Comparison of different methods of cell lysis and protein measurements in Clostridium perfringens: application to the cell volume determination.
Four cell lysis methods (NaOH-SDS solubilization, French press treatment, sonication, mutanolysin treatment) and three methods of protein assays (Lowry, Bradford, Pierce) were studied for their applicability to determination of cell volume in Clostridium perfringens NCTC 8798 cell suspensions. Protein contents were higher after a mechanical disruption of the cells than with the other techniques of lysis. The lowest concentrations of protein were obtained with the Bradford procedure. With each of the three protein assay methods, Clostridium perfringens NCTC 8798 protein cell contents were 45% to 58% of protein. Other factors possibly involved in variations of the intracellular volume measurements were examined. A control of the level of protein concentration in the test sample and the type of silicone oil used for the centrifugation were of prime importance during sample preparation. Under our conditions, an intracellular volume of 4 microl/(mg of protein) was routinely found for Clostridium perfringens NCTC 8798.
Guerlava
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Laboratoire de Génie des Procédés et de Technologie Alimentaires, Institut National de la Recherche Agronomique, 369 rue J. Guesde, BP 39, F-59651 Villeneuve d'Ascq Cedex, France.
Izac
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Tholozan
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D001426:Bacterial Proteins / Q000032:analysis* / Q000187:drug effects; D001433:Bacteriolysis; D002250:Carbon Radioisotopes / Q000032:analysis; D002474:Cell-Free System / Q000187:drug effects; D003016:Clostridium perfringens / Q000737:chemistry* / Q000166:cytology* / Q000187:drug effects; D003593:Cytoplasm / Q000737:chemistry / Q000187:drug effects / Q000382:microbiology; D003600:Cytosol / Q000737:chemistry / Q000187:drug effects / Q000382:microbiology; D003911:Dextrans / Q000032:analysis / Q000493:pharmacokinetics; D010450:Endopeptidases / Q000494:pharmacology; D009821:Oils / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D012972:Sodium Hydroxide / Q000494:pharmacology; D012995:Solubility; D013010:Sonication; D013012:Sorbitol / Q000032:analysis / Q000493:pharmacokinetics
D001426:Bacterial Proteins; D002250:Carbon Radioisotopes; D003911:Dextrans; D009821:Oils; D012967:Sodium Dodecyl Sulfate; D013012:Sorbitol; D012972:Sodium Hydroxide; D010450:Endopeptidases; C021816:mutanolysin