Using Two-Dimensional Intact Mitochondrial DNA (mtDNA) Agarose Gel Electrophoresis (2D-IMAGE) to Detect Changes in Topology Associated with Mitochondrial Replication, Transcription, and Damage

2020 ◽  
pp. 25-42
Author(s):  
Jill E. Kolesar ◽  
Brett A. Kaufman
Keyword(s):  
Mitochondrial Dna ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Mitochondrial Replication

Synthesis and processing of kinetoplast DNA minicircles in Trypanosoma equiperdum

1989 ◽  
Vol 9 (8) ◽  
pp. 3212-3217
Author(s):  
K A Ryan ◽  
P T Englund
Keyword(s):  
Mitochondrial Dna ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Kinetoplast Dna ◽  
Gap Filling ◽  
Synthesis And Processing ◽  
Trypanosoma Equiperdum

Kinetoplast DNA, the mitochondrial DNA in trypanosomes, is a giant network containing topologically interlocked minicircles. Replication occurs on free minicircles that have been detached from the network. In this paper, we report studies on the synthesis and processing of the minicircle L and H strands. Analysis of free minicircles from Trypanosoma equiperdum by two-dimensional agarose gel electrophoresis indicated that elongating L strands are present on theta structures. Hybridization studies indicated that L-strand elongation is continuous and unidirectional, starting near nucleotide 805 and proceeding around the entire minicircle. The theta structures segregate into monomeric progeny minicircles, and those with a newly synthesized L strand have a 8-nucleotide gap between nucleotides 805 and 814 (J. M. Ntambi, T. A. Shapiro, K. A. Ryan, and P. T. Englund, J. Biol. Chem. 261:11890-11895, 1986). These molecules are reattached to the network, where repair of the gap takes place. Of the molecules labeled during a 10-min pulse with [3H]thymidine, gap filling occurred on half within about 15 min and on virtually all by 60 min; however, there was no detectable covalent closure of the newly synthesized L strand by 60 min.

scholarly journals Synthesis and processing of kinetoplast DNA minicircles in Trypanosoma equiperdum.

1989 ◽  
Vol 9 (8) ◽  
pp. 3212-3217 ◽  
Author(s):  
K A Ryan ◽  
P T Englund
Keyword(s):  
Mitochondrial Dna ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Kinetoplast Dna ◽  
Gap Filling ◽  
Synthesis And Processing ◽  
Trypanosoma Equiperdum

Kinetoplast DNA, the mitochondrial DNA in trypanosomes, is a giant network containing topologically interlocked minicircles. Replication occurs on free minicircles that have been detached from the network. In this paper, we report studies on the synthesis and processing of the minicircle L and H strands. Analysis of free minicircles from Trypanosoma equiperdum by two-dimensional agarose gel electrophoresis indicated that elongating L strands are present on theta structures. Hybridization studies indicated that L-strand elongation is continuous and unidirectional, starting near nucleotide 805 and proceeding around the entire minicircle. The theta structures segregate into monomeric progeny minicircles, and those with a newly synthesized L strand have a 8-nucleotide gap between nucleotides 805 and 814 (J. M. Ntambi, T. A. Shapiro, K. A. Ryan, and P. T. Englund, J. Biol. Chem. 261:11890-11895, 1986). These molecules are reattached to the network, where repair of the gap takes place. Of the molecules labeled during a 10-min pulse with [3H]thymidine, gap filling occurred on half within about 15 min and on virtually all by 60 min; however, there was no detectable covalent closure of the newly synthesized L strand by 60 min.

A computer model for the analysis of DNA replication intermediates by two-dimensional agarose gel electrophoresis

Gene ◽  
1998 ◽  
Vol 217 (1-2) ◽  
pp. 41-49 ◽  
Author(s):  
E. Viguera ◽  
A. Rodrı́guez ◽  
P. Hernández ◽  
D.B. Krimer ◽  
O. Trellez ◽  
...  
Keyword(s):  
Dna Replication ◽  
Computer Model ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Replication Intermediates

THE NATURE OF TWO COMPONENTS OF PIG MUCOSAL HEPARIN, SEPARATED BY ELECTROPHORESIS IN AGAROSE GEL

1967 ◽  
Vol 45 (7) ◽  
pp. 1015-1020 ◽  
Author(s):  
Alan A. Horner
Keyword(s):  
Gel Electrophoresis ◽  
Nitrous Acid ◽  
Anticoagulant Activity ◽  
Thin Layers ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Uronic Acids ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis

Two components of pig mucosal heparin were separated by agarose-gel electrophoresis and recovered from the gels. The slower moving component had the higher anticoagulant activity and intrinsic viscosity. The faster moving component had the higher sulfate : carboxyl ratio. Each component was degraded by self-hydrolysis to N-desulfated heparin, deaminated with nitrous acid, and hydrolyzed with formic acid. Two-dimensional electrophoresis and chromatography on thin layers of cellulose showed that each component had two uronic acids with the characteristics of glucuronic and iduronic acids.

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