Hydrogen peroxide agarose gels for electrophoretic analysis of RNA

2017 ◽  
Vol 534 ◽  
pp. 24-27 ◽  
Author(s):  
Renu Pandey ◽  
Daman Saluja
Keyword(s):  
Hydrogen Peroxide ◽  
Electrophoretic Analysis ◽  
Agarose Gels

Comparison of enzymatic antioxidant defence systems in different metabolic types of yeasts

2008 ◽  
Vol 54 (11) ◽  
pp. 957-963 ◽  
Author(s):  
Dafinka I. Koleva ◽  
Ventsislava Y. Petrova ◽  
Anna V. Kujumdzieva
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Hydrogen Peroxide ◽  
Taxonomic Status ◽  
Rhodotorula Glutinis ◽  
Electrophoretic Analysis ◽  
Mobility Pattern ◽  
Antioxidant Defence ◽  
Sod Activity ◽  
Fermentative Capacity

The enzymatic defence system in the 2 yeasts Kluyveromyces marxianus and Rhodotorula glutinis , differing in their mode of oxygen uptake and energy generation, was characterized and compared with the well-studied facultatively fermentative Crabtree-positive Saccharomyces cerevisiae strain. Twofold higher superoxide dismutase (SOD) and catalase activities were detected in K. marxianus and R. glutinis when cells were cultured on glucose. Further increases of 10%–15% in SOD activity and 30%–50% in catalase were measured in all studied yeasts strains after transfer to media containing ethanol. An evaluation of the ratio of Cu/Zn SOD / Mn SOD was performed as a measure of the oxidative metabolism. A 20% decrease was observed when the respiratory source of energy was ethanol, with the lowest ratio being observed for the oxidative type of K. marxianus yeasts. Electrophoretic analysis revealed that all tested strains possess active Cu/Zn and Mn SODs. A reverse electrophoretic mobility pattern of K. marxianus and R. glutinis SOD enzymes was observed in comparison with the same couple in S. cerevisiae. The investigation of electrophoretic profile of catalase enzymes showed that alongside their different taxonomic status and fermentative capacity, all tested strains possess 2 separate catalases. The role of antioxidant enzymes in preventing oxidant-induced cytotoxicity (treatment with hydrogen peroxide, paraquat, and menadione) was shown.

scholarly journals Electrophoretic analysis of Histoplasma capsulatum chromosomal DNA.

1989 ◽  
Vol 9 (3) ◽  
pp. 983-987 ◽  
Author(s):  
P E Steele ◽  
G F Carle ◽  
G S Kobayashi ◽  
G Medoff
Keyword(s):  
Gel Electrophoresis ◽  
Histoplasma Capsulatum ◽  
Electrophoretic Analysis ◽  
Considerable Variability ◽  
Dna Molecules ◽  
Homogeneous Electric Field ◽  
Chromosomal Dna ◽  
Experimental Conditions ◽  
Agarose Gels ◽  
Minimum Estimate

Seven chromosome-sized DNA molecules in the Downs strain of Histoplasma capsulatum were resolved by using chromosome-specific DNA probes in blot hybridizations of contour-clamped homogeneous electric field (CHEF) and field-inversion gel electrophoresis (FIGE) agarose gels. The sizes of the chromosomal DNA bands extended from that of the largest Saccharomyces cerevisiae chromosome to beyond that of the Schizosaccharomyces pombe chromosomes. Under our experimental conditions, the order of the five largest DNA bands was inverted in the FIGE gel relative to the CHEF gel, demonstrating a characteristic of FIGE whereby large DNA molecules may have greater rather than lesser mobility with increasing size. Comparison of the Downs strain with other H. capsulatum strains by CHEF and FIGE analysis revealed considerable variability in band mobility. The resolution of seven chromosome-sized DNA molecules in the Downs strain provides a minimum estimate of the chromosome number.

Jelly plug dissolution in Discoglossus pictus eggs (Anura) involves peroxidase-like activity and oxidative opening of disulphide bonds

Zygote ◽  
1993 ◽  
Vol 1 (1) ◽  
pp. 61-69 ◽  
Author(s):  
G. Pitari ◽  
S. Dupré ◽  
C. Fusco ◽  
G. Maurizi ◽  
C. Campanella
Keyword(s):  
Hydrogen Peroxide ◽  
Peroxidase Activity ◽  
Electrophoretic Analysis ◽  
Ringer Solution ◽  
Cysteic Acid ◽  
Jelly Coat ◽  
Discoglossus Pictus ◽  
Disulphide Bonds ◽  
Sds Page

SummaryIn amphibian eggs the formation of a capsular chamber is one of the most striking events occurring either upon oviposition or after fertilisation. In the egg of the anuran Discoglossus pictus a capsular chamber forms following fertilisation or activation; the egg with its vitelline envelope rotates in this chamber according to gravity. Previous work showed that the chamber is the product of plug dissolution. The plug is a lens-shaped jelly coat, typical of Discoglossus, covering only part of the animal hemisphere. Its dissolution is caused by material released from the egg about 15 min after fertilisation through exocytosis of at least two types of vacuoles. Liquefaction of the plug correlates with the reduction of disulphide bonds present in the jelly matrix. In this study we investigated the nature of the substances released from the egg and some changes occurring in the plug during liquefaction. SDS-PAGE showed that the proteic profile of the plug changes dramatically after fertilisation, confirming proteic cleavage in the plug matrix during its dissolution. Through in vitro tests and electrophoretic analysis of the Ringer solution in which the egg exudate was collected, an increase in the activity of the solution was determined in the presence of hydrogen peroxide, and peroxidase activity was depicted in the egg exudate. The presence of free thiol groups and cysteic acid residues (or cysteine sulphinic acid) in the plugs of activated eggs was established, suggesting that during plug dissolution some disulphide bonds are oxidatively opened. This suggests that enzyme(s) with peroxidase activity are released following fertilisation. We surmise that such enzymes are contained in the intraovular vacuoles the exocytosis of which triggers the onset of plug liquefaction. The possible release of hydrogen peroxide from the egg is discussed.

Plasmid recombination intermediates generated in a Saccharomyces cerevisiae cell-free recombination system

1985 ◽  
Vol 5 (9) ◽  
pp. 2361-2368
Author(s):  
L S Symington ◽  
P Morrison ◽  
R Kolodner
Keyword(s):  
Electron Microscopy ◽  
Saccharomyces Cerevisiae ◽  
Electrophoretic Mobility ◽  
Electrophoretic Analysis ◽  
Individual Band ◽  
Agarose Gels ◽  
Cell Extracts ◽  
Recombination System ◽  
Plasmid Recombination

We have developed an assay utilizing Saccharomyces cerevisiae cell extracts to catalyze recombination in vitro between homologous plasmids containing different mutant alleles of the tet gene. Electrophoretic analysis of product DNA indicated that a number of novel DNA species were formed during the reaction. These species migrated through agarose gels as distinct bands with decreased electrophoretic mobility compared with the substrate DNA. The DNA from each individual band was purified and shown to be enriched 5- to 100-fold for tetracycline-resistant recombinants by using a transformation assay. The structure of the DNA molecules present in these bands was determined by electron microscopy. Recombination between circular substrates appeared to involve the formation and processing of figure-eight molecules, while recombination between circular and linear substrates involved the formation of molecules in which a circular monomer had a monomer-length linear tail attached at a region of homology.

scholarly journals Plasmid recombination intermediates generated in a Saccharomyces cerevisiae cell-free recombination system.

1985 ◽  
Vol 5 (9) ◽  
pp. 2361-2368 ◽  
Author(s):  
L S Symington ◽  
P Morrison ◽  
R Kolodner
Keyword(s):  
Electron Microscopy ◽  
Saccharomyces Cerevisiae ◽  
Electrophoretic Mobility ◽  
Electrophoretic Analysis ◽  
Individual Band ◽  
Agarose Gels ◽  
Cell Extracts ◽  
Recombination System ◽  
Plasmid Recombination

We have developed an assay utilizing Saccharomyces cerevisiae cell extracts to catalyze recombination in vitro between homologous plasmids containing different mutant alleles of the tet gene. Electrophoretic analysis of product DNA indicated that a number of novel DNA species were formed during the reaction. These species migrated through agarose gels as distinct bands with decreased electrophoretic mobility compared with the substrate DNA. The DNA from each individual band was purified and shown to be enriched 5- to 100-fold for tetracycline-resistant recombinants by using a transformation assay. The structure of the DNA molecules present in these bands was determined by electron microscopy. Recombination between circular substrates appeared to involve the formation and processing of figure-eight molecules, while recombination between circular and linear substrates involved the formation of molecules in which a circular monomer had a monomer-length linear tail attached at a region of homology.

Electrophoretic analysis of Histoplasma capsulatum chromosomal DNA

1989 ◽  
Vol 9 (3) ◽  
pp. 983-987
Author(s):  
P E Steele ◽  
G F Carle ◽  
G S Kobayashi ◽  
G Medoff
Keyword(s):  
Gel Electrophoresis ◽  
Histoplasma Capsulatum ◽  
Electrophoretic Analysis ◽  
Considerable Variability ◽  
Dna Molecules ◽  
Homogeneous Electric Field ◽  
Chromosomal Dna ◽  
Experimental Conditions ◽  
Agarose Gels ◽  
Minimum Estimate

Seven chromosome-sized DNA molecules in the Downs strain of Histoplasma capsulatum were resolved by using chromosome-specific DNA probes in blot hybridizations of contour-clamped homogeneous electric field (CHEF) and field-inversion gel electrophoresis (FIGE) agarose gels. The sizes of the chromosomal DNA bands extended from that of the largest Saccharomyces cerevisiae chromosome to beyond that of the Schizosaccharomyces pombe chromosomes. Under our experimental conditions, the order of the five largest DNA bands was inverted in the FIGE gel relative to the CHEF gel, demonstrating a characteristic of FIGE whereby large DNA molecules may have greater rather than lesser mobility with increasing size. Comparison of the Downs strain with other H. capsulatum strains by CHEF and FIGE analysis revealed considerable variability in band mobility. The resolution of seven chromosome-sized DNA molecules in the Downs strain provides a minimum estimate of the chromosome number.

scholarly journals Actin and Myosin from Platelets and Muscle are Substrates for Fibrinoligase

1977 ◽  
Author(s):  
Isaac Cohen ◽  
Joyce Bruner-Lorand ◽  
Tikoes A. Blankenberg ◽  
Laszlo Lorand
Keyword(s):  
Skeletal Muscle ◽  
Factor Xiii ◽  
Sodium Dodecyl ◽  
Electrophoretic Analysis ◽  
Human Platelets ◽  
Light Chains ◽  
Agarose Gels ◽  
Heavy Chains ◽  
Cross Links ◽  
Muscle Myosin

Fibrinoligase (thrombin and calcium activated factor XIII) from human plasma catalyzes the incorporation of dansylcadaverine and 14C-putrescine into myosin and actin prepared from either human platelets or rabbit skeletal muscle. An average of 3 to 4 moles of amine are incorporated per mole of myosin of either type when the enzyme is used under saturating conditions. The rate of incorporation is 1.4 to 2 fold faster for filamentous than for soluble myosin.Electrophoretic analysis on Polyacrylamide and agarose gels in the presence of sodium dodecyl sulfate and 2-mercaptoethanol was used for locating the acceptor sites and for the detection of possible cross-links. Both heavy and light chains of platelet as well as muscle myosin incorporate dansylcadaverine and 14C-putrescine. However, in quantitative terms, the incorporation into the light chains of either type is only a small fraction of that incorporated into the heavy chains. Fibrinoligase also catalyzes the formation of highly cross-linked polymers in both myosin and actin (platelet or muscle) indicating the presence of acceptor and donor sites in the two contractile proteins.

Peroxidase Localization in Hartmanella Trophozoites

1971 ◽  
Vol 29 ◽  
pp. 346-347 ◽  
Author(s):  
George E. Childs ◽  
Joseph H. Miller
Keyword(s):  
Hydrogen Peroxide ◽  
Ultrastructural Localization ◽  
Pathogenic Strain ◽  
Oxidative Enzymes ◽  
Differential Centrifugation ◽  
Electron Microscopic ◽  
Microscopic Studies ◽  
The Subject ◽  
Axenic Cultures

Biochemical and differential centrifugation studies have demonstrated that the oxidative enzymes of Acanthamoeba sp. are localized in mitochondria and peroxisomes (microbodies). Although hartmanellid amoebae have been the subject of several electron microscopic studies, peroxisomes have not been described from these organisms or other protozoa. Cytochemical tests employing diaminobenzidine-tetra HCl (DAB) and hydrogen peroxide were used for the ultrastructural localization of peroxidases of trophozoites of Hartmanella sp. (A-l, Culbertson), a pathogenic strain grown in axenic cultures of trypticase soy broth.

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