A simple and rapid method for the differentiation and identification of thermophilic bacteria

2006 ◽  
Vol 52 (8) ◽  
pp. 753-758 ◽  
Author(s):  
Bin Liu ◽  
Hebin Li ◽  
Suijie Wu ◽  
Xiaobo Zhang ◽  
Lianhui Xie
Keyword(s):  
Hot Spring ◽  
Thermophilic Bacteria ◽  
Random Amplified Polymorphic Dna ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cell Protein ◽  
Protein Patterns ◽  
Simple Method ◽  
Whole Cell ◽  
Sds Page

In total, 170 strains of thermophilic bacteria were isolated from deep-sea hydrothermal fields in the Pacific Ocean and a hot spring in Xiamen of China. To facilitate the identification of thermophilic strains, sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS–PAGE) of whole-cell proteins of these strains was first performed. The results showed that there exist four different protein patterns, indicating that the 170 strains might belong to four species or genera. The RAPD (random amplified polymorphic DNA) profiles of nine representative strains were consistent with those of SDS–PAGE. To further identify the species of the nine strains, their 16S rDNA sequences were analyzed. The results showed that the nine strains fell into four species of three genera, which was the same as revealed by SDS–PAGE. Therefore, SDS–PAGE of whole-cell proteins could be used as a rapid and simple method for the discrimination of thermophilic bacteria as the first step of species identification.Key words: thermophilic bacteria, SDS–PAGE, whole-cell protein, discrimination.

scholarly journals  Analysis of whole cell protein profiles of Salmonella serovars isolated from chicken, turkey and sheep faeces by SDS-PAGE

2010 ◽  
Vol 55 (No. 6) ◽  
pp. 259-263 ◽  
Author(s):  
A. Aksakal
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Visual Assessment ◽  
Cell Protein ◽  
Protein Profiles ◽  
Whole Cell ◽  
Sds Page ◽  
Molecular Masses ◽  
Whole Cell Protein ◽  
Salmonella Serovars

This study was carried out to determine the whole cell protein profiles of Salmonella serovars from chicken, turkey and sheep faeces by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A total of 34 Salmonella strains were included in the study, 14 of them were isolated from chicken, 14 from turkey and six from sheep. SDS-PAGE was carried out using 12% (w/v) separating and 4% (w/v) stacking gels. The results showed more than 30 protein bands ranging in size from 97 kDa (kilodaltons) to below 14.4 kDa as determined by visual assessment of their approximate molecular masses. Protein bands of 78.1, 51.2, 41.5, 37.3, 35.1, 33.9, 30.7, 27.6, 25.4, and 24 kDa were detected in all Salmonella serovars. Salmonella strains used in this study were closely related and could not be differentiated depending on the whole cell protein profiles using SDS-PAGE.

scholarly journals Electrophoretic Identification of Garlic and Garlic Products

1996 ◽  
Vol 79 (6) ◽  
pp. 1466-1470 ◽  
Author(s):  
Emiko Mochizuki ◽  
Takao Yamamoto ◽  
Sumiko Suzuki ◽  
Hiroyuki Nakazawa
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Silver Staining ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Garlic Extract ◽  
Constant Current ◽  
Protein Patterns ◽  
Simple Method ◽  
Sds Page

Abstract We developed a rapid and simple method for identifying garlic and garlic products using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) with silver staining. Samples were homogenized with 1 % SDS or 6M urea and centrifuged. Supernatant containing garlic proteins was mixed with the same volume of loading buffer containing SDS and mercaptoethanol, heated in boiling water for 2 min, and applied to the wells of a ready-to-use polyacrylamide gradient gel (4–20%). Electrophoresis was performed 20 mA constant current for 2 h. The gel was stained with a silver staining kit and dried. Protein patterns of garlic and garlic products are different from those of other Allium plants such as onion, rakkyo, and caucas. The method was used to analyze samples of spice and garlic clove products. Absence of protein bands in garlic extract products suggests the products may contain less proteins and/or denatured proteins.

scholarly journals Stenotrophomonas africana Drancourt et al. 1997 is a later synonym of Stenotrophomonas maltophilia (Hugh 1981) Palleroni and Bradbury 1993

2004 ◽  
Vol 54 (4) ◽  
pp. 1235-1237 ◽  
Author(s):  
Tom Coenye ◽  
Elke Vanlaere ◽  
Enevold Falsen ◽  
Peter Vandamme
Keyword(s):  
Dna Hybridization ◽  
Stenotrophomonas Maltophilia ◽  
Biochemical Characterization ◽  
Cell Protein ◽  
Protein Patterns ◽  
Whole Cell ◽  
Sds Page ◽  
Binding Level ◽  
Reference Strains ◽  
Type Strains

Type and reference strains of Stenotrophomonas maltophilia and Stenotrophomonas africana were compared with each other and with the type strains of other Stenotrophomonas species, using SDS-PAGE of whole-cell proteins, DNA–DNA hybridization and extensive biochemical characterization. S. maltophilia LMG 958T and S. africana LMG 22072T had very similar whole-cell-protein patterns and were also biochemically very similar. A DNA–DNA binding level of 70 % between both type strains confirmed that S. africana and S. maltophilia represent the same taxon. It is concluded that S. africana is a later synonym of S. maltophilia.

Protein Expression in Vibrio parahaemolyticus 690 Subjected to Sublethal Heat and Ethanol Shock Treatments

2008 ◽  
Vol 71 (11) ◽  
pp. 2289-2294 ◽  
Author(s):  
MING-LUN CHIANG ◽  
WEI-LI HO ◽  
ROCH-CHUI YU ◽  
CHENG-CHUN CHOU
Keyword(s):  
Heat Shock ◽  
Vibrio Parahaemolyticus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Test Organism ◽  
Protein Patterns ◽  
One Dimensional ◽  
Sds Page ◽  
Molecular Masses ◽  
Stressed Cells

Cells of Vibrio parahaemolyticus 690 were subjected either to heat shock at 42°C for 45 min or to ethanol shock in the presence of 5% ethanol for 60 min. The protein profiles of the unstressed and stressed V. parahaemolyticus cells were compared. Additionally, the induction of DnaK- and GroEL-like proteins in the unstressed and stressed cells of V. parahaemolyticus was also examined. Analysis with one-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) indicated that three proteins with molecular masses of 93, 77, and 58 kDa were induced by both heat shock and ethanol shock. The protein patterns revealed by two-dimensional electrophoresis were more detailed than those revealed by one-dimensional SDS-PAGE. It was found that heat shock and ethanol shock affected the expression of a total of 28 proteins. Among them, four proteins with molecular masses of 94, 32.1, 26.7, and 25.7 kDa were enhanced by both heat shock and ethanol shock. Furthermore, immunoblot analysis showed the presence of a GroEL-like protein with a molecular mass of 61 kDa in the test organism, with the heat-shocked and ethanol-shocked cells producing a GroEL-like protein in a larger quantity than the unstressed cells. However, DnaK-like protein was not detectable in either the unstressed or the stressed cells.

Whole-cell protein and ITS rDNA profiles as diagnostic tools to discriminate Fusarium avenaceum intraspecific variability and associated virulence

2009 ◽  
Vol 55 (2) ◽  
pp. 117-125 ◽  
Author(s):  
V. Vujanovic ◽  
S. Vidovic ◽  
M. R. Fernandez ◽  
P. Daida ◽  
D. Korber
Keyword(s):  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Intraspecific Variability ◽  
Fusarium Avenaceum ◽  
Cell Protein ◽  
Diagnostic Tools ◽  
Group Method ◽  
Head Blight ◽  
Sds Page

A total of 91 isolates of Fusarium avenaceum were regrouped into 15 phenotypes and 10 vegetative compatibility groups showing specific one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (1-D SDS–PAGE) protein profiles and less-specific internal transcribed spacer rDNA profiles. Each isolate possessed reproducible signature protein bands. Indeed, the unweighted pair group method with arithmetic averages clustering revealed that the protein profile of each group of isolates correlated with fungus virulence. The use of SDS–PAGE offers a simple and sensitive technique for routine differentiation between pathogenic and nonpathogenic isolates within unknown F. avenaceum populations. The discovery has significant implications for risk assessment of cereal yield to ensure food and feed safety. This low-cost approach has the potential to be optimized and extended to a broad spectrum of Fusarium head blight pathogens.

scholarly journals Specificity of Rabbit Antisera against Lipopolysaccharide of Acinetobacter

1998 ◽  
Vol 36 (5) ◽  
pp. 1245-1250 ◽  
Author(s):  
Ralph Pantophlet ◽  
Lore Brade ◽  
Lenie Dijkshoorn ◽  
Helmut Brade
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Proteinase K ◽  
Accurate Identification ◽  
Simple Method ◽  
Whole Cell ◽  
Cell Lysates ◽  
O Antigen ◽  
Homologous Antigen ◽  
Routine Identification

Acinetobacter has been reported to be involved in hospital-acquired infections with increasing frequency. However, clinical laboratories still lack simple methods that allow the accurate identification of Acinetobacter strains at the species level. For this study, proteinase K-digested whole-cell lysates from 44 clinical and environmental isolates were investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with hyperimmune rabbit sera to examine the possibility of developing a serotyping scheme based on the O antigen of Acinetobacterlipopolysaccharide (LPS). The antisera, obtained by immunization of rabbits with 13 of the heat-killed isolates investigated, were characterized by Western blotting and enzyme immunoassay by using proteinase K-digested whole-cell lysates and phenol-water-extracted LPS as antigens. In both assays, the antisera were shown to be highly specific for the homologous antigen. In addition, assignment ofAcinetobacter LPS to the smooth or the rough phenotype was shown not to be reliable when it was based only on the results obtained with silver-stained gels. O-antigen reactivity, determined by Western blot analysis, was observed with 11 of the 31 isolates, most of which belonged to the species Acinetobacter baumannii (DNA group 2) and the unnamed DNA group 3. Interestingly, some O antigens were found in a DNA group different from that of the strain used for immunization. The results indicate that O serotyping ofAcinetobacter strains is feasible and thus may provide a simple method for the routine identification of these opportunistic pathogens.

scholarly journals Comparative studies of Yersinia pestis outer membrane isolation techniques and their potential use in plague epidemiology

1990 ◽  
Vol 32 (2) ◽  
pp. 78-83 ◽  
Author(s):  
Frederico Guilherme Coutinho Abath ◽  
Luís Carlos de Sousa Ferreira
Keyword(s):  
Yersinia Pestis ◽  
Outer Membrane ◽  
Outer Membrane Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Patterns ◽  
Isolation Techniques ◽  
Sds Page ◽  
Triton X ◽  
Potential Use

In the present study three techniques for obtaining outer membrane enriched fractions from Yersinia pestis were evaluated. The techniques analysed were: differential solubilization of the cytoplasmic membrane with Sarkosyl or Triton X-100, and centrifugation in sucrose density gradients. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of outer membrane isolated by the different methods resulted in similar protein patterns. The measurement of NADH-dehydrogenase and succinate dehydrogenase (inner membrane enzymes) indicated that the outer membrane preparations obtained by the three methods were pure enough for analytical studies. In addition, preliminary evidences on the potential use of outer membrane proteins for the identification of geographic variants of Y. pestis wild isolates are presented.

scholarly journals Evaluation of numerical analysis of SDS-PAGE of protein patterns for typingEnterobacter cloacae

1989 ◽  
Vol 103 (2) ◽  
pp. 265-274 ◽  
Author(s):  
M. Costas ◽  
L. L. Sloss ◽  
R. J. Owen ◽  
M. A. Gaston
Keyword(s):  
Numerical Analysis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Phage Typing ◽  
Clinical Isolates ◽  
Protein Patterns ◽  
One Dimensional ◽  
Sds Page ◽  
Typing Methods ◽  
Type Strains

SUMMARYTwenty cultures comprising 13 clinical isolates ofEnterobacter cloacaefrom two hospitals. the type and another reference stain ofE. cloacaeand the type strains of four otherEnterobactersp. and ofEscherichia coli, were characterized by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDSPAGE) of whole-cell proteins. The protein patterns were highly reproducible and were used as the basis of a numerical analysis which divided the clinical isolates into nine clearly defined protein types. Comparison with established typing methods indicated that the discrimination of SDS-PAGE was similar to that achieved with conventional typing methods and all strain groups recognized by combined sero/phage typing were also found by SDS-PAGE. In addition, protein typing sub-divided a group of four serotype O3 isolates that were difficult to distinguish by phage typing. We conclude that high-resolution SDS-PAGE of proteins provides an effective method of typing isolates ofE. cloacae.

scholarly journals Subtyping ofHaemophilus influenzaetype b strains from Europe and North America by SDS—PAGE of whole-cell polypeptides: the geographical distribution of subtypes

1989 ◽  
Vol 102 (1) ◽  
pp. 11-19 ◽  
Author(s):  
T. H. Pennington ◽  
E. M. Freebairn
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Invasive Disease ◽  
Wide Distribution ◽  
Dice Coefficient ◽  
Whole Cell ◽  
Geographical Variations ◽  
Sds Page ◽  
The Usa ◽  
The Common

SUMMARYOne hundred and nine strains ofHaemophilus influenzaetype b were subtyped by sodium dodecyl sulphate polyacrylamide gel electrophoresis of whole-cell polypeptides. Twenty-one strains from England, 44 from Scotland, 8 from Sweden, 6 from the Netherlands and 30 from the USA were examined. Some of these strains had been subtyped by outer membrane protein analysis; most of the strains had been isolated from cases of invasive disease.Comparison of polypeptide profiles using the Dice coefficient of similarity showed that the majority of European strains were closely related and formed a single large group. Four smaller groups were identified; three of these included American and European strains, indicating a world-wide distribution of subtypes. However, the common European and American subtypes fell into different groups, indicating the existence of marked geographical variations in subtype frequency.

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