralichthys olivaceus) (양식 넙치에서 분리한 어병세균의 lincomycin에 대한 내성 유전자의 분포)

  • Kim, Ye Ji;Jun, Lyu Jin;Lee, Young Juhn;Ko, Ye Jin;Han, So Ri;Kim, Sung Hyun;Jeong, Joon Bum
    • Journal of fish pathology
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    • v.35 no.1
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    • pp.47-56
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    • 2022
  • Lincomycin as one of the lincosamides antibiotics have been mainly used in human and livestock fields, but have not been used in aquaculture. In this study, the distribution of minimum inhibitory concentration (MIC) values against lincomycin and the detection of the macrolide-lincosamide-streptogramin (MLS) resistance gene were confirmed in bacterial pathogens isolated from cultured olive flounder (Paralichthys olivaceus). Of the 107 strains isolated from Jeju, 36 strains of Gram-positive bacteria and 71 strains of Gram-negative bacteria were identified. Most of Streptococcus spp. was found to have a MIC value of less than or equal to 0.5 ㎍/mL, and Edwardsiella piscicida was found to have a MIC value higher than 1,024 ㎍/mL. V. harveyi and V. alginolyticus mostly showed MIC values of 256 ㎍/mL, but V. scophthalmi displayed values of 8~64 ㎍/mL. In the detection of MLS resistance gene, erm(B) was detected in 9 strains of Streptococcus spp., and erm(A) was confirmed in one strain.

Metabolic Engineering of Rational Screened Saccharopolyspora spinosa for the Enhancement of Spinosyns A and D Production

  • Jha, Amit Kumar;Pokhrel, Anaya Raj;Chaudhary, Amit Kumar;Park, Seong-Whan;Cho, Wan Je;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.37 no.10
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    • pp.727-733
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    • 2014
  • Spinosyns A and D are potent ingredient for insect control with exceptional safety to non-target organisms. It consists of a 21-carbon tetracyclic lactone with forosamine and tri-Omethylated rhamnose which are derived from S-adenosyl-methionine. Although previous studies have revealed the involvement of metK1 (S-adenosylmethionine synthetase), rmbA (glucose-1-phosphate thymidylyltransferase), and rmbB (TDP-D-glucose-4, 6-dehydratase) in the biosynthesis of spinosad, expression of these genes into rational screened Saccharopolyspora spinosa (S. spinosa MUV) has not been elucidated till date. In the present study, S. spinosa MUV was developed to utilize for metabolic engineering. The yield of spinosyns A and D in S. spinosa MUV was $244mgL^{-1}$ and $129mgL^{-1}$, which was 4.88-fold and 4.77-fold higher than that in the wild-type ($50mgL^{-1}$ and $27mgL^{-1}$), respectively. To achieve the better production; positive regulator metK1-sp, rmbA and rmbB genes from Streptomyces peucetius, were expressed and co-expressed in S. spinosa MUV under the control of strong $ermE^*$ promoter, using an integration vector pSET152 and expression vector pIBR25, respectively. Here-with, the genetically engineered strain of S. spinosa MUV, produce spinosyns A and D up to $372/217mgL^{-1}$ that is 7.44/8.03-fold greater than that of wild type. This result demonstrates the use of metabolic engineering on rationally developed high producing natural variants for the production.

Characterization of S. aureus Showing MLS Resistance with Foggy D Shape (fMLS) (안개 D형 저지원을 갖는 MLS 내성 황색 포도상 구균의 분포와 내성 기전)

  • Yoon, Eun-Jeong;Kim, Hyun-Jee;Kwon, Ae-Ran;Choi, Sung-Sook;Shim, Mi-Ja;Choi, Eung-Chil
    • YAKHAK HOEJI
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    • v.50 no.3
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    • pp.199-203
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    • 2006
  • A new type of macrolides, lincosamides and streptogramin B antibiotics (MLS) resistance, showing the characteristic phenotype growing within the inhibition zone area around the clindamycin disk, was identified among clinically isolated Staphylococcus aureus. We named the phenotype as MLS resistance with foggy-D shape (fMLS). The average frequency of MLS isolates was 9%. All of fMLS isolates have only erm(A) for the resistance determinant. The growth pattern of the challenged MLS isolate looks intermediate phase between the patterns of inducible MLS resistance and constitutive MLS resistance.

Emergence of macrolide resistance and clinical use of macrolide antimicrobials in children (Macrolide계 항균제 내성 출현과 소아에서의 임상적 적용)

  • Choi, Eun Hwa
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1031-1037
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    • 2008
  • Macrolide antimicrobial agents including erythromycin, roxithromycin, clarithromycin, and azithromycin are commonly used in the treatment of respiratory tract infections in children. Newer macrolides that have structural modifications of older drug erythromycin show improved change in the spectrum of activity, dosing, and administration. However, recent studies reported that increasing use of macrolide antibiotics is the main force driving the development of macrolide resistance in streptococci. In particular, azithromycin use is more likely to select for macrolide resistance with Streptococcus pneumoniae than is clarithromycin use, a possible reflection of its much longer half life. Recently, erythromycin resistance rates of S. pneumoniae and Streptococcus pyogenes are rapidly increasing in Korea. Two main mechanisms of acquired macrolide resistance have been described, altered binding site on the bacterial ribosome encoded by the ermB gene and active macrolide efflux pump encoded by the mef gene. Relationship between the susceptibility of S. pneumoniae and the response to macrolides has been shown in studies of acute otitis media, but less clear in cases of pneumonia. This article reviews the spectrum of activity, pharmacokinetic properties, mechanisms of action and resistance, and clinical implication of resistance on the treatment of respiratory tract infections in children.

The First Korean Case of Nontuberculous Mycobacterial Lung Disease Caused by Mycobacterium abscessus Subspecies bolletii in a Patient with Bronchiectasis

  • Jeong, Byeong-Ho;Kim, Su-Young;Jeon, Kyeongman;Huh, Hee Jae;Ki, Chang-Seok;Lee, Nam Yong;Shin, Sung Jae;Koh, Won-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.76 no.1
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    • pp.30-33
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    • 2014
  • We report the first Korean case of lung diseases caused by Mycobacterium abscessus subsp. bolletii in a previously healthy male, except for a previous history of pulmonary tuberculosis and bronchiectasis. All serial isolates are identified as M. abscessus subsp. bolletii by multi-locus sequence analysis based on the hsp65, rpoB, and 16S rRNA fragments. At the genetic level, the isolate has the erm(41) gene with a T28 sequevar, associated with clarithromycin resistance, and no rrl mutation. The isolate is resistant to clarithromycin. Although the symptoms and radiographic findings have improved after combination of antibiotics, the follow-up sputum cultures are persistently positive.

$\"{U}ber$ die kommunikative Funktion von Blickverhalten (시선의 의사소통 기능)

  • Kim Kab-Nyun
    • Koreanishche Zeitschrift fur Deutsche Sprachwissenschaft
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    • v.3
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    • pp.57-74
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    • 2001
  • Kommunikation macht 'gemeinschaftlich'. Sie erscheint v.a. als verbale, d.h. als lautliche und davon abgeleitete Sprache. Lautliche Sprache wird artikulatorisch produziert, akustisch transportiert und auditiv wahrgenommen. Was auch immer ein Mensch einem anderen mitteilen mag, wird ein geringer Teil seiner Botschaft durch Sprache kommuniziert. Neben den akustisch vermittelten Signalen $k\"{o}nnen$ auch nicht-vokale, visuell wahmehmbare Verhaltensweisen eines Interaktanden $f\"{u}r$ einen Beobachter Kommunikationswert annehmen. Der $Kommuni-kationsproze{\ss}$ wird nicht nur durch Senden und Empfangen von akustischen Signalen konstituiert, sondern auch auf der optischen Ebene der gegenseitigen Wahmehmung etabliert. Deshalb sagt man, es sei unmoglich, nicht zu kommunizieren. Denn nicht nur Worte sind das Material jeglicher Kommunikation, sondern auch alle paralinguistischen Phanomene (wie z.B. Tonfall, Schnelligkeit oder Langsarnkeit der Sprache, Pausen, Lachen und Seufzen usw.), $K\"{o}rferhaltung$ usw. In der vorligenden Arbeit bin ich darauf eingegangen, wie die Blickrichtungen einer Person von einem $Gegen\"{u}ber$ sowie einem unbeteiligten Beobachter beurteilt werden. Dabei habe ich versucht zu $erl\"{a}utern$, welche Ursache und Bedeutung das Blickverhalten in der sozialen Interaktion hat. Und mit der Frage, wie dieses Verhalten wahrzunehmen und zu interpretieren ist, habe ich mich $besch\"{a}ftigt$. Die indikative Funtion des Blickverhaltens durch Zu- oder Abwenden des Blickes wurde dabei festgestellt. 'Die Sprache der Augen', 'die Kommunikation mit Augen' scheint eine Vielzahl von Mitteilungen an den sozialen Partnern zu $erm\"{o}glichen$. Die Bedeutung der einzelnen Blicken sind interkulturell verschieden, die Blickverhalten sind teils angeboren, oder sie $m\"{u}ssen$ gelemt werden.

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Characterisation of Phenotypic and Genotypic Antibiotic Resistance Profile of Enterococci from Cheeses in Turkey

  • Kurekci, Cemil;Onen, Sevda Pehlivanlar;Yipel, Mustafa;Aslantas, Ozkan;Gundogdu, Aycan
    • Food Science of Animal Resources
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    • v.36 no.3
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    • pp.352-358
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    • 2016
  • The aim of this study was to determine the prevalence of enterococci in cheese samples and to characterize their antimicrobial resistance profiles as well as the associated resistance genes. A total of 139 enterococci were isolated from 99 cheese samples, the isolates were identified as E. faecalis (61.2%), E. faecium (15.1%), E. gallinarum (12.9%), E. durans (5.0%), E. casseliflavis (2.9%) and E. avium (2.9%). The most frequent antimicrobial resistance observed in enterococci isolates was to lincomycin (88.5%), followed by kanamycin (84.2%), gentamycin (low level, 51.1%), rifampin (46.8%) and tetracycline (33.8%). Among the isolates, the frequencies of high level gentamycin and streptomycin resistant enterococci strains were 2.2% and 5.8%, respectively. Apart from the mentioned antibiotics, low levels of resistance to ciprofloxacin, erythromycin and chloramphenicol were found. Moreover no resistance was observed against penicillin and ampicillin. The antimicrobial resistance genes including tetM, tetL, ermB, cat, aph(3’)-IIIa, ant(6)-Ia and aac(6’)-Ieaph(2”)-Ia were found in enterococci from Turkish cheese samples. In the current study, we provided data for antibiotic resistance and the occurrence of resistance genes among enterococci. Regulatory and quality control programs for milk and other dairy products from farms to retail outlets has to be established and strengthened to monitor trends in antimicrobial resistance among emerging food borne pathogens in Turkey.