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  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.

A rapid method of species identification of wild chironomids (Diptera: Chironomidae) via electrophoresis of hemoglobin proteins in sodium dodecyl sulfate polyacrylamide gel (SDS–PAGE)

2014 ◽  
Vol 104 (5) ◽  
pp. 639-651 ◽  
Author(s):  
J.T. Oh ◽  
J.H. Epler ◽  
C.S. Bentivegna
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Head Capsule ◽  
Species Level ◽  
Polyacrylamide Gel ◽  
Taxonomic Identification ◽  
Sds Page ◽  
Dodecyl Sulfate

AbstractStudying aquatic benthic macroinvertebrates (BMIs) in the field requires accurate taxonomic identification, which can be difficult and time consuming. Conventionally, head capsule morphology has been used to identify wild larvae of Chironomidae. However, due to the number of species and possible damage and/or deformity of their head capsules, another supporting approach for identification is needed. Here, we provide hemoglobin (Hb) protein in hemolymph of chironomids as a new biomarker that may help resolve some of the ambiguities and difficulties encountered during taxonomic identification. Chironomids collected from two locations in Maine and New Jersey, USA were identified to the genus level and in some cases to the species-level using head capsule and body morphologies. The head capsule for a particular individual was then associated with a corresponding Hb protein profile generated from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE). Distinct Hb profiles were observed from one group (Thienemannimyia) and four genera (Chironomus, Cricotopus, Dicrotendipes, and Glyptotendipes) of chironomids. Several species were polymorphic, having more than one Hb profile and/or having bands of the same size as those of other species. However, major bands and the combination of bands could distinguish individuals at the genus and sometimes species-level. Overall, this study showed that Hb profiles can be used in combination with head capsule morphology to identify wild chironomids.

scholarly journals Protein Profile Study of Some Nigerian Sesame (Sesamum indicum L.) Accessions

2015 ◽  
Vol 3 (2) ◽  
pp. 322-329 ◽  
Author(s):  
Gbenga Olorunshola Alege
Keyword(s):  
Genetic Diversity ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sesamum Indicum ◽  
Genomic Changes ◽  
Sds Page ◽  
Sesame Genotypes ◽  
Species Specific ◽  
Total Seed

This study was carried out to investigate the genetic diversity among 23 sesame (Sesamum indicum L.) accessions obtained from different agro-ecological localities from 10 different states across 4 geopolitical zones in Nigeria using evidence from Sodium Dodecyl Polyacrylamide Gel Electrophoresis (SDS-PAGE). Total seed protein of the studied plants resolved on 12% SDS-PAGE showed variations in numbers and intensity of bands among the different sesame accessions. Thirteen (13) major bands were recorded in this study. Lack of unique band and presence of common band (band 7) among the 23 studied sesame accessions indicate some levels of genetic affinity and evidence of common evolutionary origin of the sesame genotypes. This band can therefore be tagged as species specific band for discriminating Sesamum indicum. Cluster analysis grouped the 23 sesame genotypes into two clusters with similarity coefficient ranging from 0.42 to 0.96 which indicates existence of genetic diversity; therefore there is ample opportunity for improving the 23 sesame genotypes. Variations in protein bands observed among the 23 studied plants could be attributed to genomic changes taken place during species diversification. It can be concluded that genetic diversity existed among Nigerian sesame for the improvement of characters of interest. Accessions 9 (YOL), 15(OTT), 22 (OFF) and 23 (JAL) are therefore recommended for used in future breeding programs for the development of improved sesame varieties.Int J Appl Sci Biotechnol, Vol 3(2): 322-329 DOI: http://dx.doi.org/10.3126/ijasbt.v3i2.12734

scholarly journals Analysis of Whole Cell Protein Profiles by SDS-PAGE to Identify Indigenous Cellulose-producer Acetic Acid Bacteria

2017 ◽  
Vol 21 (2) ◽  
pp. 86
Author(s):  
Sarkono Sarkono ◽  
Soekarti Moeljopawiro ◽  
Bambang Setiaji ◽  
Langkah Sembiring
Keyword(s):  
Acetic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Acetic Acid Bacteria ◽  
Cellular Protein ◽  
Cell Protein ◽  
Phenotypic Traits ◽  
Protein Profiles ◽  
Bacterial Isolates ◽  
Sds Page

This study was carried out to analyze the suitability of the identification of four indigenous cellulose-producing acetic acid bacterial isolates (ANG29, KRE65, ANG32 and SAL53) based on the analysis of whole cellular protein profiles against identification based on phenotypic traits. Whole cellular protein profiles were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method. The whole cellular protein profiles obtained from sample isolates, were compared with reference isolates for species identification. The results showed that based on visual observations can be determined as much as 12 bands of protein with a molecular weight of 19,099 KDa up to 132.182 KDa. Based on the analysis of protein bands were detected visually, fourth indigenous cellulose- producing acetic acid bacterial isolates in the study had a higher similarity profile to the reference strain Gluconacetobacter xylinus BTCC 769 compared with other reference strains namely G. hansenii NBRC 14820T. This condition is consistent with the results of the identification of fourth cellulose producing acetic acid bacterial isolates based on phenotypic traits. Thus, the whole cellular protein profiles by SDS-PAGE technique can be used as a one of method to identification of cellulose producing acetic acid bacterial isolates.

scholarly journals Protein Profiles from Intact Cells as a Tool in Bifidobacterium Characteristics

2012 ◽  
Vol 61 (4) ◽  
pp. 305-310 ◽  
Author(s):  
ADAM WAŚKO ◽  
MAGDALENA POLAK-BERECKA ◽  
MICHAŁ KALITA
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Molecular Techniques ◽  
Group Method ◽  
Intact Cells ◽  
Arithmetic Average ◽  
Pair Group ◽  
Sds Page ◽  
Liquid Chromatography Tandem Mass ◽  
Electrophoresis Separation

In this study sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) profiles were analysed and differences were confirmed by a unweighted pair group method with arithmetic average (UPGMA) analysis between bifidobacterial species, such as B. infanis ATCC1567, B. bifidum Bb-12, B. longum KN29, B. catenulatum KD14, and B. animalis BI30. Two dimensional electrophoresis separation profiles were compared, and the most characteristic spots were characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS). We propose proteins extracted from intact cells as an additional trait for bifidobacteria characterization, together with molecular techniques, which can be used to analyze bacterial protein polymorphism and to distinguish among species.

scholarly journals Protein-Based Genetic Diversity Assessment of Tilapia guineensis and Sarotherodon melanotheron Populations from South-West Nigerian Coastal Waters

2020 ◽  
pp. 9-15
Author(s):  
E. A. Ukenye ◽  
I. Megbowon ◽  
M. M. A. Akinwale ◽  
M. A. Fowora ◽  
I. Chidume ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Genetic Variation ◽  
Genetic Similarity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Group Method ◽  
Cichlid Species ◽  
Diversity Assessment ◽  
Sds Page ◽  
Sarotherodon Melanotheron

The present study was carried out to investigate the genetic differences in the Protein banding pattern of Tilapia guineensis and Sarotherodon melanotheron populations in Southwest Nigeria using Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Four populations of Tilapia guineensis and three populations of Sarotherodon melanotheron from Ondo and Lagos states were considered for the study. The sarcoplasmic protein of the studied Cichlid species resolved on 12% SDS-PAGE revealed variations in their genetic diversity indices (number of alleles, shanon information index, heterozygosity and percentage polymorphism). T. guineensis had more proteins and higher genetic diversity as was revealed by the genetic diversity parameters and was found to be more polymorphic with a percentage polymorphism of 78.57% than S. melanotheron (57.14%). The two species had similarity coefficient of 0.82 indicating high genetic similarity between them. UPGMA (Unweighted Pair Group Method with Arithmetic Mean) dendrogram also revealed some level of genetic similarity between the studied populations and among the two species. Analysis of molecular variance (AMOVA) confirmed the low genetic variation among the populations of the cichlid species and demonstrated that genetic variation was mostly within populations in both species. It is established from the study that Tilapia guineensis had higher genetic diversity than Sarotherodon melanotheron and the two species are closely related. Further study involving molecular markers is encouraged to complement this finding.

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 Los patrones electroforéticos de proteínas salivales permiten diferenciar los grupos transandino y cisandino de las especies de Rhodnius de Colombia

Biomédica ◽  
2020 ◽  
Vol 40 (2) ◽  
pp. 404-411
Author(s):  
Arlid Meneses ◽  
Cristian Camilo Rodríguez ◽  
Yazmín Suárez ◽  
Julio César Carranza ◽  
Gustavo Adolfo Vallejo
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Trypanosoma Cruzi ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Group Method ◽  
Pair Group ◽  
Sds Page ◽  
Los Grupos ◽  
Unweighted Pair Group Method

Introducción. Las especies Rhodnius (Hemiptera: Reduviidae: Triatominae) están conformadas por insectos hematófagos vectores de Trypanosoma cruzi, agente etiológico de la enfermedad de Chagas, y T. rangeli, parásito infectivo pero no patógeno para el vertebrado. El estudio de la diversidad proteica de la saliva de estos insectos permite la obtención de perfiles electroforéticos unidimensionales característicos de algunas especies de triatominos. Sin embargo, el reporte de los patrones electroforéticos de proteínas salivales de las especies de Rhodnius ha sido escaso.Objetivo. Hacer un análisis comparativo de los perfiles electroforéticos unidimensionales de las proteínas salivales de R. colombiensis, R. pallescens, R. pictipes, R. prolixus y R. robustus.Materiales y métodos. Se obtuvieron los perfiles electroforéticos de la saliva de las especies en estudio mediante electroforesis en gel de poliacrilamida con dodecilsulfatosódico (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis, SDS-PAGE) y se construyó un fenograma mediante el método UPGMA (Unweighted Pair Group Method Using Arithmetic Averages).Resultados. Los perfiles electroforéticos de las proteínas solubles de saliva presentaron bandas en un rango de masa aproximado de 15 a 45 kDa, los cuales permitieron diferenciar las cinco especies estudiadas. El fenograma reveló la existencia de dos grupos principales: uno conformado por los grupos cisandinos Pictipes y Prolixus y otro constituido por el grupo transandino Pallescens.Conclusiones. Existen diferencias en los perfiles electroforéticos de las proteínas salivales entre R. colombiensis, R. pallescens, R. pictipes, R. prolixus y R. robustus, cuya variabilidad permitió construir un fenograma congruente con los grupos del género Rhodnius.

Phenotypic and genetic diversity of enterococci isolated from Italian cheeses

2001 ◽  
Vol 68 (2) ◽  
pp. 303-316 ◽  
Author(s):  
CHRISTIAN ANDRIGHETTO ◽  
EDO KNIJFF ◽  
ANGIOLELLA LOMBARDI ◽  
SANDRA TORRIANI ◽  
MARC VANCANNEYT ◽  
...  
Keyword(s):  
Species Recognition ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Reference Method ◽  
Antagonistic Activity ◽  
Cell Protein ◽  
Rapd Pcr ◽  
Sds Page ◽  
Carbohydrate Fermentation ◽  
Proteolytic Activities

In the present study, 124 enterococcal strains, isolated from traditional Italian cow, goat and buffalo cheeses, were characterized using phenotypic features and randomly amplified polymorphic DNA polymerase chain reaction (RAPD–PCR). The RAPD–PCR profiles obtained with four primers and five different amplification conditions were compared by numerical analysis and allowed an inter- and intraspecific differentiation of the isolates. Whole-cell protein analysis by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) was used as a reference method for species identification. The strains were identified as Enterococcus faecalis (82 strains), E. faecium (27 strains), E. durans (nine strains), E. gallinarum (four strains) and E. hirae (two strains). Species recognition by means of RAPD–PCR was in agreement with the SDS–PAGE results except for eight strains of E. faecium that clustered in separated groups. On the other hand, phenotypic identification based on carbohydrate fermentation profiles, using the rapid ID 32 STREP galleries, gave different results from SDS–PAGE in 12·1% of the cases. The majority of the strains had weak acidifying and proteolytic activities in milk. One E. faecium strain showed vanA (vancomycin resistance) genotype while four strains showed a β-haemolytic reaction on human blood. Several strains showed antagonistic activity towards indicator strains of Listeria innocua, Clostridium tyrobutyricum and Propionibacterium freudenreichii subsp. shermanii.

scholarly journals Seleção de cepas de Bacillus thuringensis Berliner para o controle de Aedes aegypti Linnaeus

2013 ◽  
Vol 4 (1) ◽  
pp. 78-83
Author(s):  
André Luiz de Almeida Melo ◽  
André Luís Lopes da Silva ◽  
Magda Clara Vieira da Costa Ribeiro ◽  
Mitiyo Fukuda Miyaoka ◽  
Vanete Thomas Soccol ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Aedes Aegypti ◽  
Mosquito Species ◽  
Larval Mortality ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sds Page ◽  
Bacillus Thuringensis ◽  
Two Stages

The present study aimed to select strains of Bacillus thuringiensis with insecticidal activity against Aedes aegypti. It was tested sixteen strains of bacteria, isolated from Paraná state, Brazil, that were used in laboratory assays with mosquito larvae. Tests were carried out in two stages, first one to select the most efficient strains (screening) and second to estimate LC50. The protein profile of the highest toxicity of strain was obtained by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis). The best performance of larval mortality was produced by BR-01 strain, which 96.7% mortality rate, significantly higher than others. In the second part, there was obtained a LC50 of 9.07 µL.L-1 fermented extract. The protein profile revealed many peptides between 15 and 140kDa, similar to that reported in Bacillus thuringiensis ser. israelensis strains which high toxicity to mosquito species.

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