• Title/Summary/Keyword: laboratory analysis

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Proteomic Analysis and Extensive Protein Identification from Dry, Germinating Arabidopsis Seeds and Young Seedlings

  • Fu, Qiang;Wang, Bai-Chen;Jin, Xiang;Li, Hong-Bing;Han, Pei;Wei, Kai-Hua;Zhang, Xue-Min;Zhu, Yu-Xian
    • BMB Reports
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    • v.38 no.6
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    • pp.650-660
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    • 2005
  • Proteins accumulated in dry, stratified Arabidopsis seeds or young seedlings, totaled 1100 to 1300 depending on the time of sampling, were analyzed by using immobilized pH gradient 2-DE gel electrophoresis. The molecular identities of 437 polypeptides, encoded by 355 independent genes, were determined by MALDI-TOF or TOF-TOF mass spectrometry. In the sum, 293 were present at all stages and 95 were accumulated during the time of radicle protrusion while another 18 appeared in later stages. Further analysis showed that 226 of the identified polypeptides could be located in different metabolic pathways. Proteins involved in carbohydrate, energy and amino acid metabolism constituted to about 1/4, and those involved in metabolism of vitamins and cofactors constituted for about 3% of the total signal intensity in gels prepared from 72 h seedlings. Enzymes related to genetic information processing increased very quickly during early imbibition and reached highest level around 30 h of germination.

Expression of $HpaG_{Xooc}$ Protein in Bacillus subtilis and its Biological Functions

  • Wu, Huijun;Wang, Shuai;Qiao, Junqing;Liu, Jun;Zhan, Jiang;Gao, Xuewen
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.194-203
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    • 2009
  • $HpaG_{Xooc}$, from rice pathogenic bacterium Xanthomonas oryzae pv. oryzicola, is a member of the harpin group of proteins, eliciting hypersensitive cell death in non-host plants, inducing disease and insect resistance in plants, and enhancing plant growth. To express and secret the $HpaG_{Xooc}$ protein in Bacillus subtilis, we constructed a recombinant expression vector pM43HF with stronger promoter P43 and signal peptide element nprB. The SDS-PAGE and Western blot analysis demonstrated the expression of the protein $HpaG_{Xooc}$ in B. subtilis. The ELISA analysis determined the optimum condition for $HpaG_{Xooc}$ expression in B. subtilis WBHF. The biological function analysis indicated that the protein $HpaG_{Xooc}$ from B. subtilis WBHF elicits hypersensitive response(HR) and enhances the growth of tobacco. The results of RT-PCR analysis revealed that $HpaG_{Xooc}$ induces expression of the pathogenesis-related genes PR-1a and PR-1b in plant defense response.

Genetic Variations of Trichophyton rubrum Clinical Isolates from Korea

  • Yoon, Nam-Sup;Kim, Hyunjung;Park, Sung-Bae;Park, Min;Kim, Sunghyun;Kim, Young-Kwon
    • Biomedical Science Letters
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    • v.24 no.3
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    • pp.221-229
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    • 2018
  • Trichophyton rubrum is one of the well-known pathogenic fungi and causes dermatophytosis and cutaneous mycosis in human world widely. However, there are not an available sequence type (ST) classification methods and previous studies for T. rubrum until now. Therefore, currently, molecular biological tools using their DNA sequences are used for genotype identification and classification. In the present study, in order to characterize the genetic diversity and the phylogenetic relation of T. rubrum clinical isolates, five different housekeeping genes, such as actin (ACT), calmodulin (CAL), RNA polymerase II (RPB2), superoxide dismutase 2 (SOD2), and ${\beta}$-tubulin (BT2) were analyzed using by multilocus sequence typing (MLST). Also, DNA sequence analysis was performed to examine the differences between the sequences of Trichophyton strains and the identified genetic variations sequence. As a result, most of the sequences were shown to have highly matched rates in their housekeeping genes. However, genetic variations were found on three different positions of ${\beta}$-tubulin gene and were shown to have changed from $C{\rightarrow}G$ (1766), $G{\rightarrow}T$ (1876), and $C{\rightarrow}A$ (1886). To confirm the association with T. rubrum inheritance, a phylogenetic tree analysis was performed. It was classified as four clusters, but there was little significant correlation. Even so, MLST analysis is believed to be helpful for determining the genetic variations of T. rubrum in cases where there is more large-scale data accumulation. In conclusion, the present study demonstrated the first MLST analysis of T. rubrum in Korea and explored the possibility that MLST could be a useful tool for studying the epidemiology and evolution of T. rubrum through further studies.

Characterization of Quintinite Particles in Fluoride Removal from Aqueous Solutions

  • Kim, Jae-Hyun;Park, Jeong-Ann;Kang, Jin-Kyu;Son, Jeong-Woo;Yi, In-Geol;Kim, Song-Bae
    • Environmental Engineering Research
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    • v.19 no.3
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    • pp.247-253
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    • 2014
  • The aim of this study was to characterize quintinite in fluoride removal from aqueous solutions, using batch experiments. Experimental results showed that the maximum adsorption capacity of fluoride to quintinite was 7.71 mg/g. The adsorption of fluoride to quintinite was not changed at pH 5-9, but decreased considerably in highly acidic (pH < 3) and alkaline (pH > 11) solution conditions. Kinetic model analysis showed that among the three models (pseudo-first-order, pseudo-second-order, and Elovich), the pseudo-second-order model was the most suitable for describing the kinetic data. From the nonlinear regression analysis, the pseudo-second-order parameter values were determined to be $q_e=0.18mg/g$ and $k_2=28.80g/mg/hr$. Equilibrium isotherm model analysis demonstrated that among the three models (Langmuir, Freundlich, and Redlich-Peterson), both the Freundlich and Redlich-Peterson models were suitable for describing the equilibrium data. The model analysis superimposed the Redlich-Peterson model fit on the Freundlich fit. The Freundlich model parameter values were determined from the nonlinear regression to be $K_F=0.20L/g$ and 1/n=0.51. This study demonstrated that quintinite could be used as an adsorbent for the removal of fluoride from aqueous solutions.

Transcriptome sequencing revealed the inhibitory mechanism of ketoconazole on clinical Microsporum canis

  • Wang, Mingyang;Zhao, Yan;Cao, Lingfang;Luo, Silong;Ni, Binyan;Zhang, Yi;Chen, Zeliang
    • Journal of Veterinary Science
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    • v.22 no.1
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    • pp.4.1-4.13
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    • 2021
  • Background: Microsporum canis is a zoonotic disease that can cause dermatophytosis in animals and humans. Objectives: In clinical practice, ketoconazole (KTZ) and other imidazole drugs are commonly used to treat M. canis infection, but its molecular mechanism is not completely understood. The antifungal mechanism of KTZ needs to be studied in detail. Methods: In this study, one strain of fungi was isolated from a canine suffering with clinical dermatosis and confirmed as M. canis by morphological observation and sequencing analysis. The clinically isolated M. canis was treated with KTZ and transcriptome sequencing was performed to identify differentially expressed genes in M. canis exposed to KTZ compared with those unexposed thereto. Results: At half-inhibitory concentration (½MIC), compared with the control group, 453 genes were significantly up-regulated and 326 genes were significantly down-regulated (p < 0.05). Quantitative reverse transcription polymerase chain reaction analysis verified the transcriptome results of RNA sequencing. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the 3 pathways of RNA polymerase, steroid biosynthesis, and ribosome biogenesis in eukaryotes are closely related to the antifungal mechanism of KTZ. Conclusions: The results indicated that KTZ may change cell membrane permeability, destroy the cell wall, and inhibit mitosis and transcriptional regulation through CYP51, SQL, ERG6, ATM, ABCB1, SC, KER33, RPA1, and RNP genes in the 3 pathways. This study provides a new theoretical basis for the effective control of M. canis infection and the effect of KTZ on fungi.

Design and heat transfer optimization of a 1 kW free-piston stirling engine for space reactor power system

  • Dai, Zhiwen;Wang, Chenglong;Zhang, Dalin;Tian, Wenxi;Qiu, Suizheng;Su, G.H.
    • Nuclear Engineering and Technology
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    • v.53 no.7
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    • pp.2184-2194
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    • 2021
  • The Free-Piston Stirling engine (FPSE) is of interest for many research in aerospace due to its advantages of long operating life, higher efficiency, and zero maintenance. In this study, a 1-kW FPSE was proposed by analyzing the requirements of Space Reactor Power Systems (SRPS), of which performance was evaluated by developing a code through the Simple Analysis Method. The results of SAM showed that the critical parameters of FPSE could satisfy the designed requirements. The heater of the FPSE was designed with the copper rectangular fins to enhance heat transfer, and the parametric study of the heater was performed with Computational Fluid Dynamics (CFD) software STAR-CCM+. The Performance Evaluation Criteria (PEC) was used to evaluate the heat transfer enhancement of the fins in the heater. The numerical results of the CFD program showed that pressure drop and Nusselt number ratio had a linear growth with the height of fins, and PEC number decreased as the height of fins increased, and the optimum height of the fin was set as 4 mm according to the minimum heat exchange surface area. This paper can provide theoretical supports for the design and numerical analysis of an FPSE for SRPSs.