By John N. Abelson, Melvin I. Simon, Giuseppe M. Attardi, Anne Chomyn

The significantly acclaimed laboratory normal for 40 years, Methods in Enzymology is likely one of the so much hugely revered courses within the box of biochemistry. for the reason that 1955, each one quantity has been eagerlyawaited, often consulted, and praised via researchers and reviewers alike. greater than 260 volumes were released (all of them nonetheless in print) and masses of the cloth is suitable even today--truly a vital e-book for researchers in all fields of lifestyles sciences.

Key Features
* This quantity covers technique used in
* Mitochondrial gene expression
* Mitochondrial genetics and gene manipulation
* Mitochondrial illnesses and getting older

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Extra resources for Mitochondrial Biogenesis and Genetics Part B

Example text

Grivell, Curr. Genet. 23, 22 (1993). 7 T. D. Fox, Annu. Rev. Genet. 21, 67 (1987). s C. L. Dieckmann and R. R. Staples, Int. Rev. Cytol. 152, 145 (1994). METHODS IN ENZYMOLOGY, VOL. 264 Copyright © 1996 by Academic Press, Inc. All rights of reproduction in any form reserved. [4] PROTEIN AND R N A SYNTHESISSYSTEMSFROM YEAST 37 d e v e l o p e d 9-t2 for studying various aspects of m i t o c h o n d r i a l gene expression in vitro. T h e s e systems have b e e n used to d e t e r m i n e the stoichiometry of transcription and translation o f the m a j o r mitochondrial gene productsi°-t2; to d e t e r m i n e that m i t o c h o n d r i a l transcription and translation are n o t obligately coupledt2; to identify a p r e p r o t e i n p r e c u r s o r to c y t o c h r o m e - c oxidase subunit Ip3't4; to establish that mitochondrial translation p r o d u c t s are inserted into the inner m i t o c h o n d r i a l m e m b r a n e cotranslationallyIS'16; to examine the kinetic relationships b e t w e e n mitochondrial protein i m p o r t and m i t o c h o n d r i a l translationlT'lS; to e x a m i n e the assembly of respiratory proteins17'19'2°; and to study proteolytic t u r n o v e r of mitochondrial translation products.

Comparison of footprinting a m o n g different biochemically distinct stages of embryogenesis 5 is currently under investigation. Concluding Remarks The procedure described above yields reproducible, high-resolution of m t D N A templates. This should chondrial gene regulation in a wide is a simple footprinting protocol which genomic footprints from small amounts be generally applicable to study mitovariety of experimental samples where FIG. 3. The template in each reaction was mtDNA isolated from the mitochondria of X.

Biol. Chem. 266, 2333 (1991). z3 C. A. Virbasius, J. V. Virbasius, and R. C. Scarpulla, Genes Dev. 7, 2431 (1993). 24 G. S. Shadel and D. A. Clayton, J. Biol. Chem. 268, 16083 (1993). 25 A. Chomyn and G. Attardi, in "Molecular Mechanisms in Bioenergetics" (L. ), p. 483. Elsevier, Amsterdam, 1992. 125% DMS for 45 sec to obtain the naked D N A controls. The methylated and piperidine-cleaved D N A samples from the three preparations were used as template in P E F reactions, set up as described for a typical P E F reaction.

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