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

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

Bulletin of Entomological Research ◽

10.1017/s0007485314000431 ◽

2014 ◽

Vol 104 (5) ◽

pp. 639-651 ◽

Cited By ~ 3

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.

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

Canadian Journal of Microbiology ◽

10.1139/w08-103 ◽

2009 ◽

Vol 55 (2) ◽

pp. 117-125 ◽

Cited By ~ 9

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.

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New Strategy for Isolating Novel Nematicidal Crystal Protein Genes from Bacillus thuringiensis Strain YBT-1518

Applied and Environmental Microbiology ◽

10.1128/aem.01346-08 ◽

2008 ◽

Vol 74 (22) ◽

pp. 6997-7001 ◽

Cited By ~ 60

Author(s):

Suxia Guo ◽

Mei Liu ◽

Donghai Peng ◽

Sisi Ji ◽

Pengxia Wang ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Meloidogyne Hapla ◽

Microscopy Observation ◽

Root Knot Nematode ◽

Dna Library ◽

Sds Page ◽

New Strategy ◽

Key Steps

ABSTRACT We have developed a strategy for isolating cry genes from Bacillus thuringiensis. The key steps are the construction of a DNA library in an acrystalliferous B. thuringiensis host strain and screening for the formation of crystal through optical microscopy observation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses. By this method, three cry genes—cry55Aa1, cry6Aa2, and cry5Ba2—were cloned from rice-shaped crystals, producing B. thuringiensis YBT-1518, which consists of 54- and 45-kDa crystal proteins. cry55Aa1 encoded a 45-kDa protein, cry6Aa2 encoded a 54-kDa protein, and cry5Ba2 remained cryptic in strain YBT-1518, as shown by SDS-PAGE or microscopic observation. Proteins encoded by these three genes are all toxic to the root knot nematode Meloidogyne hapla. The two genes cry55Aa1 and cry6Aa2 were found to be located on a plasmid with a rather small size of 17.7 kb, designated pBMB0228.

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

International Journal of Applied Sciences and Biotechnology ◽

10.3126/ijasbt.v3i2.12734 ◽

2015 ◽

Vol 3 (2) ◽

pp. 322-329 ◽

Cited By ~ 1

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

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Diversity of Bacillus thuringiensis Strains from Latin America with Insecticidal Activity against Different Mosquito Species

Applied and Environmental Microbiology ◽

10.1128/aem.69.9.5269-5274.2003 ◽

2003 ◽

Vol 69 (9) ◽

pp. 5269-5274 ◽

Cited By ~ 95

Author(s):

Jorge E. Ibarra ◽

M. Cristina del Rincón ◽

Sergio Ordúz ◽

David Noriega ◽

Graciela Benintende ◽

...

Keyword(s):

Latin America ◽

Bacillus Thuringiensis ◽

Insecticidal Activity ◽

Mosquito Species ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Pcr Analysis ◽

Pcr Screening ◽

Management Interventions ◽

Plasmid Profiles

ABSTRACT The characterization of selected Bacillus thuringiensis strains isolated from different Latin America countries is presented. Characterization was based on their insecticidal activity against Aedes aegypti, Culex quinquefasciatus, and Anopheles albimanus larvae, scanning electron microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and plasmid profiles as well as PCR analysis using novel general and specific primers for cry and cyt genes encoding proteins active against mosquitoes (cyt1, cyt2, cry2, cry4A, cry4B, cry10, cry11, cry17, cry19, cry24, cry25, cry27, cry29, cry30, cry32, cry39, and cry40). Strains LBIT315, LBIT348, and IB604 showed threefold higher mosquitocidal activity against A. aegypti and C. quinquefasciatus larvae than B. thuringiensis subsp. israelensis and displayed high similarities with the B. thuringiensis subsp. israelensis used in this study with regard to protein and plasmid profiles and the presence of cry genes. Strain 147-8906 has activity against A. aegypti similar to that of B. thuringiensis subsp. israelensis but has different protein and plasmid profiles. This strain, harboring cry11, cry30, cyt1, and cyt2 genes, could be relevant for future resistance management interventions. Finally, the PCR screening strategy presented here led us to identify a putative novel cry11B gene.

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Crystalline protein profiling and cry gene detection in Bacillus thuringiensis strains isolated during epizootics in Cydia pomonella L.

Biological Letters ◽

10.1515/biolet-2015-0008 ◽

2014 ◽

Vol 51 (2) ◽

pp. 83-92 ◽

Cited By ~ 2

Author(s):

Edyta Konecka ◽

Jakub Baranek ◽

Adam Kaznowski

Keyword(s):

Bacillus Thuringiensis ◽

Polyacrylamide Gel Electrophoresis ◽

Cydia Pomonella ◽

Sodium Dodecyl ◽

Natural Populations ◽

Protein Profiling ◽

Gene Detection ◽

Sds Page ◽

Molecular Masses ◽

Reference Strains

AbstractThe composition of Bacillus thuringiensis crystalline inclusions was characterized in 18 strains: 12 isolates were obtained from the intestinal tract of Cydia pomonella larvae during epizootics, 2 isolates were cultured from Leucoma salicis larvae taken from their natural populations, and 4 reference strains. The number and molecular mass of B. thuringiensis crystalline proteins (Cry and Cyt) was estimated by the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The crystals contained 1-8 proteins with molecular masses of 36-155 kDa. The toxin profiles differed both quantatively and qualitatively. The B. thuringiensis MPU B9 isolate had the highest number and diversity of Cry toxins. The analysis of crystal composition by SDS-PAGE was insufficient to detect groups and subgroups of Cry proteins. We identified 20 groups and 3 subgroups of Cry and Cyt crystalline toxins. Only one epizootic strain harboured cry25. In single reference strains, the cry1H, cry10 and cry25 genes were found. We did not find any correlation between the occurrence of cry genes and electrophoretic protein profiles of crystalline toxins.

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COMPARISON OF POLYPEPTIDE PROFILE OF Trypanosoma evansi ISOLATES FROM INDONESIA AND THEIR RELATION TO BIOTYPE AND SENSITIVITY TO TRYPANOCIDAL

Jurnal Kedokteran Hewan - Indonesian Journal of Veterinary Sciences ◽

10.21157/j.ked.hewan.v12i2.11486 ◽

2018 ◽

Vol 12 (2) ◽

Author(s):

Ichwan Yuniarto ◽

Didik T Subekti ◽

Umi Cahyaningsih ◽

Fadjar Satrija

Keyword(s):

Biological Diversity ◽

Protein Profile ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Trypanosoma Evansi ◽

Central Kalimantan ◽

Central Java ◽

Sds Page ◽

The Difference ◽

Polypeptide Profile

This study aimed to determine whether the variant or biotype of Trypanosoma evansi can be seen from their polypeptide profiles using 12%sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) stained with Brilliant Blue Commasie. The results generally showed thatthe molecular weight (MW) of polypeptides from nine isolates from East Java, Central Java, Banten, South Kalimantan, Central Kalimantan, andLampung provinces were in the range of 85.46 to 15.76 kD and each isolate has different polypeptide profile. Isolates A13 and A14 were isolatedfrom the same place but have different polypeptide profiles. Likewise, isolates S13 and S18 also have different polypeptide profiles despite beingisolated from the same place at the same time. On the other hand, isolate 372, 87, and 06 have different protein profiles but was classified in thesame biotype namely biotype I. Generally, the difference in protein profile actually more related to the biological diversity of the metabolism ofeach Trypanosoma evansi isolate from Indonesia.

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Characterization and selection of Bacillus thuringiensis isolates effective against Sitophilus oryzae

Scientia Agricola ◽

10.1590/s0103-90162010000400015 ◽

2010 ◽

Vol 67 (4) ◽

pp. 472-478 ◽

Cited By ~ 3

Author(s):

Najara da Silva ◽

Ana Maria Guidelli Thuler ◽

Irlan Leite de Abreu ◽

Camila Chiaradia Davolos ◽

Ricardo Antonio Polanczyk ◽

...

Keyword(s):

Bacillus Thuringiensis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Sitophilus Oryzae ◽

Genetic Material ◽

Control Agent ◽

Integrate Pest Management ◽

Sds Page ◽

Pathogenicity Tests ◽

Bacterium Bacillus

The entomopathogenic bacterium Bacillus thuringiensis is a control agent with toxic and environmental characteristics that allows the control of pest insects according to the Integrate Pest Management (IPM) precepts. In order to find new strains, potentially toxic to Sitophilus oryzae L. 1763 (Coleoptera: Curculinidae), 1.073 strains of B. thuringiensis from parts of Brazil were used. Genetic material was extracted with InstaGene Matrix kit, used for the amplification of sequences in Polymerase chain reaction (PCR), and viewed in 1.5% agarose gel. The gene cry35Ba class was represented by 60 B. thuringiensis isolates (5.6%), which were then subjected to bioassays with S. oryzae larvae. Among the isolates studied, four caused more than 50% mortality in pathogenicity tests, and the isolates 544 and 622 were the most virulent, as determined by CL50 estimates. The four toxic isolates had spherical, bi-pyramidal and cuboid crystals, and a 44-kDa protein was found in sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE), which coded for the product of cry35Ba genes. These data demonstrate the potential of B. thuringiensis for the management of S. oryzae larvae.

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Perbandingan Metode Hierarchical Cluster Analysis untuk Analisis Keragaman Hayati Trypanosoma evansi dari Indonesia Berdasarkan Profil Protein (COMPARISON OF HIERARCHICAL CLUSTER ANALYSIS METHODS FOR BIODIVERSITY ANALYSIS OF TRYPANOSOMA EVANSI

jurnal veteriner ◽

10.19087/jveteriner.2017.18.4.516 ◽

2018 ◽

Vol 18 (4) ◽

pp. 516

Author(s):

Didik Tulus Subekti ◽

Ichwan Yuniarto ◽

Sulinawati Sulinawati

Keyword(s):

Cluster Analysis ◽

Hierarchical Cluster Analysis ◽

Binary Data ◽

Protein Profile ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Hierarchical Cluster ◽

Trypanosoma Evansi ◽

Protein Profiles ◽

Sds Page

Hierarchical Clustering Analysis (HCA) has long been known to be useful for the analysis of biodiversity of microorganisms based on SDSPAGE protein profile (sodium dodecyl sulfate polyacrylamide gel electrophoresis). However, varying methods of HCA consequently produce variability of analysis results and interpretations. Therefore, it is necessary to evaluate and further determine the most appropriate method which could described the biodiversity based on protein profiles of T.evansi isolates from Indonesia. Eleven isolates of T.evansi from different geographic locations were run on SDS PAGE. Furthermore, SDS PAGE protein profiles from eleven isolates were converted into binary data and analyzed using five different methods of HCA i.e. Average Linkage, Complete Linkage, Single Linkage, Ward Linkage and McQuitty Linkage, respectively.Data were also analyzed by multidimensional scaling (MDS) and densitogram. The analysis showed that the dendrogram constructed with Ward Linkage gives the best results and corresponding with densitogram, MDS and able to describe the geographical origins of isolates.

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Analysis of the protein profile of the venoms of snakes Bothrops asper, Bothrocophias myersi and Crotalus durissus from the Colombian Andean Region obtained by RP-HPLC

Revista Colombiana de Biotecnología ◽

10.15446/rev.colomb.biote.v23n1.94211 ◽

2021 ◽

Vol 23 (1) ◽

pp. 24-31

Author(s):

Stefano Scovino Loboguerrero ◽

Karen Sarmiento ◽

Carlos Galvis ◽

Ana Lucía Castiblanco ◽

Fabio Aristizabal

Keyword(s):

Protein Profile ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Complex Mixture ◽

Clinical Syndrome ◽

Andean Region ◽

Rp Hplc ◽

Sds Page ◽

Bothrops Asper ◽

Crotalus Durissus

Snake venoms comprise a highly complex mixture of proteins, and there is also a high interspecific and intraspecific variability in their composition, even in the same region. Our aim was to compare the composition of the venoms of Bothrocophias myersi, Crotalus durissus and Bothrops asper, snakes from the Andean region in Colombia by Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC). The venoms were given to the research group under an agreement with the Fundación Zoológica de Cali. The venoms pool was obtained by manual extraction, lyophilized and refrigerated. The protein found in the venoms was quantified by spectrophotometry using the Bradford and Lowry methods and direct measurement by Nanodrop®. The protein composition was stablished by RP-HPLC, using a Lichosper 100 RP, C18 column (250X4 mm) with a pore size of 5µm, as well as by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). The highest quantity of protein was found in the venom of B. myersi (108,6 mg/mL) followed by C. durissus (78,1 mg/mL) and B. asper (74.1 mg/mL). All venoms showed bands of 15 and 50 KDa by SDS-PAGE; The most important finding is the abundance of PLA2 and svMP in the venom of B. myersi. Chromatographic analyses revealed a very similar venom composition profile, but also certain differences in toxins abundance. We conclude that the process of separating the venom proteins by RP-HPLC and SDS-PAGE are very important as a first step to know the venoms profiles, which in turn could allow medical staff to elucidate the clinical syndrome produced by snakebites.

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