Detailed Information on Publication Record
2002
The effect of cultivation conditions on protein composition of membrane of Paracoccus denitrificans
BOUCHAL, Pavel and Igor KUČERABasic information
Original name
The effect of cultivation conditions on protein composition of membrane of Paracoccus denitrificans
Name in Czech
Vliv kultivačních podmínek na proteinové složení membránové frakce Paracoccus denitrificans
Authors
BOUCHAL, Pavel (203 Czech Republic, guarantor) and Igor KUČERA (203 Czech Republic)
Edition
Brno, VI. Pracovní setkání biochemiků a molekulárních biologů. Sborník příspěvků. p. 67-67, 2002
Publisher
Masarykova univerzita
Other information
Language
English
Type of outcome
Stať ve sborníku
Field of Study
10600 1.6 Biological sciences
Country of publisher
Czech Republic
Confidentiality degree
není předmětem státního či obchodního tajemství
RIV identification code
RIV/00216224:14310/02:00019686
Organization unit
Faculty of Science
ISBN
80-210-2777-0
Keywords in English
Paracoccus denitrificans; membrane proteins; proteome; 2-D electrophoresis
Změněno: 25/5/2006 12:18, doc. Mgr. Pavel Bouchal, Ph.D.
V originále
New method using two-dimensional gel electrophoresis was optimised for Paracoccus denitrificans cell envelope study. This approach consist of sample preparation especially optimised for Paracoccus denitrificans membrane proteins, carrier-ampholyte isoelectric focusation in the first dimension (CA-IEF) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension. Acquired 2-D gels were compared using image analysis and PDQUEST software. Various conditions of initial solubilisation using various detergents were tested. Sodium dodecyl sulphate (SDS), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfo-nate (CHAPS), dodecyl-â-D-maltoside and 6-aminocaproic acid were used for this purpose. Sample buffer and IEF gel composition effects on 2-D gels resolution were also tested. Extraction using free detergent was compared with extraction using diluted detergent in mixture with sample buffer. Dilution ratio between presolubilised extract and sample buffer was optimised. Focusation algorithm was also optimised for this purpose.
In Czech
Byla optimalizována nová metoda k analýze membránové frakce bakterie Paracoccus denitrificans za použití isoelektrické fokusace s nosnými amfolyty a dodecylsulfátové polyakrylamidové elektroforézy. Při optimalizaci byly testovány různé postupy přípravy vzorku za použití různých detergentů. 2-D gely byly vyhodnocovány pomocí software PDQUEST.
Links
GA203/01/1589, research and development project |
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MSM 143100005, plan (intention) |
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