• 제목/요약/키워드: strain SK1

검색결과 57건 처리시간 0.026초

Gentamicin 고도내성 Enterococcus faecalis균주의 항균제감수성, R-플라스미드 및 항원의 특성연구 (Studies on Antimicrobial Susceptibility and Characteristics of R-plasmids and Antigens of High-level Gentamicin Resistant Enterococcus faecalis)

  • 강현
    • 대한의생명과학회지
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    • 제1권1호
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    • pp.55-72
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    • 1995
  • 임상 검체에서 분리 동정한 E.faecalis 균주에 대하여 gentamicin및 타 항균제에 대한 감수성을 조사했고, R-플라스미드 DNA패턴 및 제한효소에 의한 DNA단편분석,filter-mating에 의한 gentamicin 플라스미드 DNA 전이, R-플라스미드 DNA 제거, tetracycline 내성 유전자확인, 항원분석을 실시하였다. 분리된 40주 E. faecalis 모두가 gentamicin에는 내성이었고, 25주는 요 검체에서 분리되었으며, 이 40균주들 중 95%가 입원환자에서 분리되어 중요한 원내감염균임을 알 수 있었다. Gentamicin 고도내성 E. faecalis는 24주(60%)이었고, 최소억제농도의 범위는 ampicillin이 1~64$\mu$g/ml, Chroramphenicol 이 8~128$\mu$g/ml 과 erythromycin 128 $\mu$g/ml, vancomycinol 1~2 $\mu$g이었다. 분리한 Gentamicin 고도내성 E. faecalis HL-1은 4가지 항균제에 모두 내성이었고, 7종류의 플라스미드가 있었다. HL-2와 HL-3은 모두 6종류, HL-4는 7종류, HL-5는 4종류 그리고 HL-6은 5종류의 플라스미드가 있었다. E. faecalis HL-1 과 HL-6의 내성전달 빈도는 6.3$\times10^{-4}$ 과 3.7$\times10^{-5}$이 었으며, E faecalis HL-1의 내성전달 빈도가 높았다 접합에 의해 E.faecalir HL-1과 HL-6의 플라스미드 중 51.7 Kb크기의 전이된 DNA가 확인되어 gentamicin 내성이 플라스미드 전이에 의한 것임을 알 수 있었다. Tetracycline 유전자는 E.faecalis transconjugants R-1의 2.15 Kb 플라스미드에 있었다. 공여균주 및 transconjugants균주의 항원성을 Immunoblotting으로 분석한 결과 E. faecalis HL-1과 E. faecalis transconjugants R-1균주는 97.8, 46.8 Kd의 분자량을 갖는 단백질과 공통적으로 반응하였고, E. faecalis HL-6와 E. faecalis transconjugants R-6균주는 46.8 Kd의 분자량을 갖는 단백질과 공통적으로 반응하였다. 그 반응의 강도는 85.8, 97.8, 46.8, 33.7, 63.5 와 74.8 Kd 순이었다. 공여균주 E. faecalis HL-6에서는 반응하지 않았던 97.8, 95.8, 74.8와 63.5 Kd의 단백질이 E. faecalis transconjugants R-6균 주에서 반응하였다. 이들 종 특이성 단백질을 Invitro에서는 배양되지 않는 E. faecalis에 대한 혈청학적 진단의 항원으로 이용한다면 유용할 것이다.

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Mutation Analysis of Synthetic DNA Barcodes in a Fission Yeast Gene Deletion Library by Sanger Sequencing

  • Lee, Minho;Choi, Shin-Jung;Han, Sangjo;Nam, Miyoung;Kim, Dongsup;Kim, Dong-Uk;Hoe, Kwang-Lae
    • Genomics & Informatics
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    • 제16권2호
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    • pp.22-29
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    • 2018
  • Incorporation of unique barcodes into fission yeast gene deletion collections has enabled the identification of gene functions by growth fitness analysis. For fine tuning, it is important to examine barcode sequences, because mutations arise during strain construction. Out of 8,708 barcodes (4,354 strains) covering 88.5% of all 4,919 open reading frames, 7,734 barcodes (88.8%) were validated as high-fidelity to be inserted at the correct positions by Sanger sequencing. Sequence examination of the 7,734 high-fidelity barcodes revealed that 1,039 barcodes (13.4%) deviated from the original design. In total, 1,284 mutations (mutation rate of 16.6%) exist within the 1,039 mutated barcodes, which is comparable to budding yeast (18%). When the type of mutation was considered, substitutions accounted for 845 mutations (10.9%), deletions accounted for 319 mutations (4.1%), and insertions accounted for 121 mutations (1.6%). Peculiarly, the frequency of substitutions (67.6%) was unexpectedly higher than in budding yeast (~28%) and well above the predicted error of Sanger sequencing (~2%), which might have arisen during the solid-phase oligonucleotide synthesis and PCR amplification of the barcodes during strain construction. When the mutation rate was analyzed by position within 20-mer barcodes using the 1,284 mutations from the 7,734 sequenced barcodes, there was no significant difference between up-tags and down-tags at a given position. The mutation frequency at a given position was similar at most positions, ranging from 0.4% (32/7,734) to 1.1% (82/7,734), except at position 1, which was highest (3.1%), as in budding yeast. Together, well-defined barcode sequences, combined with the next-generation sequencing platform, promise to make the fission yeast gene deletion library a powerful tool for understanding gene function.

여름철 농민의 서열 부담 경감을 위한 냉각조끼의 성능 평가 (Efficacy of Cooling Vests for Alleviating Heat Strain of Farm Workers in Summer)

  • 최정화;김명주;이주영
    • 한국의류학회지
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    • 제29권8호
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    • pp.1176-1187
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    • 2005
  • The purpose of this study was to evaluate the efficiency of cooling vests developed for farm workers harvesting red pepper in summer. The study was performed using the following two steps: 1) Climatic chamber test, 2) Field test. For the chamber test, a work environment was simulated as $33^{\circ}C$ and $65\%$RH, and the thermo-physiological and subjective responses were measured with and without cooling vests. Twelve young males participated as subjects. For the field test, three farmers participated while harvesting red pepper on the form, in summer. The measurements used were same as in the chamber test. Subjects were tested without any cooling vests, as a control. They were tested wearing vests with 2 frozen gel packs (CV2: Cooling area, $308cm^2$), and vests with 4 frozen gel packs (CV4: Sooting area, $616cm^2$). As a result of the chamber test, rectal temperature($T_{re}$) and mean skin temperature( $T_{sk}$) were lower in both CVs than in Control, and this tendency was statistically significant in CV4 (p<.05). Clothing microclimate temperature ($T_{clo}$) and total sweat rate (TSR) were significantly lower when wearing cooling vests (p<.05) Heart rate (HR) was also lower in wearing cooling vests than in Control, and the speed of recovery to the comfort level was faster when the subjects wore cooling vests. In addition, subjects felt 'less hot, less humid, and less uncomfortable' in both CVs than in Control. Field tests showed a similar tendency with the chamber tests. In particular, wearing the cooling vest was effective in restraining the raise of $T_{clo}$ on the back. It can be concluded that the cooling vest was effective in alleviating heat strain and discomfort in both the chamber test and the field test, despite the cooling area of the cooling vest being just $3.4\%$ of the body surface area ($616cm^2$).

Salmonella typhi KNIH100으로부터 aroA 유전자의 클로닝과 염기서열 분석 (Cloning and Nucleotide Sequence Analysis of the aroA Gene from Salmonella typhi KNIH100)

  • 길영식;신희정;김영창
    • 미생물학회지
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    • 제36권1호
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    • pp.46-51
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    • 2000
  • 장티푸스는 Salmonella typhi에 의해 유발되는 장감염성 질환으로 사람과 동물에 공통되는 질병이다. 본 연구에서는 국립보건원과 공동연구를 수행하여 한국형 장티푸스 유발균인 S. typhi KNIH100을 분리하였다. 분리된 S. typhi KNIH100의 염색체 DNA로부터 방향족 아미노산의 생합성에 관여하는 효소인 5-enolpyruvylshikimate-3-phosphate synthetase를 암호화하는 aroA 유전자를 포함하는 약 5.0 kb의 SalI절편을 pBluescriptII SK(+) vector와 aroA 돌연변이주인 E. coli CGSC2829를 이용하여 클로닝하였다. 그리고 이 클론을 pSAL80이라 명명하였다. 클로닝된 재조합 plasmid인 pSAL80에는 ATG 개시코돈과 TGA 종결코돈을 포함하는 1,284 염기로 구성된 aroA 유전자가 위치하고 있었으며, 다른 장내세균과 마찬가지로 serC와 하나의 오페론을 구성하고 있음을 밝혔다. 또한 S. typhi Ty2, S. typhimurium, 그리고 E. coli 등 다른 장내세균의 aroA 유전자와 상동성을 비교하여 본 결과 각각 99%, 95%, 77%의 상동성을 나타내었다.

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농약 방제 작업자의 작업 환경 및 노동 부담 평가 (Evaluation of the thermal environments and the workload of farmers during the spraying pesticide in the rice field)

  • 최정화;이주영
    • 한국의류학회지
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    • 제26권11호
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    • pp.1672-1681
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    • 2002
  • To evaluate the thermal environments and the workload of farmers in the rice field in summer, this study investigated rice farmers' physiological, psychological responses, work postures, work clothes, air temperature and air humidity during the spraying pesticide in the rice field. Five career farmers (3 males, 2 females) volunteered as the subjects. During the spraying pesticide in the rice field, physiological responses were monitored continuously. The results were as follows. l. Farmers wore only raincoats not pesticide-proof clothing. 2. The value of WBGT, rectal temperature($T_{re}$), mean skin temperature(${\={T}}_{sk}$) were $24.9∼28.9^{\circ}C,\;37.8({\pm}0.3)^{\circ}C\;and\;33.6({\pm}0.6)^{\circ}C$, respectively. Clothing microclimate temperature($T_{cl}$) on the chest and back were $32.5({\pm}2.6)^{\circ}C\;and\;33.6({\pm}2.6)^{\circ}C$, respectively(p<0.00l). Humidity inside of the clothing ($H_{cl}$) was over 80%RH and heart rate(HR) was 112(${\pm}27$)bpm. We evaluated that the spraying pesticide was 'heavy work' by the Tre and HR. To four subjective questionnaires, all farmers expressed 'hard, hot, humid and uncomfortable' without individual difference at the end of works. We suggested that 1) the spraying pesticide in the rice field was a heavy work, 2) because the workload of farmers in the raincoat/pesticide-proof clothing can't be evaluated by only WBGT, assessors should measure physiological, psychological responses as well as thermal environments, 3) to alleviate farmers' heat strain, clothing manufacturers must consider not only the improvement of textile materials and clothing weight but also the designing of personal cooling equipment.

Isolation, Characterization, and Molecular Cloning of the cDNA Encoding a Novel Phytase from Aspergillus niger 113 and High Expression in Pichia pastoris

  • Xiong, Ai Sheng;Yao, Quan-Hong;Peng, Ri-He;Li, Xian;Fan, Hui-Qin;Guo, Mei-Jin;Zhang, Si-Liang
    • BMB Reports
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    • 제37권3호
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    • pp.282-291
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    • 2004
  • Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

머리 냉각시의 인체생리반응 - 안전모 착용을 중심으로 - (Physiological Responses of Wearing Safety Helmet with Cooling Pack in Hot Environment)

  • 최정화;박준희
    • 한국의류학회지
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    • 제31권6호
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    • pp.955-965
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    • 2007
  • 안전모는 다양한 산업현장에서 머리 보호를 위해 착용되는 보호구로 법적으로도 착용이 의무화되어있다. 그러나, 안전모 착용에 따른 불쾌감 및 생리적 부담은 작업자들이 착용을 꺼려하는 주된 원인이 된다. 본 연구에서는 이 문제를 해결하기 위한 하나의 방안으로 안전모에 냉매팩을 삽입하는 방법을 제안하였고, 동시에 냉매팩 삽입으로 인한 안전모의 무게 증가가 인체생리반응에 미치는 영향을 규명하고자 하였다. WBGT $33{\pm}1^{\circ}C$의 환경에서 인체착용평가 방법을 이용하여, 4가지 실험조건으로 안전모 착용시의 인체 생리반응을 조사하였고, 그 결과는 다음과 같다. 첫째, 안전모만 착용 시(OH)와 안전모 미착용시(NH)를 비교해 보면, 안전모 착용 시에 평균피부온, 직장온, 심박수, 발한량 등이 통계적으로 유의하게 높았고, 주관적인 감각에서도 더 덥고, 더 습하며, 불쾌하고 주관적으로도 힘들다고 느끼는 것으로 나타났다. 둘째, 냉매팩 삽입시의 경우(HA, HB), 냉매팩의 종류에 따라 여러 인체생리반응에 차이를 보였으나, 안전모만 착용한 경우(OH)와 비교 시 안전모내 기후, 발한량, 직장온 상승도, 열저장(heat storage) 등이 더 낮았다. 한서감과 쾌적감에서는 냉매팩 종류에 따라 다른 경향을 보였다. 본 결과를 통해 안전모 착용시의 생리적, 주관적 부담 정도를 객관적으로 파악할 수 있었으며, 안전모에 냉매팩을 삽입하는 방법을 서열 스트레스를 경감시키는 하나의 방법으로 제시할 수 있었다. 추후 연구에서는 냉매팩의 무게, 피복면적, 성분 등을 고려해야 할 것이며, 본 결과가 서열 환경에서 안전모를 착용하고 작업하는 여러 산업 근로자에게 도움이 되기를 기대한다.