• Title/Summary/Keyword: strain SK1

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

  • Kang, Hyun
    • Biomedical Science Letters
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    • v.1 no.1
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    • pp.55-72
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    • 1995
  • Forty gentamicin-resistant isolates of Enterococcus faecalis were selected from various clinical materials, determined their antimicrobial susceptibility, and studied there R-plasmid characteristics and polypeptide patterns. All of the isolates were susceptible to vancomycin. The MICs($\mu$/ml) of antimicrobial agents to the isolates were as follows; the MIC of gentamicin was 128 and $\geq$2040, ampicillin 1 and 1, chlorarmphenicol 2 and 8, erythromycin 32 and 256, and vancomycin 1 and 2. E. faecalis HL-1 strain had 8 plasmid DNA elements, HL-2 and HL-3 strains had 6, HL-4 had 7, HL-5 had 4, and HL-6 had 5. The 51.7 Kb of gentamicin resistance plasmid DNA was conjugally transferred from two strains of E. faecalis HL-1 and HL-6 to S. aureus SK 982. The plasmid transfer frequency between S. aureus SK 982 and E. faecalis HL-1 or E. faecalis HL-6 was 6.3$\times10^{-4} and 3.7$\times10^{-5}$, respectively. Plasmid curing ratio after the treatment of ethidium bromide(10$\mu$/ml) to E. faecalis tarnsconjugants R-1 and R-6 were about 51% and 67%, respectively. The tetracycline gene was located in 2.15 Kb plasmid of E. faecalis HL-1, but it was not found in the E. faecalis HL-6 by Southern blot analyses. The antigenic components of E. faecalis HL-1, HL-6, R-1 and R-6 strains were analyzed by SDS-PAGE and immunoblotting. The E. faecalis strains had 7 to 16 polypeptide bands, however their major proteins were 97.8 and 26.8 Kd. At the Immunoblotting, 97.8, 95.8, 74.8, 63.5, 33.7 and 26.8 Kd polypeptides of the strains showed major antigenic activities with patient's sera infected intra-abdominally with an E. faecalis strain.

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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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    • v.16 no.2
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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.

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

  • Choi Jeong-Wha;Kim Myung-Ju;Lee Joo-Young
    • Journal of the Korean Society of Clothing and Textiles
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    • v.29 no.8 s.145
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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 ($616cm^2$).

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

  • 길영식;신희정;김영창
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.46-51
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    • 2000
  • Salmonella typhi is one of important causes of human enteric infections. S. typhi KNIH100 was isolated from a patient of typhoid fever in Korea. We cloned a 5.0 kb SalⅠ fragment containing the aroA gene encoding a 5-enolpyruvylshikimate-3-phosphate synthetase from chromosomal DNA of this strain. This recombinant plasmid was named pSAL80. E. coli CGSC2829, an aroA- mutant, was not grown on the M9 minimal medium but E. coli CGSC2829 (pSAL80) was grown on the M9 minimal medium. The aroA gene was composed of 1,284 base pairs with ATG initiation codon and TAA termination codon. Sequence comparison of the aroA gene exhibited 99%, 98%, and 77% identity with those of S. typhi Ty2, S. typhimurium, and E. coli respectively. As in the cases of Shigella sonnei and E. coli, the serC and aroA genes lie in a single operonic structure.

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

  • 최정화;이주영
    • Journal of the Korean Society of Clothing and Textiles
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    • v.26 no.11
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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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    • v.37 no.3
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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 $60^{\circ}C$.

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

  • Choi, Jeong-Wha;Park, Joon-Hee
    • Journal of the Korean Society of Clothing and Textiles
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    • v.31 no.6 s.165
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    • pp.955-965
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    • 2007
  • Safety helmets are used widely in various industries by workers since they are legally required to wear them. However, thermal discomfort is one of the major complaints from helmet users. To relieve this problem, frozen gelled packs can be considered for use inside the helmets. In this paper, tests were performed on humans to evaluate the physiological strains of wearing safety helmets and to investigate the effects of using frozen gelled packs inside the helmets. Experiments were conducted in a climatic chamber of WBGT $33{\pm}1^{\circ}C$ under four differed experimental conditions: 1) not wearing a safety helmet(NH); 2) wearing a safety helmet with frozen gelled pack A(HA); 3) wearing a safety helmet with frozen gelled pack B(HB); and 4) wearing only a safety helmet(OH). The results were as follows. First, when comparing NH with OH, physiological responses such as $\bar{T}_{sk},\;T_r$, HR and sweat rate were significantly higher in OH and subjective sensations were reported as less hot and more comfortable than NH(p<.05). Second, in regard to the frozen gelled packs inserted inside the safety helmets, some physiological responses in HA were different from those in HB, according to the two different types of packs. HA was hotter, more uncomfortable and less exhausted than HB. However, result from both HA and HB were lower than those from OH in terms of temperature and humidity inside safety helmet, sweat rate, $T_r$ increase, heat storage(p<.05). When wearing safety helmets with frozen gelled packs, it was shown that heat strain can be alleviated. These results are expected to help millions of workers who complain that wearing safety helmets is uncomfortable and messy.