• Title/Summary/Keyword: Two-Dimensional Gel Electrophoresis

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Analysis of Differential-expressed Proteins of Acidithiobacillus ferrooxidans Grown under Phosphate Starvation

  • He, Zhiguo;Zhong, Hui;Hu, Yuehua;Xiao, Shengmu;Liu, Jiarshe;Xu, Jin;Li, Guiyuen
    • BMB Reports
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    • v.38 no.5
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    • pp.545-549
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    • 2005
  • Acidithiobacillus ferrooxidans is one of the most important bacterium used in bioleaching, and can utilize $Fe^{2+}$ or sulphide as energy source. Growth curves for Acidithiobacillus ferrooxidans under phosphate starvation and normal condition have been tested, showing lag, logarithmic, stationary and aging phases as seen in other bacteria. The logarithmic phases were from 10 to 32 hours for Acidithiobacillus ferrooxidans cultivated with normal cultivating condition and from 20 to 60 hrs for Acidithiobacillus ferrooxidans cultivated phosphate starvation. Differences of protein patterns of Acidithiobacillus ferrooxidans growing in case of normal or phosphate starvation were separately investigated after cultivation at $30^{\circ}C$ by the analysis of two-dimensional gel electrophoresis (2-DE), matrix-assisted laser desorption/ionization (MALDI)-Mass spectrometry. There were total 6 protein spots identified, which were Recombination protein recA, RNA helicase, AP2 domain-containing transcription factor, NADH dehydrogenase I chain D, Hyothetical protein PF1669, and Transaldolase STY3758. From the 6 identified protein spots, 3 proteins were found to be decreased in expression at the cultivating condition of phosphate starvation, while another three upregulated.

Proteomic Analysis of Polycyclic Aromatic Hydrocarbons (PAHs) Degradation and Detoxification in Sphingobium chungbukense DJ77

  • Lee, Soo Youn;Sekhon, Simranjeet Singh;Ban, Yeon-Hee;Ahn, Ji-Young;Ko, Jung Ho;Lee, Lyon;Kim, Sang Yong;Kim, Young-Chang;Kim, Yang-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1943-1950
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    • 2016
  • Polycyclic aromatic hydrocarbons (PAHs) are commonly present xenobiotics in natural and contaminated soils. We studied three (phenanthrene, naphthalene, and biphenyl) xenobiotics, catabolism, and associated proteins in Sphingobium chungbukense DJ77 by two-dimensional gel electrophoresis (2-DE) analysis. Comparative analysis of the growth-dependent 2-DE results revealed that the intensity of 10 protein spots changed identically upon exposure to the three xenobiotics. Among the upregulated proteins, five protein spots, which were putative dehydrogenase, dioxygenase, and hydrolase and involved in the catabolic pathway of xenobiotic degradation, were induced. Identification of these major multifunctional proteins allowed us to map the multiple catabolic pathway for phenanthrene, naphthalene, and biphenyl degradation. A part of the initial diverse catabolism was converged into the catechol degradation branch. Detection of intermediates from 2,3-dihydroxy-biphenyl degradation to pyruvate and acetyl-CoA production by LC/MS analysis showed that ring-cleavage products of PAHs entered the tricarboxylic acid cycle, and were mineralized in S. chungbukense DJ77. These results suggest that S. chungbukense DJ77 completely degrades a broad range of PAHs via a multiple catabolic pathway.

Cancer Diagnosis System using Genetic Algorithm and Multi-boosting Classifier (Genetic Algorithm과 다중부스팅 Classifier를 이용한 암진단 시스템)

  • Ohn, Syng-Yup;Chi, Seung-Do
    • Journal of the Korea Society for Simulation
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    • v.20 no.2
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    • pp.77-85
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    • 2011
  • It is believed that the anomalies or diseases of human organs are identified by the analysis of the patterns. This paper proposes a new classification technique for the identification of cancer disease using the proteome patterns obtained from two-dimensional polyacrylamide gel electrophoresis(2-D PAGE). In the new classification method, three different classification methods such as support vector machine(SVM), multi-layer perceptron(MLP) and k-nearest neighbor(k-NN) are extended by multi-boosting method in an array of subclassifiers and the results of each subclassifier are merged by ensemble method. Genetic algorithm was applied to obtain optimal feature set in each subclassifier. We applied our method to empirical data set from cancer research and the method showed the better accuracy and more stable performance than single classifier.

Preliminary Proteomic Analysis of Thiobacillus ferrooxidans Growing on Elemental Sulphur and Fe2+ Separately

  • He, Zhi-guo;Hu, Yue-Hua;Zhong, Hui;Hu, Wei-Xin;Xu, Jin
    • BMB Reports
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    • v.38 no.3
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    • pp.307-313
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    • 2005
  • Thiobacillus ferrooxidans is one of the most important bacterium used in bioleaching, and can utilize $Fe^{2+}$ or sulphide as energy source. Growth curves for Thiobacillus ferrooxidans have been tested, which show lag, logarithmic, stationary and aging phases as seen in other bacteria. The logarithmic phases were from 10 to 32 hours for Thiobacillus ferrooxidans cultivated with $Fe^{2+}$ and from 4 to 12 days for Thiobacillus ferrooxidans cultivated with elemental sulphur. Differences of protein patterns of Thiobacillus ferrooxidans growing on elemental sulphur and $Fe^{2+}$ separately were investigated after cultivation at $30^{\circ}C$ by the analysis of two-dimensional gel electrophoresis (2-DE), matrix-assisted laser desorption/ ionization (MALDI)-Mass spectrometry and ESI-MS/MS. From the 7 identified protein spots, 11 spots were found more abundant when growing on elemental sulphur. By contrast 6 protein spots were found decreased at elemental cultivation condition. Among the proteins identified, cytochrome C have been previously identified as necessary elements of electron-transfering pathway for Thiobacillus ferrooxidans to oxidize $Fe^{2+}$; ATP synthase alpha chain and beta are expressed increased when Thiobacillus ferrooxidans cultivated with $Fe^{2+}$ as energy source. ATP synthase Beta chain is the catalytic subunit, and ATP synthase alpha chain is a regulatory subunit. The function of ATPase produces ATP from ADP in the presence of a proton gradient across the membrane.

Intramolecular DNA Triplexes in Escherichia. coli (Methylase를 사용한 Escherichia coli에서 Triplex 존재에 관한 연구)

  • Rhim, Hyangshuk;Kim, Sungjo;Kang, Seongman
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.231-235
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    • 1998
  • We have introduced a genetic assay to study the existence of intramolecular triplexes in Escherichia coli. A plasmid containing the gene that encodes a temperature-sensitive EcoRI methylase was cotransformed with different plasmids containing inserts, $(G)_9AATTC(G)_9$ and $(GAA)_9TTC(GAA)_8$, that are able to form intramolecular triplexes in vitro. Inhibition of methylation in vivo was found for $(G)_9AATTC(G)_9$ and $(GAA)_9TTC(GAA)_8$, suggesting that the pur pyr sequences adopt unusual strucures in E. coli. In addition, experiments using two dimensional gel electrophoresis confirmed that intramolecular triplexes are formed for the pur pyr sequences under negative supercoiling. These results demonstrate the existence of intramolecular triplexes in E. coli.

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Comparisons Among the Fishes of Genus Liobagrus in Korea by Their Morphology and Electrophoretic Patterns of Proteins (形態 및 蛋白質 電氣泳動像에 依한 韓國産 퉁가리屬 魚類의 比較)

  • 손영목;최의열;안태인
    • The Korean Journal of Zoology
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    • v.27 no.1
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    • pp.25-34
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    • 1984
  • Two species of fishes of Genus Liobagrus that had been collected from three separate river systems in Korea were compared by their morphology and gel electrophoretic patterns of water soluble proteins. In morphology L. andersoni from Han River was distinctly different from L. mediadiposalis collected from both Gum River and Nagdong River. But L. andersoni collected from Gum River not only showed partial similarity for both groups of fishes above in their classifying characteristics but also had a unique ratio of body width/standard length. Comparable similarity and difference among the three groups were also noted in their protein patterns of SDS polyacrylamide gel electrophoresis of various tissues. The difference was clearer and more distinct in the gel run with muscle proteins. A couple of more distinctly different low molecular polypeptides were detected by two dimensional gel electrophoresis. Since the protein patterns shown in this study are not only agreeable with the morphological results but also provide detailed comparisons, fishes of Liobagrus from various water sources can be classified reliably by gel electrophoresis. On the bases of tghe findings above, L. andersoni like fishes collected from Gum River should no longer be classified as L. andersoni. Before naming them as an independent species, the possibility of natural hybrid between the two defined species, or of a varient of L. andersoni by geograpic isolation should be tested.

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Differentiation of Three Lactobacillus rhamnosus Strains (E/N, Oxy, and Pen) by SDS-PAGE and Two-Dimensional Electrophoresis of Surface-Associated Proteins

  • Jarocki, P.;Podlesny, M.;Wasko, A.;Siuda, A.;Targonski, Z.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.558-562
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    • 2010
  • SDS-PAGE of extracted surface-associated proteins of Lactobacillus rhamnosus strains E/N, Oxy, and Pen, was performed. The obtained protein patterns allowed differentiation of the examined strains, which was not accomplished by the commonly used RAPD genotypic method. The differentiation by the SDS-PAGE method proved to be a useful tool for strain-specific identification, which was further confirmed by 2DE analysis. Therefore, it can be used as an alternative or complementary method for both conventional and genotypic identification procedures, especially when closely related lactobacilli isolates are identified.

Dimerization of Thyroglobulin in the Endoplasmic Reticulum (Thyroglobulin의 소포체내 dimerization)

  • 권오유;신기선
    • Journal of Life Science
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    • v.13 no.3
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    • pp.248-251
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    • 2003
  • The kinetics of dimerization of a newly synthesized thyroglobulin (Tg), the precursor protein in the manufacture of thyroid hormone, was investigated in the endoplasmic reticulum of thyrocytes FRTL-5 cell line. The folded monomeric Tg was first detectable in a conformationally unstable form, from the examination of lysates of pulse labeled cultured thyrocytes by denaturing and nondenaturing gel electrophoresis by 15 min after biosynthesis. The first dimeric Tg was formed by 30 min after; the monomer declined and the dimer progressively increased, and 40 min after remarkable dimeric Tg form was found. Finally, dimerization was complete at 60 min after.

Profile Analysis of Proteins Related with Hydrogen Peroxide Response in Strep-tomyces coelicolor (Muller) (Streptomyces coelicolor (Muller)의 과산화수소 대응 반응에 관련된 단백질 양상의 분석)

  • 정혜정;노정혜
    • Korean Journal of Microbiology
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    • v.31 no.2
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    • pp.166-174
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    • 1993
  • Streptomyces coeUc%r (Muller) cells were treated with $100 \mu$M hydrogen peroxide for I hour and proteins synthesized during hydrogen peroxide stress were labeled with L-[$^{35}S$]-methionine. Total cellular proteins were extracted and analyzed by two-dimensional polyacrylamide gel electrophoresis. In exponential growth phase, synthesis of about 100 proteins was increased by hydrogen peroxide treatment. These proteins were named as Pin (£eroxide-inducib]e) proteins and classified into 4 subgroups according to their induction time after hydrogen peroxide treatment. About 60 of them were found to be induced within 20 minutes and maintained throughout I hour of treatment. In stationary growth phase. synthesis of 62 proteins was increased by hydrogen peroxide and 21 of them were the same Pins found in exponential growth phase. Proteins from the mutants which are resistant to hydrogen peroxide were obtained in exponential growth phase and compared with those from the wild type on two-dimensional gel. The three mutants, N7, N9. and N24, were found to have higher constitutive leve]s of ]5, 17, and 15 Pin proteins respectively, than the wild type. 9 of these Pin proteins (D74.7a, E76.0c, E23.3. F50.7, F47.2a. F25.5, G39.6b, G24.0, H39.6a) increased in two of the three mutants and 3 proteins (F39.7, H6I.7. 120.8) increased in all of the three mutants. These proteins might play important roles in the response of S. coelic%r to hydrogen peroxide.

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Cloned Placenta of Korean Native Calves Died Suddenly at Two Months after Birth Displays Differential Protein Expression

  • Kim Hong Rye;Kang Jae Ku;Lee Hye Ran;Yoon Jong Taek;Seong Hwan Hoo;Jung Jin Kwan;Park Chang Sik;Jin Dong Il
    • Reproductive and Developmental Biology
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    • v.29 no.2
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    • pp.63-68
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    • 2005
  • Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, $H^+-transporting$ ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.