Immunoblot patterns of Giardia duodenalis isolates from different hosts and geographical locations

1990 ◽  
Vol 36 (1) ◽  
pp. 42-46 ◽  
Author(s):  
M. Forrest ◽  
J. Isaac-Renton ◽  
W. Bowie
Keyword(s):  
Key Words ◽  
Banding Pattern ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Giardia Duodenalis ◽  
Immunoblot Analysis ◽  
Polyacrylamide Gels ◽  
Banding Patterns ◽  
Antigenic Reactivity ◽  
Geographical Locations

Eighteen isolates of Giardia duodenalis from animal and human sources were studied for protein differences by polyacrylamide gel electrophoresis and for antigenic differences by immunoblot analysis. The polyacrylamide gels showed that whilst the isolates were for the most part homogeneous in their protein banding patterns, some isolates did show some differences. The immunoblot analysis yielded many bands, including prominent bands of 32 and 66 kilodaltons. Five of the six isolates that showed differences in protein banding pattern also showed differences in antigenic reactivity. Our findings suggest that differences can be seen with the use of immunoblotting and that this technique is a tool that may be useful for isolate differentiation when used in conjunction with other techniques. Key words: Giardia, giardiasis, characterization, immunoblot.

Isoenzyme analysis of teliospores from species of Anthracoidea parasitic on Carex species

1993 ◽  
Vol 71 (11) ◽  
pp. 1406-1413 ◽  
Author(s):  
Vanamo Salo ◽  
Robin Sen
Keyword(s):  
Genetic Variation ◽  
Banding Pattern ◽  
Gel Electrophoresis ◽  
Cell Walls ◽  
Morphological Character ◽  
Polyacrylamide Gel Electrophoresis ◽  
Isoenzyme Analysis ◽  
Banding Patterns ◽  
Multiple Loci ◽  
Smut Spores

The smut sori of five Anthracoidea species, A. echinospora, A. heterospora, A. buxbaumii, A. limosa, and A. paniceae, removed from infected inflorescences of 5 Carex host plant species collected from 17 natural Finnish populations, were surveyed by isozyme analysis. The protein extracts of individual sori, containing teliospores (smut spores), together with uninfected utricles, were subjected to polyacrylamide gel electrophoresis and stained for esterase and four peptidase enzymes. Single sori, representing individual infection units, were sufficient to reveal considerable esterase and peptidase isozyme activities, whereas little or no activity was seen in the control utricle extracts. High levels of genetic variation as expressed by the detection of multiple loci and allelism were recorded between species and various populations, but variation within populations was lower. Peptidase banding patterns, in particular, proved to be useful in differentiating species: A. heterospora and A. echinospora, parasitic on the same plant, were clearly distinguished by their isozyme patterns, and the three remaining smuts belonging to the subgenus Proceres could be separated from one another. One morphological character, presence of numerous thick internal swellings in the cell walls of teliospores, seemed to correlate well with banding pattern differences in A. heterospora. Key words: isozymes, electrophoresis, Anthracoidea, Carex, chemotaxonomy.

Electrophoretic evidence supporting self-incompatibility in Lotus corniculatus

1980 ◽  
Vol 58 (6) ◽  
pp. 712-716 ◽  
Author(s):  
Shirley Dobrofsky ◽  
W. F. Grant
Keyword(s):  
Gel Electrophoresis ◽  
Seed Set ◽  
Polyacrylamide Gel Electrophoresis ◽  
Lotus Corniculatus ◽  
Polyacrylamide Gel ◽  
Self Incompatibility ◽  
Banding Patterns ◽  
Protein Subunits ◽  
Self Pollination

Self-incompatibility, a prefertilization event, and self-sterility, a postfertilization event, have both been suggested as causes for differences in seed set between cross- and self-pollinated florets in Lotus corniculatus L. Ovary protein subunits of selfed, crossed, and unpollinated florets of L. corniculatus cv. Mirabel were studied using polyacrylamide gel electrophoresis. Banding patterns differed for all three conditions. Ovary protein differences were found prior to the time fertilization is known to occur, thereby providing evidence that self-incompatibility is at least partially responsible for the reduced seed set after self-pollination.

scholarly journals Involement of an acrosinlike proteinase in the sulfhydryl-induced degradation of rabbit sperm nuclear protamine

1980 ◽  
Vol 85 (1) ◽  
pp. 116-121 ◽  
Author(s):  
BR Zirkin ◽  
TSK Chang ◽  
J Heaps
Keyword(s):  
Proteolytic Activity ◽  
Banding Pattern ◽  
Basic Protein ◽  
Polyacrylamide Gels ◽  
Banding Patterns ◽  
Pattern Characteristic ◽  
Sperm Nuclei ◽  
Triton X ◽  
Esterase Inhibitor

Previous studies demonstrated that proteolytic activity is associated with isolated rabbit sperm nuclei and is responsible for the degradation of nuclear protamine that occurs during thiol-induced in vitro decondensation of the nuclei (Zirkin and Chang, 1977; Chang and Zirkin, 1978). In this study, we present the results of experiments designed to characterize this proteolytic activity. Basic protein isolated from rabbit sperm nuclei incubated with 5 mM dithiothreitol (DTT) and 1 percent Triton X-100 for increasing periods of time exhibited progressively faster migrating bands on acid-urea polyacrylamide gels, reflection the progressive degradation of protamine. Ultimately, a specific and characteristic peptide banding pattern resulted. When sperm nuclei were treated with the esterase inhibitor nitrophenyl-p-guanidino benzoate (NPGB) to inhibit the nuclear-associated proteolytic activity and then incubated with one of several exogenous proteinases in addition to DTT and Triton X-100, characteristic peptide banding patterns were seen for each exogenous proteinase employed. For trypsin, chymotrypsin, pronase, and papain, the peptide banding patterns differed from one another and from the pattern characteristic of protamine degradation by the nuclear-associated proteinase. By contrast, when rabbit acrosin served as the exogenous proteinase, the peptide banding pattern seen was identical to the pattern characteristic of the nuclear-associated proteinase. These results demonstrate directly that the proteinase associated with rabbit sperm nuclei and involved in sperm nuclear decondensation in vitro is acrosinlike.

The Electrophoretic Separation of Curved Cisplatin-Modifled DNA Fragments on Polyacrylamide Gels Is Dependent on the Voltage Gradient

1998 ◽  
Vol 53 (9-10) ◽  
pp. 921-923
Author(s):  
Julia Yaneva ◽  
Jordanka Zlatanova
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Threshold Level ◽  
Voltage Gradient ◽  
Dna Fragments ◽  
Electrophoretic Separation ◽  
Polyacrylamide Gels ◽  
Curved Dna ◽  
Electrophoretic Behavior ◽  
Param Eters

Polyacrylamide gel electrophoresis has been widely used to study DNA fragments containing sequence-dependent curvature. The anomalous electrophoretic behavior of curved DNA fragments on such gels allows their separation from straight fragments of the same length. Here we demonstrate that polyacrylamide gels can be successfully used to resolve DNA fragments modified at a single site by the antitumor drug cis-diamminedichloroplatinum(II) (cis-DDP, cisplatin) from their unmodified counterparts. However, the resolution strongly depends on the voltage gradient, being completely lost when it drops below a certain threshold level. The param eters of the electric field do not affect separation of ‘normal’ DNA fragments of comparable length.

An analysis of the proteinases ofTrichomonas vaginalisby polyacrylamide gel electrophoresis

Parasitology ◽  
1983 ◽  
Vol 86 (1) ◽  
pp. 1-6 ◽  
Author(s):  
G. H. Coombs ◽  
M. J. North
Keyword(s):  
Gel Electrophoresis ◽  
Trichomonas Vaginalis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Cysteine Proteinases ◽  
Polyacrylamide Gels

SUMMARYThe proteinases ofTrichomonas vaginalishave been analysed by electrophoresis in polyacrylamide gels containing denatured haemoglobin. Seven bands of activity were detected indicating multiple proteinases. All of the enzymes were stimulated by 1 mM dithiothreitol and had inhibitor sensitivities characteristic of cysteine proteinases. The enzymes differed significantly, however, with respect to pH optima and relative sensitivities to inhibitors.

Isozymes as markers for identification of tissue culture and greenhouse-grown strawberry cultivars

1991 ◽  
Vol 71 (4) ◽  
pp. 1195-1201 ◽  
Author(s):  
N. S. Nehra ◽  
K. K. Kartha ◽  
C. Stushnoff
Keyword(s):  
Tissue Culture ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Meristem Culture ◽  
Banding Patterns ◽  
Extraction Buffer ◽  
Leaf Tissues ◽  
The Stability ◽  
Strawberry Cultivars

Polyacrylamide gel electrophoresis (PAGE) was used for analysis of isozyme banding patterns of leucine aminopeptidase (LAP), phosphoglucomutase (PGM), phosphoglucoisomerase (PGI), esterase (EST) and 6-phosphogluconate dehydrogenase (6-PGD) in strawberry leaves. The extracts prepared from young leaf tissues using polytron homogenization and an extraction buffer containing 15 mg ml−1 dithiothreitol (DTT) and 10% insoluble polyvinylpyrrolidone (PVP-6755) gave best resolution for these enzymes. The influence of plant age and various growing environments on the stability of isozyme phenotypes was examined. The isozyme banding patterns of 6-PGD were found to vary with the change in growing environment as well as age of the plants. EST produced different banding patterns in greenhouse and tissue culture leaves. However, the isozyme phenotypes of LAP, PGM and PGI remained stable under all the conditions tested. Using a combination of these three stable enzymes, it was possible to distinguish eight strawberry cultivars under both tissue culture and greenhouse conditions. Key words: Fragaria × ananassa Duch., meristem culture, polyacrylamide gel electrophoresis

Protein composition of Methanococcus thermolithotrophicus flagella

1992 ◽  
Vol 38 (11) ◽  
pp. 1162-1166 ◽  
Author(s):  
Alla S. Kostyukova ◽  
Georgi M. Gongadze ◽  
Anna Ya. Obraztsova ◽  
Konstantin S. Laurinavichus ◽  
Oleg V. Fedorov
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Key Words ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Composition ◽  
Complete Removal ◽  
Methanococcus Thermolithotrophicus ◽  
Molecular Weights ◽  
Dodecyl Sulfate

Sodium dodecyl sulfate – polyacrylamide gel electrophoresis of flagella from the thermophilic methanogen Methanococcus thermolithotrophicus indicated that they were composed of three major proteins, with molecular weights of 62 000,44 000, and 26 000, whereas all previously studied flagella of mesophilic methanogens consisted of two subunits. Proteins were isolated by preparative electrophoresis followed by complete removal of sodium dodecyl sulfate and their renaturation. It was shown that at least two of the proteins contain a thermostable domain whose complete denaturation proceeds only upon prolonged boiling in the presence of sodium dodecyl sulfate. Key words: flagellin, thermostability, archaebacteria, Methanococcus thermolithotrophicus.

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