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Volume 99, Issue 2

Isolation of Mitochondria and Mitochondrial RNA from Free

Abstract

Summary: Two methods were used to isolate mitochondria from In the first method, cells were weakened by exposure to hypotonic conditions and then disrupted by blending; mitochondria were subsequently isolated using disodium 3,5-diacetoamido-2,4,6-triiodobenzoate gradients. In the second, cells were treated with digitonin before disruption; mitochondria were purified by differential centrifugation. Both preparations were examined with the electron microscope and were also shown to possess several characteristic biochemical properties of mitochondria. Kinetoplast DNA was present in the mitochondria, uncontaminated by nuclear DNA.

Analysis by polyacrylamide gel electrophoresis showed two RNA components of molecular weights 0·47 × 10 and 0·22 × 10, in addition to cytoplasmic RNA contamination. Four mitochondrial components with sedimentation coefficients of 14·6S, 11·4S, 10·1S and 6·9S were identified on sucrose density gradients. Ethidium bromide abolished the incorporation of [5-H] uridine into the presumed mitochondrial RNA.

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© Society for General Microbiology 1977
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1977-04-01
2024-04-19
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Isolation of Mitochondria and Mitochondrial RNA from Crithidia fasciculata
Microbiology 99, 291 (1977); https://doi.org/10.1099/00221287-99-2-291
/content/journal/micro/10.1099/00221287-99-2-291
/content/journal/micro/10.1099/00221287-99-2-291
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