• 미생물학회지
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• 제23권4호
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• pp.282-290
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• 1985

Small size antibiotic resistance plasmids having molecular weights less than 10 Mdal were isolated and characterized from ten clinically isolated multiple resistant Staphylococcus aureus. Agarose gel electrophoresis profiles and antibiotic resistance patterns divided these strains into four groups. Strain 2-23-6, the representative strain of a group of five strains conferred two plasmids of molecular weights $1.6{\times}10^6\;dal\;and\;2.0{\times}10^6$ dal. The small plasmid (pSBK 112) specified macrolides, lincosamides and streptogramin type B (MLS) resistance gene which are expressed constitutively. Lage plasmid (pSBK 125) specified chloramphenicol resistance gene which is inducible. Strain 10-5 conferred a $3.0{\times}10^6$ dal plasmid (pSBK 141) which carry an inducible ampicillin resistance gene and strain P-H-2 conferred and $1.6{\times}10^6$ dal plasmid (pSBK 190) which carry a constitutive MLS resistance gene. Strain D-H-1 conferred four plasmids of molecular weights $0.8{\times}10^6$ dal (pSBK 201), $1.6{\times}10^6$ dal (pSBK 202), $2.5{\times}10^6$ dal (pSBK 203), and $1.2{\times}10^7$ dal (pDBK 204), respectively. Among those four plasmids, only pSBK 203 specified chloramphenicol resistance gene. Curing of constitutive MLS resistance using acriding orange or ethidium bromide in 2-23-6 and P-H-2 strains produced 'inducible' MLS resistance strains which are less resistant to MLS than the wild type strains, suggesting that there are two resistance genes in both strains; one is constitutive and the other is inducible.

• 미생물학회지
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• 제24권4호
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• pp.341-351
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• 1986

항생제에 대하여 다중 저항성을 갖는 Shaphylococcus aµreus D-H-1으로부터 chloram phenicol(Cm) 저항성 유전자를 가진것으로 확인된 R-plasmid(pSBK203, 2.5MdaJ.)를 분리하여 Bacillus subtilis BD 170 내에셔 발현시켰으며 이 Cm 저항성 유전자를 cloning하기 위하여 pSBK203상의 제한효소 인식부위를 결정하였다. pSBK203 상의 TaqI 부분 절편 0.3 kb을 pBD9 CZaI 인식부위내에 삽입하여 얻은 재조합 plasmid(pTQ16)와 TaqI부 분절편과 O.lkb 엇갈럼이 있는 RsaI 단일절떤(1. 3 kb} pBR322의 Seal 인식부위에 삽입하여 얻은 재조합 p plasmid(pHW20)에서 Cm 저항성이 획득되었다. pBD9 및 pBR322 상에 삽입된 두 절펀내에 Hinf, Taq I 빛 BglII의 제한효소 인식부위가 존재하였으며 이들 중 행III 인식부위에 의하여 Cm 저항성이 불활성 되었다.

• Clinical and Experimental Pediatrics
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• 제46권1호
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• pp.24-32
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• 2003

목 적 : 황색포도구균(S. aureus)은 건강인의 비강이나 피부에 정상 세균총으로 존재하지만, 병원 내 감염과 항생제 내성이 문제가 되고 있다. 본 연구는 신생아의 비강에서 S. aureus을 분리하여 항생제 감수성의 양상과 내성 관련 유전자를 검출하여 내성의 정도와 내성 관련 유전자의 상관관계를 규명해 보고자 하였으며, 병원성 인자의 검출을 실시하고, arbitrarily-primed polymerase chain reaction(AP-PCR)법에 의한 유전자형별을 통하여 역학적 해석을 시도하였다. 방 법 : 2001년 2월에서 5월 사이에 부산대학교병원 신생아 중환자실 및 신생아실에 입원한 28명을 대상으로 이들의 비강에서 총 120회의 nasal swab을 실시하여 51주의 S. aureus를 분리하였다. 분리균에 대한 항생제 감수성 검사, coagulase 검사과형별, plasmid DNA의 양상, mec 관련 유전자의 분포, 장독소의 유전자 및 TSST-1 독소의 유전자 보유 상태, AP-PCR법으로 유전자 형별을 조사하였다. 결 과 : 1) Methicillin 내성이 23주, vancomycin 내성이 6주였고, 3가지 이상의 약제에 대한 내성이 절반 이상을 차지하였다. 2) 42주가 coagulase 양성이었고, 이중 mecA 및 femA 유전 자를 보유한 12주를 대상으로 한 형별 검사에서 8종(coagulase I-VIII형)으로 분류되었다. 3) mecA 유전자가 22주에서, mecR1 유전자가 18주, mecI 유전자가 1주, femA 유전자가 17주에서 발견되었으며, 이들을 4가지의 유전자형으로 분류하였을 때 mecA형이 14주, mecR1형 10주, mecA-mecR1형 7주, mecA-mecR1-mecI형이 1주였다. mecA 및 mecA 관련 유전자와 항생제 다제내성과는 상관관계가 없었다. 4) 16주가 mecA와 femA를 모두 보유하였으며, 이들의 plasmid 양상은 I형이 7주, II형이 6주 그리고 III형이 2주였다. Oxacillin 및 vancomycin에 내성을 보이는 II형과 III형에서는 35.5kb의 plasmid DNA를 보유하였다. 5) 장독소 A, B, D, 및 E의 유전자는 51주 전부에서 검출되지 않았으나 장독소 C의 유전자는 64.7%, TSST-1 유전자는 80.4%가 양성이었다. 6) Coagulase 양성이며 mecA 및 femA 유전자를 보유한 MRSA 12균주에 대한 유전자 형별 검사에서 I형에서 X형으로 분류할 수 있었으며, II형 2주, X형 2주가 동일 유전자형이었고, 나머지는 다른 유전자형이었다. 결 론 : 입원 중인 신생아의 비강 내 S. aureus의 상재률이 아주 높았으며, 이들의 항생제 내성률, 특히 vancomycin의 내성률이 높아 이들에 의한 원내 감염에 대한 특별한 관리가 필요하며, 이들의 병원성 인자 및 관련 유전자에 대한 분석은 원내 감염에 대한 적절한 대책과 효과적인 치료에 많은 도움을 줄 수 있으리라 사료된다.

• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.388-398
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• 1998

A bacterium which is resistant to both mercury and cadmium, and also capable of utilizing phenol as a carbon and energy source, was isolated from the Kumho River sediments near Kangchang Bridge, Taegu, Korea. The isolate was labeled Pseudomonas sp. KM10 and characterized. The bacteria grew in 4 mM $CdCl_2$and in $70{\mu}M$ $HgCl_2$. The bacteria efficiently removed over 90% of 1 g/l phenol within 30 h. In the presence of 1.250 g/l phenol, the growth of the microorganism was slightly retarded and the microorganism could not tolerate 1.5 g/l phenol. Curing of plasmid from the bacteria was carried out to generate a plasmidless strain. Subsequent experiments localized the genes for phenol degradation in plasmid and the genes for mercury resistance and cadmium resistance on the chromosome. Dot hybridization and Southern hybridization under low stringent conditions were performed to identify the DNA homology. These results showed significant homologies between the some sequence of the chromosome of Pseudomonas sp. KM10 and merR of Shigella flexneri R 100, and between the some sequence of the chromosome of Pseudomonas sp. KM10 and cadA of Staphylococcus aureus pI258. The mechanism of cadmium resistance was efflux, similar to that of S. aureus pI258 cadA, and the mechanism of mercury resistance was volatilization, similar to that of S. flexneri R100 mer.

• Journal of Microbiology and Biotechnology
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• 제32권12호
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• pp.1537-1546
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• 2022

Staphylococcus aureus is a cause of high mortality in humans and therefore it is necessary to prevent its transmission and reduce infections. Our goals in this research were to investigate the frequency of methicillin-resistant S. aureus (MRSA) in Taif, Saudi Arabia, and assess the relationship between the phenotypic antimicrobial sensitivity patterns and the genes responsible for resistance. In addition, we examined the antimicrobial efficiency and application of silver nanoparticles (AgNPs) against MRSA isolates. Seventy-two nasal swabs were taken from patients; MRSA was cultivated on Mannitol Salt Agar supplemented with methicillin, and 16S rRNA sequencing was conducted in addition to morphological and biochemical identification. Specific resistance genes such as ermAC, aacA-aphD, tetKM, vatABC and mecA were PCR-amplified and resistance plasmids were also investigated. The MRSA incidence was ~49 % among the 72 S. aureus isolates and all MRSA strains were resistant to oxacillin, penicillin, and cefoxitin. However, vancomycin, linezolid, teicoplanin, mupirocin, and rifampicin were effective against 100% of MRSA strains. About 61% of MRSA strains exhibited multidrug resistance and were resistant to 3-12 antimicrobial medications (MDR). Methicillin resistance gene mecA was presented in all MDR-MRSA strains. Most MDR-MRSA contained a plasmid of > 10 kb. To overcome bacterial resistance, AgNPs were applied and displayed high antimicrobial activity and synergistic effect with penicillin. Our findings may help establish programs to control bacterial spread in communities as AgNPs appeared to exert a synergistic effect with penicillin to control bacterial resistance.

• 한국미생물·생명공학회지
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• 제46권3호
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• pp.201-209
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• 2018

Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus fermentum and Lactobacillus casei were successfully isolated from indigenous Indonesian fresh milk based on the general morphological and biochemical classification as described in Bergey's manual. Verification was conducted by sequencing of 16S rRNA after selection using the classification method mentioned in the manual. All isolates exhibited antimicrobial activity against Escherichia coli and Staphylococcus aureus in the well diffusion test. The susceptibilities of the isolated S. thermophilus 24/S1 and L. delbrueckii 94/L4 against 22 different antibiotics were determined by the disc diffusion method and variable susceptibility patterns were observed. Both isolates were susceptible to amoxicillin, the most commonly prescribed antibiotic, and resistant to sulfonamide. The presence of a plasmid was not detected after extraction. S. thermophilus 24/S1 and L. delbrueckii 94/L4 starter cultures were prepared for yogurt production after 9.5 h of incubation and the yogurt was evaluated for its flavor and quality by 30 volunteers. A score of $4.93{\pm}0.45$ out of 7 was obtained as compared to the yogurt prepared using commercial starter cultures which yielded a score of $4.76{\pm}0.30$ out of 7.