• Title/Summary/Keyword: isolation techniques

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Artificial induction and isolation of cadmium-tolerant soil bacteria

  • Lee, Sangman
    • Journal of Applied Biological Chemistry
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    • v.63 no.2
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    • pp.125-129
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    • 2020
  • Environmental pollution caused by various heavy metals is a serious global problem. To solve this problem, microbial bioremediation of contaminated metals has developed rapidly as an effective strategy when physical and chemical techniques are not suitable. In this study, cadmium (Cd)-tolerant soil bacteria were isolated via artificial induction in laboratory conditions instead of screening bacteria naturally adapted to metal-contaminated soils. Wild-type (WT) bacteria grown in uncontaminated soils were artificially and sequentially adapted to gradually increasing Cd concentrations of up to 15 mM. The resultant cells, named Soil-CdR15, survived at a Cd concentration of 10 mM, whereas WT cells failed to survive with 4 mM Cd on solid media for 2 d. In liquid media containing Cd, the SoilCdR15 cells grew with 15 mM Cd for 7 d, whereas the WT cells could not grow with 5 mM Cd. Both Soil-CdR15 and WT cells removed approximately 35% of Cd at the same capacity from liquid media containing either 0.5 or 1.0 mM Cd over 2 d. In addition to Cd, the Soil-CdR15 cells showed increased resistance to nickel, zinc, and arsenic compared to WT cells. The Soil-CdR cells were identified as Burkholderia sp. by partial sequencing of 16S rRNA. The data presented in this study demonstrate that isolation of heavy metal-tolerant microorganisms via artificial induction in laboratory conditions is possible and may be useful for the application of the microorganisms for the bioremediation of heavy metals.

Isolation and Identification of Yeast Strain from Fermented Tea (발효차로부터 효모의 분리 및 동정)

  • Kang, Ok-Ju
    • Korean journal of food and cookery science
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    • v.24 no.1
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    • pp.11-15
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    • 2008
  • In searching for yeast to be utilized as biocontrol agents, a single yeast strain was isolated from Camellia sinensis based on its morphological, cultural, physiological, and biochemical properties, as well as by molecular techniques. This single strain was pink to red in color and designated as strain JY-1. The effects of temperature, pH, NaCl concentration, and ethanol concentration on the growth of the JY-l strain were examined for the JY-1. Growth occurred at temperatures ranging from 20 to $35^{\circ}C$, and between pH 3.0 and 12.0, with optimal growth at $25-30^{\circ}C$ and pH 5.0. The yeast also grew in the presence of 0-2% (w/v) NaCl and 0-4% (v/v) EtOH. The isolate was further classified based on biochemical characteristics using the VITEK system. The biochemical data obtained using this system were similar to those of Rhodotorula glutinis/Rhodotorula mucilaginosa (exhibiting a 93% matching level). Molecular phylogenetic analysis based on l8S rDNA sequences indicated that the yeast represented a basidiomycetous species, and its highest degree of sequence similarity was with Rhodosporidium azoricum, strain JCM11251 (99%).

Identification of a High-yield Technique for Isolating Endometrial Epithelial Cells from the Mouse Uterus : A Comparison of Mechanical and Sedimentation-adherence Methods

  • Sohn, Jie Ohn;Jo, Yoon Mi;Park, Hye Jin;Ahn, Ji Yeon;Song, Hyun Jin;Lim, Jeong Mook;Lee, Seung Tae
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.73-80
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    • 2016
  • An in vitro assay following culture of endometrial epithelial cells is essential for understanding epithelial cell function in reproduction. Several diverse techniques have been developed for isolating endometrial epithelial cells, although an optimal technique has not been identified. In this study, we describe a sedimentation-adherence (S-A) isolation technique with a high-yield cell-separating ability to isolate endometrial epithelial cells from 8-week-old female C57BL/6 mice. We analyzed total cell number, viability, morphology, and expression of cytokeratin 18 as an endometrial epithelial cell-specific marker in cells isolated using a mechanical method compared to the S-A technique. There were no significant differences in the total number, viability, or morphology of the putative endometrial epithelial cells with either method. In contrast, significantly more endometrial epithelial cells harvested using the S-A method were positively stained for cytokeratin 18 than those isolated using the mechanical method. These results confirm that the S-A method is more efficient for retrieving endometrial epithelial cells than a mechanical method.

Application of Molecular Methods for the Identification of Acetic Acid Bacteria Isolated from Blueberries and Citrus Fruits

  • Gerard, Liliana Mabel;Davies, Cristina Veronica;Solda, Carina Alejandra;Corrado, Maria Belen;Fernandez, Maria Veronica
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.193-204
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    • 2020
  • Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.

A Study on Real Time Fault Diagnosis and Health Estimation of Turbojet Engine through Gas Path Analysis (가스경로해석을 통한 터보제트엔진의 실시간 고장 진단 및 건전성 추정에 관한 연구)

  • Han, Dong-Ju
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.49 no.4
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    • pp.311-320
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    • 2021
  • A study is performed for the real time fault diagnosis during operation and health estimation relating to performance deterioration in a turbojet engine used for an unmanned air vehicle. For this study the real time dynamic model is derived from the transient thermodynamic gas path analysis. For real fault conditions which are manipulated for the simulation, the detection techniques are applied such as Kalman filter and probabilistic decision-making approach based on statistical hypothesis test. Thereby the effectiveness is verified by showing good fault detection and isolation performances. For the health estimation with measurement parameters, it shows using an assumed performance degradation that the method by adaptive Kalman filter is feasible in practice for a condition based diagnosis and maintenance.

Development of Cell Lines for Application of Recombinant DNA Techniques in Crops (작물의 유전자 재조합을 위한 세포주의 개발 연구)

  • Chae, Young-Am;Choi, Kyu-Whan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.2
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    • pp.195-200
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    • 1985
  • This experiment was carried out to know the processes of protoplast isolation, culture and plant regeneration in aims of introducing foreign genes into plant cells through plant gene vector, and cellular selection for plant improvement. The main results indicated that 2% cellulase plus 0.5% macerozyme is proper for isolation of protoplasts from leaf mesophyll cells of N. plumbaginifolia, plating efficiency was higher in 1.4-2.0 x 10$^4$ cells/ml, complete cell wall was regenerated after 2 days culture, cell division and cell mass were observed after 4 days and 2 weeks, respectively, colony was developed after 3 weeks culture, addition of 1-2mg/l BA promoted shoot differentiation while root differentiation did not required hormone and seeds were harvested from more than 100 cell lines for further investigation and study.

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Isolation and Chemical Structure Identification of Allelopathic Substances from the Ginkgo(Ginkgo biloba L.) Leaf Waste Produced by Phamaceutical Process (제약폐기 은행잎중에 함유된 식물생육억제물질 분리동정)

  • Seong, Ki-Seog;Kim, Bok-Jin;Kwon, Oh-Kyung;Choi, Du-Hoi
    • Korean Journal of Soil Science and Fertilizer
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    • v.30 no.4
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    • pp.377-383
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    • 1997
  • Studies were conducted on the nature of allelopathic effect of the substances in the waste of ginkgo leaves from pharmaceutic factory. In the first step, to find out whether there was any allelophatic effect, young seedlings of radish and rice were grown in the water (crude) extract of ginkgo leaf waste and in different liquid/lquid partitioned fractions of EtOAc at pH 9, EtOAc at pH 3, and BuOH. As second step, attempts were made to isolate and identify the allelophatic substance in different liquid/liquid partitioned fractions using GC/MS and NMR techniques. The water (crude) extract of ginkgo leaf waste retarded the growth of radish seedlings under 10% concentration. In case of rice seedlings, the water extract of ginkgo leaf extract showed adverse effect on the growth when combined with $3.3{\times}10^{-6}M$ gibberellin A3. All of the liquid/liquid fractions of crude extract showed strong retardation of seedling growth of radish and rice at the concentration of 1%. Allelophatic substance was isolated from the crude extract using liquid/liquid partition, column chromatography and HPLC techniques. The analytical results of isolated componet using GC/MS and NMR proved that the allelophatic substance in the ginkgo leaf wastes is catechol; one of phenol compounds. Based on the experiences current study, a practical method for the testing of allelophatic effect of crude extract of some materials was proposed. In this method, rice seeds were allowed to sprout until the length of coleoptile to reach 0.5 mm. Such seedlings were submerged in the solution containing supposedly allelophatic substance and the length of shoot and root was measured 3 days after treatment.

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The Effect of Papaverine on the Calcium-dependent $K^+$ Current in Rat Basilar Smooth Muscle Cells

  • Bai, Guang-Yi;Cho, Jae-Woo;Han, Dong-Han;Yang, Tae-Ki;Gwak, Yong-Geun;Kim, Chul-Jin
    • Journal of Korean Neurosurgical Society
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    • v.38 no.5
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    • pp.375-379
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    • 2005
  • Objective : Papaverine has been used in treating vasospasm following subarachnoid hemorrhage[SAH]. However, its action mechanism for cerebral vascular relaxation is not clear. Potassium channels are closely related to the contraction and relaxation of cerebral smooth muscle. Therefore, to identify the role of potassium and calcium channels in papaverine-induced vascular relaxation, we examine the effect of papaverine on potassium channels in freshly isolated smooth muscle cells from rat basilar artery. Methods : The isolation of rat basilar smooth muscle cells was performed by special techniques. The whole cell currents were recorded by whole cell patch clamp technique in freshly isolated smooth muscle cells from rat basilar artery. Papaverine was added to the bath solution. Results : Papaverine of $100{\mu}M$ into bath solution increased the amplitude of the outward $K^+$ current which was completely blocked by BKCa[large conductance calcium dependent potassium channels]blocker, IBX[iberiotoxin], and calcium chealator, BAPTA[l,2-bis[o-aminophenoxy]ethane-N,N,N',N'-tetraacetic acid], in whole cell mode. Conclusion : These results strongly suggest that potassium channels may play roles in papaverine-induced vascular relaxation in rat basilar artery.

Two-Failure Gps Raim by Parity Space Approach (패러티 공간을 이용한 2개 GPS 파라미터 고장진단)

  • Yoo, Chang-Sun;Ahn, Iee-Ki;Lee,Sang-Jeong
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.31 no.6
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    • pp.52-60
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    • 2003
  • In aviation navigation using GPS, requirements on availability and integrity must be absolutely satisfied. Current study on accomplishing this integrity includes RAIM(Receiver Autonomous Integrity Monitoring), monitoring integrity internaIly in GPS receiver itself. Parity space technique as one of RAIM techniques has shown the advantages in fault detection and isolation due to each use of its magnitude and direction under the assumption of one fault. ln case of multiple fault, as biases in errors interact decreasing the effect of multiple fault in parity space, the exact fault detection and identification(FDI) may be difficult to be conducted. This paper focuses on FDI study on two faults and explains why parity space techniques applied on single fault is not adequate to the application of multiple fault case and shows that extended parity space technique may improve the performance of RAIM on two faults.

Redundancy Management Design for Triplex Flight Control System (3중 비행제어시스템의 다중화 기법 설계)

  • Park, Sung-Han;Kim, Jae-Yong;Cho, In-Je;Hwang, Byung-Moon
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.38 no.2
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    • pp.169-179
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    • 2010
  • Satisfying the same probability of loss of control and essentially two fail operative performance with a triplex computer architecture requires a lot of modification of the conventional redundancy management design techniques, previously employed in quadruplex digital flight control computer. T-50 FCS for triplex redundancy management design applied an advanced digital flight control architecture with an I/O controller which is functionally independent of the digital computer to achieve the same reliability and special failure analysis and isolation schemes for fail operational goals with a triplex configuration. The analysis results indicated that the triplex flight control system is to satisfy the safety requirement utilizing the advanced flight control techniques and the system performance of the implemented flight control system was verified by failure mode effect test.