In Vitro Reconstitution of Human Mitochondrial Transcription

In vitro assay based on a reconstituted mitochondrial transcription system serves as a method of choice to probe the functional importance of proteins and their structural motifs. Here we describe protocols for transcription assays designed to probe activity of the human mitochondrial RNA polymerase...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2192; p. 35
Main Authors Sarfallah, Azadeh, Temiakov, Dmitry
Format Journal Article
LanguageEnglish
Published United States 2021
Subjects
DNA, Mitochondrial - chemistry
DNA, Mitochondrial - genetics
DNA-Binding Proteins - chemistry
DNA-Directed RNA Polymerases - chemistry
Escherichia coli - genetics
Escherichia coli - metabolism
Humans
In Vitro Techniques
Methyltransferases - chemistry
Mitochondria - metabolism
Mitochondrial Proteins - chemistry
Promoter Regions, Genetic
Protein Binding
Transcription Factors - chemistry
Transcription, Genetic
Promoter
TFAM
TFB2M
RNAP
POLRTM
Mitochondrial transcription
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Summary:In vitro assay based on a reconstituted mitochondrial transcription system serves as a method of choice to probe the functional importance of proteins and their structural motifs. Here we describe protocols for transcription assays designed to probe activity of the human mitochondrial RNA polymerase and the transcription initiation complex using RNA-DNA scaffold and synthetic promoter templates.
ISSN:1940-6029
DOI:10.1007/978-1-0716-0834-0_3