scholarly journals 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

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.

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.

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 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 Characterisation and Biological Activity of Stingray Venom (HimanturaImbricata)

2021 ◽  
Vol 7 (03) ◽  
pp. 35-43
Author(s):  
Selvaraj Uthra, Ramachandiran Sivaramakrishnan ◽  
Thirukumar Sangeshwari, Ganapathy Sivaranjani ◽  
Shankar Kanchana and Muthuvel Arumugam
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Coagulation Factors ◽  
Freshwater Ecosystem ◽  
Plasma Coagulation ◽  
Rp Hplc ◽  
Sds Page ◽  
Ftir Spectral Analysis ◽  
Characterization Studies ◽  
The Common

Stingrays envenomation in humanswere the common accident in the marine and freshwater ecosystem. To determine such effect species Himantura imbricata have been used to elucidate Hemolytic activity, Plasma Coagulation, Fibrin coagulation, Fibrinolytic activity, Activated Partial Thromboplastin Time (APTT),and Prothrombin Time (PT) effects were studied. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Amino acid analysis by RP-HPLC, FTIR spectral analysis and SEM were carried for characterization studies. The results show the of plasma coagulation, fibrin coagulation, of this stingray venom delays the coagulation of citrated plasma. APTT and PT results showed intrinsic and extrinsic coagulation factors that were responsible in time delay when compared with the control. Moreover, these biological experiments and characterization studies aided in understanding the envenoming factors and these results might base the development of treatments for complex diseases.

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.

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

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.

scholarly journals 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

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.

Evidence that the bovine ovary secretes large amounts of monomeric inhibin α subunit and its isolation from bovine follicular fluid

1989 ◽  
Vol 2 (3) ◽  
pp. 189-200 ◽  
Author(s):  
P. G. Knight ◽  
A. J. Beard ◽  
J. H. M. Wrathall ◽  
R. J. Castillo
Keyword(s):  
Follicular Fluid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Ovarian Vein ◽  
Α Subunit ◽  
Amino Terminal ◽  
Rp Hplc ◽  
Reducing Conditions ◽  
Sds Page ◽  
Bovine Ovary

ABSTRACT Analysis of bovine follicular fluid (FF) using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) coupled with a sensitive immunoblotting procedure resolved several components that were immunoreactive with an antiserum directed against the n-terminus of the α subunit of human inhibin (hIα(1–32)). Under non-reducing conditions, three intensely stained bands having apparent Mr values of 116 000, 44 000 and 25 000 were present, whilst under reducing conditions only two intensely stained bands (Mr 43 000 and 21 000) were detected. The Mr 44 000 and 25 000 immunoreactive forms (non-reducing conditions) were also demonstrated in bovine utero-ovarian vein and peripheral venous plasma after subjecting samples (40 ml) to immunoaffinity concentration using Sepharose beads coupled to anti-hIα(1–32), SDS-PAGE and immunoblotting. The same approach revealed the presence of the smaller (Mr 25 000) form in bovine granulosa cell-conditioned culture medium (GCCM). Gel-permeation chromatography (Sephacryl S-200), immunoaffinity chromatography (Sepharose-anti-hIα(1–32)) and reversed-phase high-performance liquid chromatography (RP-HPLC; C18 and C8 columns) were employed to isolate from bFF (30 ml, 195 g protein) 750 μg protein which appeared essentially homogeneous by RP-HPLC and SDS-PAGE and had an Mr of 25 000 (non-reducing conditions)/21 000 (reducing conditions), identical to that of the immunoreactive component of lowest Mr found in bovine FF, utero-ovarian vein plasma, peripheral plasma and GCCM. The isolated material was highly immunoreactive with antisera against both hIα(1–32) and purified Mr 32 000 bovine inhibin but was devoid of biological activity when tested in a rat pituitary cell inhibin bioassay. Amino-terminal analysis revealed an amino acid sequence (residues 1–14) identical to that reported elsewhere for the α subunit (Mr 20 000/21 000) of bovine inhibin. In conclusion, the present study has revealed that the bovine ovary secretes considerable quantities of monomeric inhibin α subunit. The unexpected presence of this material in peripheral blood is likely to hinder attempts to obtain physiologically relevant data on circulating levels of inhibin in cattle using conventional radioimmunoassays.

scholarly journals Using SDS-PAGE and ISSR as biochemical markers for assessment the genetic similarity and protein analysis of some cyprinid fish species

Genetika ◽  
2020 ◽  
Vol 52 (1) ◽  
pp. 161-175
Author(s):  
Ali Abu-Almaaty ◽  
Iman Bahgat ◽  
Zaineb Al-Tahr
Keyword(s):  
Fish Species ◽  
Dna Analysis ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Analysis ◽  
Cyprinid Fish ◽  
Brachydanio Rerio ◽  
Sds Page ◽  
Issr Primers

Genetics similarity and protein analysis of some cyprinid fish were studied using Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Inter-simple sequence repeated (ISSR Markers). Species Pethia nigrofasciatus, Barbonymus schwanenfeldii, Puntius tetrazone and Brachydanio rerio were collected from the fish farms in Damietta to study the genetic variability among them. Eleven ISSR primers were tested to assess the effectiveness of ISSR analysis in discriminating among the four applied fish species. We observed varied size of amplified products depending upon the sequence of ISSR primers and genotypes used. A total of 131 discrete amplified products were obtained (size 79 to 1185 bp approximately) with polymorphism 95%. Out of 131 products, 63 bands were species specific markers indicating high level polymorphism among species. The highest and lowest number of ISSR bands detected for primers ISSR 15 and ISSR16 was 18 and 17 respectively. 7 bands were most relevant as found monomorphic in all four species of family: Cyprinidae. Highest similarity observed between Pethia nigrofasciatus and Barbonymus schwanenfeldii 80% and lowest similarity was between Pethia nigrofasciatus and Brachydanio rerio 31 %. The protein analysis by SDS-PAGE produced 29 bands of molecular weight ranging from 11 to 132 KD with polymorphism 14%. This study concludes that ISSR-based DNA analysis bands and protein profile in muscles from Pethia nigrofasciatus and Barbonymus schwanenfeldii are the most closest species compared molecularly compared with other species used in this study.

Heterozygous Abnormal Fibrinogen Osaka III with the Replacement of γ Arginine-275 by Histidine Has an Apparently Higher Molecular Weight γ-Chain Variant

1992 ◽  
Vol 68 (05) ◽  
pp. 534-538 ◽  
Author(s):  
Nobuhiko Yoshida ◽  
Shingi Imaoka ◽  
Hajime Hirata ◽  
Michio Matsuda ◽  
Shinji Asakura
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Tetraacetic Acid ◽  
Fibrin Monomer ◽  
Sds Page ◽  
Thrombotic Tendency ◽  
Chain Variant

SummaryCongenitally abnormal fibrinogen Osaka III with the replacement of γ Arg-275 by His was found in a 38-year-old female with no bleeding or thrombotic tendency. Release of fibrinopeptide(s) by thrombin or reptilase was normal, but her thrombin or reptilase time in the absence of calcium was markedly prolonged and the polymerization of preformed fibrin monomer which was prepared by the treatment of fibrinogen with thrombin or reptilase was also markedly defective. Propositus' fibrinogen had normal crosslinking abilities of α- and γ-chains. Analysis of fibrinogen chains on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system of Laemmli only revealed the presence of abnormal γ-chain with an apparently higher molecular weight, the presence of which was more clearly detected with SDS-PAGE of fibrin monomer obtained by thrombin treatment. Purified fragment D1 of fibrinogen Osaka III also seemed to contain an apparently higher molecular weight fragment D1 γ remnant on Laemmli gels, which was digested faster than the normal control by plasmin in the presence of [ethy-lenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA).

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