Expression and purification of mitochondrial RNA polymerase and transcription factor a from drosophila melanogaster

John P. Gajewski, Jamie Jon Arnold, Tiina S. Salminen, Laurie S. Kaguni, Craig Eugene Cameron

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Mitochondrial gene expression is essential in all organisms. Our understanding of mitochondrial transcription on a biochemical level has been limited by the inability to purify the individual protein components involved in mitochondrial gene expression. Recently, new systems have been identified that permit purification of these proteins from bacteria. However, the generalizability of these systems is not clear. Here, we have applied the technology from the Cameron lab to express and purify mitochondrial RNA polymerase and transcription factor A from Drosophila melanogaster . We show that the use of SUMO system to produce SUMO fusion proteins in bacteria is effective not only for the human and mouse proteins, but also for the fl y proteins. The application of this system to produce the mitochondrial proteins from other organisms should permit detailed understanding of mitochondrial transcription from any organism.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages199-210
Number of pages12
DOIs
StatePublished - Jan 1 2016

Publication series

NameMethods in Molecular Biology
Volume1351
ISSN (Print)1064-3745

Fingerprint

DNA-Directed RNA Polymerases
Drosophila melanogaster
Transcription Factors
Mitochondrial Genes
Small Ubiquitin-Related Modifier Proteins
Proteins
Bacteria
Gene Expression
Mitochondrial Proteins
Technology
mitochondrial RNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Gajewski, J. P., Arnold, J. J., Salminen, T. S., Kaguni, L. S., & Cameron, C. E. (2016). Expression and purification of mitochondrial RNA polymerase and transcription factor a from drosophila melanogaster. In Methods in Molecular Biology (pp. 199-210). (Methods in Molecular Biology; Vol. 1351). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3040-1_15
Gajewski, John P. ; Arnold, Jamie Jon ; Salminen, Tiina S. ; Kaguni, Laurie S. ; Cameron, Craig Eugene. / Expression and purification of mitochondrial RNA polymerase and transcription factor a from drosophila melanogaster. Methods in Molecular Biology. Humana Press Inc., 2016. pp. 199-210 (Methods in Molecular Biology).
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Gajewski, JP, Arnold, JJ, Salminen, TS, Kaguni, LS & Cameron, CE 2016, Expression and purification of mitochondrial RNA polymerase and transcription factor a from drosophila melanogaster. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1351, Humana Press Inc., pp. 199-210. https://doi.org/10.1007/978-1-4939-3040-1_15

Expression and purification of mitochondrial RNA polymerase and transcription factor a from drosophila melanogaster. / Gajewski, John P.; Arnold, Jamie Jon; Salminen, Tiina S.; Kaguni, Laurie S.; Cameron, Craig Eugene.

Methods in Molecular Biology. Humana Press Inc., 2016. p. 199-210 (Methods in Molecular Biology; Vol. 1351).

Research output: Chapter in Book/Report/Conference proceedingChapter

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Gajewski JP, Arnold JJ, Salminen TS, Kaguni LS, Cameron CE. Expression and purification of mitochondrial RNA polymerase and transcription factor a from drosophila melanogaster. In Methods in Molecular Biology. Humana Press Inc. 2016. p. 199-210. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-3040-1_15