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Development of strain-specific polymerase chain reaction primers to detect Fusobacterium hwasookii strains

  • Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.46 no.4
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    • pp.155-159
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    • 2021
  • This study aimed to develop strain-specific polymerase chain reaction (PCR) primers to detect Fusobacterium hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1256, F. hwasookii KCOM 1258, and F. hwasookii KCOM 1268 on the basis of nucleotide sequences of a gene specific to each strain. The unique genes for each F. hwasookii strain were determined on the basis of their genome sequences using Roary. The strain-specific PCR primers based on each strain-specific gene were designed using PrimerSelect. The specificity of each PCR primer was determined using the genomic DNA of the 5 F. hwasookii strains and 25 strains of oral bacterial species. The detection limit and sensitivity of each strain-specific PCR primer pair were determined using the genomic DNA of each target strain. The results showed that the strain-specific PCR primers correspond to F. hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1258, F. hwasookii KCOM 1256/F. nucleatum subsp. polymorphum KCOM 1260, or F. hwasookii KCOM 1268/Fusobacterium sp. oral taxon 203 were developed. The detection limits of these strain-specific PCR primers ranged from 0.2 to 2 ng of genomic DNA for each target strain. The results suggest that these strain-specific PCR primers are valuable in quality control for detecting specific F. hwasookii strains.

Chromosomal studies on the varieties and Formae specials of Fusarium oxysporum.(I) (Fusarium oxysporum의 변종 및 품종의 염색체에 관한 연구( I ))

  • Min, Byung-Re
    • The Korean Journal of Mycology
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    • v.16 no.3
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    • pp.157-161
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    • 1988
  • The vegetative nuclear divisions in hyphae and chromosome numbers were studied with the aid of Giemsa-HCl techniques from 10 strains of Fusarium oxysporum. The entire nuclear division process occurred within an intact nuclear envelope like other fungus. The results confirmed that 2 strains(F. oxysporum S Hongchun D2, F. oxysporum S Jinyang 4) were n=4; 3 strains(F. oxysporum f. sp. lini KFCC 32585, F. oxysporum f. sp. melongenae KFCC 34743 and F. oxysporum f. sp. raphani) n=5; 2 strains(F. oxysporum f. sp. vasinfectum, and F. oxysporum f. sp. mori KFCC 34742) n=6; 3 strains(F. oxysporum f. sp. cucumerium, F. oxysporum f. sp.niveum, and F. oxysporum f. sp. pisi) n=7.

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미생물을 이용한 원유 및 원유제품의 분해 특성

  • O, Gyeong-Taek;Park, Gwi-Hwan;Lee, Jeong-Il;Lee, Jung-Gi;Kim, Seong-Jun;Motoki, Kubo;Jeong, Seon-Yong
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.435-438
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    • 2000
  • Crude oil-degrading microorganisms, Acinetobacter sp. A132, Pseudomonas aeruginosa F722, and Acinetobacter calcoaceticus OM1 were isolated from soil and sea. The optimal temperature of strain A132 and strain F722 on growth isolated from soil was $35^{\circ}C$ both, and also their growth were optimized at pH 8 and 9, respectively. The growth of the strains, A132 and F722, showed that crude oil of 2% (w/v) in culture broth in which crude oil was used as carbon and energy sources appeared to be an optimum. Optimal culture conditions of strain OM1 were different from those of the soil microorganisms except for temperature. The growth of strain OM1 was optimized at pH 7 and crude oil of 3.0% (w/v). The degradability to crude oil by strain A132 showed maximum $5.49g/\;l\;{\cdot}\;day$ under the conditions of $25^{\circ}C$, NaCl of 1.0% (w/v), and crude oil of 2.0% (w/v). The highest degradability of strain F722 to crude oil was $1.19g/\;l\;{\cdot}\;day$ under the culture conditions at $35^{\circ}C$, NaCl 1.0% (w/v), and crude oil of 2.0% (w/v). The degradation characteristics of kerosene $(nC_9-nC_{20})$ and diesel $(nC_9-nC_{28})$ by strain OM1, and F722 were analyzed by gas chromatography. Strain OM1 degraded more than 95% of kerosene and 75% of diesel for 7 days cultivation. Strain F722 showed degradation of more than 80% to kerosene in 10 days.

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Construction of Yeast Strain Suitable for Bioethanol Production by Using Fusion Method (융합법을 이용한 바이오에탄올 생산에 적합한 효모균주의 구축)

  • Kim, Yeon-Hee
    • Journal of Life Science
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    • v.29 no.3
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    • pp.376-381
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    • 2019
  • To construct useful yeast strain for bioethanol production, we improved yeast harboring various phenotypes by using yeast protoplast fusion method. In this study, S. cerevisiae BYK-F11 strain which have ethanol tolerance, thermotolerance and ${\beta}-glucanase$ activity and P. $stipitis{\Delta}ura$ strain which has xylose metabolism pathway were fused by genome shuffling. P. $stipitis{\Delta}ura$ strain was constructed for protoplast fusion by URA3 gene disruption, resulting in uracil auxotroph. By protoplast fusion, several fused cells were selected and BYKPS-F8 strain (fused cell) showing both karyotypes from two parent strains (S. cerevisiae BYK-F11 and P. $stipitis{\Delta}ura$ strain) among 22 fused cells was finally selected. Sequentially, various phenotypes such as ${\beta}-glucanase$ activity, xylose utility, ethanol tolerance, thermotolerance and ethanol productivity were analyzed. The BYKPS-F8 strain obtained ${\beta}-glucanase$ activity from BYK-F11 strain and 1.2 fold increased xylose utility from P. $stipitis{\Delta}ura$ strain. Also, the BYKPS-F8 strain showed thermotolerance at $40^{\circ}C$ and increased ethanol tolerance in medium containing 8% ethanol. In this fused cell, 7.5 g/l ethanol from 20 g/l xylose was produced and the multiple phenotypes were stably remained during long term cultivation (260 hr). It was proved that novel biological system (yeast strains) is easily and efficiently bred by protoplast fusion among yeasts having different genus.

A Brucella Omp16 Conditional Deletion Strain Is Attenuated in BALB/c Mice

  • Zhi, Feijie;Fang, Jiaoyang;Zheng, Weifang;Li, Junmei;Zhang, Guangdong;Zhou, Dong;Jin, Yaping;Wang, Aihua
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.6-14
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    • 2022
  • Brucella spp. are facultative intracellular pathogens that invade, survive and proliferate in numerous phagocytic and non-phagocytic cell types, thereby leading to human and animal brucellosis. Outer membrane proteins (Omps) are major immunogenic and protective antigens that are implicated in Brucella virulence. A strain deleted of the omp16 gene has not been obtained which suggests that the Omp16 protein is vital for Brucella survival. Nevertheless, we previously constructed an omp16 conditional deletion strain of Brucella, ∆Omp16. Here, the virulence and immune response elicted by this strain were assessed in a mouse model of infection. Splenomegaly was significantly reduced at two weeks post-infection in ∆Omp16-infected mice compared to infection with the parental strain. The bacterial load in the spleen also was significantly decreased at this post-infection time point in ∆Omp16-infected mice. Histopathological changes in the spleen were observed via hematoxylin-eosin staining and microscopic examination which showed that infection with the ∆Omp16 strain alleviated spleen histopathological alterations compared to mice infected with the parental strain. Moreover, the levels of humoral and cellular immunity were similar in both ∆Omp16-infected mice and parental strain-infected mice. The results overall show that the virulence of ∆Omp16 is attenuated markedly, but that the immune responses mediated by the deletion and parental strains in mice are indistinguishable. The data provide important insights that illuminate the pathogenic strategies adopted by Brucella.

Biodegradation of Crude Oil and Petroleum Products by Crude Oil-degrading Microorganism (미생물을 이용한 원유 및 원유제품의 분해 특성)

  • 정선용;오경택;박귀환;이정일;이중기
    • KSBB Journal
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    • v.17 no.3
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    • pp.247-254
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    • 2002
  • Two kinds of crude oil-degrading microorganisms from soil and one kind from sea were isolated and named strain Al32, strain F722 and strain OM1, respectively. These microorganism were identified Acinetobacter sp., Pseudomonas aeruginosa and Acinetobacter calcoaceticus, respectively. The optimum cultivation temperature of Acinetobacter sp. A132 and P. aeruginosa F722 was $35^{\circ}C$ and optimum growth pH was 8 and 9, respectively. The growth was the highest at 2.0% (w/v) substrate concentration when crude oil was only carbon source. The growth of A. calcoaceticus OM1 isolated from sea was the highest at 3.0% (w/v) of crude oil. In inspection of crude oil degradability, strain Al32 showed 5.49 g/L.day with Eleuthera (OMAN), 2.0% (w/v). P. aeruginosa F722 showed 1.19 g/L g/L.day with L-Zakum (AFRICA). In case of kerosene $nC_9\simnC_{20}$ and diesel $nC_9\simnC_{28}$, A. calcoaceticus OM1 was degraded 95% and 75%, respectively, for 7 days culture, and P. aeruginosa F722 was 80% after 10 days.

Genome-based identification of strain KCOM 1265 isolated from subgingival plaque at the species level

  • Park, Soon-Nang;Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.45 no.2
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    • pp.70-75
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    • 2020
  • The aim of this study was to identify strain KCOM 1265 isolated from subgingival plaque at the species level by comparing 16S ribosomal RNA gene (16S rDNA) and genome sequences. The whole genome of strain KCOM 1265 was extracted using the phenol-chloroform extraction method. 16S rDNA was amplified using polymerase chain reaction and sequenced using the dideoxy chain termination method. Pairwise genome comparison was performed using average nucleotide identity (ANI) and genome-to-genome distance (GGD) analyses. The data showed that the percent similarity of 16S rDNA sequence of strain KCOM 1265 was 99.6% as compared with those of Fusobacterium polymorphum ATCC 10953T and Fusobacterium hwasookii KCOM 1249T. The ANI values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 95.8% and 93.0%, respectively. The GGD values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 63.9% and 49.6%, respectively. These results indicate that strain KCOM 1265 belongs to F. polymorphum.

Surface Strain Effects on the Adsorption of Au Adatoms on MgO(001) Surfaces with Surface O Vacancies

  • Jeon, Junjin;Park, Jinwoo;Yu, Byung Deok
    • Journal of the Korean Physical Society
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    • v.73 no.9
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    • pp.1324-1328
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    • 2018
  • By employing ab-initio total-energy and electronic-structure calculations based on the density-functional theory, we studied the effects of surface strain ${\varepsilon}_s$ on the adsorption properties of a Au adatom on defective MgO(001) surfaces with surface oxygen vacancies ($F_s$ centers). The formation energy of the $F_s$ center on MgO(001) varied very slightly in the region of ${\varepsilon}_s$ from -6% to -4% and monotonically decreased with the increase in ${\varepsilon}_s$, from -4% to +6%. As ${\varepsilon}_s$ increased, the adsorption energy ($E^{Fs}_{ads}$) of Au on the $F_s$ center of strained MgO(001) monotonically decreased and, in particular, showed a much larger decrease in $E^{Fs}_{ads}$ for a tensile surface strain of ${\varepsilon}_s$ > +4%. The surface strain dependence on the physical properties, such as the charge states, the spatial charge rearrangement, for Au on the $F_s$ center of strained MgO(001) surfaces was also analyzed. These results provide important physical information on the effects of surface strain on the adsorption of Au on MgO(001) surfaces with $F_s$ centers.

Flavobacterium jocheonensis sp. nov., Isolated from Marine Green Alga Ulva pertusa

  • Choi, Ha Ri;Park, So Hyun;Heo, Moon Soo
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1266-1272
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    • 2019
  • A bacterial strain, labeled $UR11^T$, was isolated from green alga Ulva pertusa collected from Jeju Island, Korea. $UR11^T$ was identified as a gram-negative, rod-shaped, motile by gliding and aerobic bacterial strain with yellow colonies on R2A plates. The strain $UR11^T$ grew over at a temperature range of $10^{\circ}C$ to $30^{\circ}C$ (optimally at $25^{\circ}C$), a pH range of 6.0-11 (optimally at pH 7.0) and a Nacl range of 0.5-5% Nacl (w/v). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain $UR11^T$ was a member of the genus Flavobacterium. Strain $UR11^T$ shared close similarity with F. jejuensis $EC11^T$ (98.0%) F. jumunjinense $HME7102^T$ (96.1%), F. haoranii $LQY-7^T$ (95.3%), F. dongtanense $LW30^T$ (95.1%), and F. ahnfeltiae 10Alg $130^T$(94.9%). The major fatty acids (>5%) were $iso-C_{15:0}$ (33.9%), $iso-C_{15:1}$ G (12.4%), $iso-C_{17:0}$ 3-OH (9.0%), $isoC_{16:0}$ (7.0%) and $iso-C_{15:0}$ 3-OH (6.3%). The major polar lipids were phosphatidylethanolamine, seven unknown aminolipids, two unknown aminopolarlipids and two unknown lipids. DNA-DNA hybridization value was 58% at strain $UR11^T$ with F. jejuensis $EC11^T$. Based on phenotypic, chemotaxonomic and phylogenetic evidence, strain $UR11^T$ represents a novel species of the genus Flavobacterium, for which the name Flavobacterium jocheonensis sp. nov. is proposed. The type strain is Flavobacterium jocheonensis is $UR11^T$ (=KCTC $52377^T$ =JCM $31512^T$).

Numerical Prediction of NOx in the Nonpremixed Hydrogen-Air Flame using the Quasi-Laminar Reaction Modelling (준충류 근사를 이용한 수소-공기 비예혼합화염의 질소산화물 생성예측)

  • Kim, Seong-Lyong;Jeung, In-Seuck;Yoon, Young-Bin
    • Journal of the Korean Society of Combustion
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    • v.4 no.1
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    • pp.131-139
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    • 1999
  • A Numerical Analysis of NOx production in Hydrogen-Air flame is performed using the quasi-laminar reaction modelling. As results, in low global strain rate region, $U_F/D_F\;{\leq}\;50,000$, the quasi-laminar reaction modelling reproduces the experimentally observed EINOx half power scaling that the ratio of EINOx and flame residence time, $L_f^3(D_F^2U_F)$, is proportional to the square root of global strain rate. Thus, it suggests that turbulence-chemistry interaction has a minor impact on the trend of NOx production in low global strain rate region. However, the quasi-laminar reaction modelling predicts the higher temperature and NOx than experimentally observed. This overprediction may be due to the lack of radiation and quasi-laminar reaction modelling.

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