Mitochondrial DNA I. Preparation and properties of mitochondrial DNA from chick liver

Borst, Piet; Ruttenberg, G.J.C.M.; Kroon, A.M.

doi:10.1016/0005-2787(67)90697-1

Cited by 113 publications

(6 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CEF were prepared from 8-to 9-day-old White Leghorn embryos (Spafas) and passaged as described previously (20). The basic medium for culturing the cells was Ham F12 supplemented with 8% calf serum, 2% inactivated chicken serum, and 4 ,ug of uridine per ml. Penicillin (100 IU/ml), streptomycin (100 ,ug/ml), and fungizone (0.5 ,ug/ml) were routinely added to the culture medium.…”

Section: Methodsmentioning

confidence: 99%

Ethidium bromide-induced loss of mitochondrial DNA from primary chicken embryo fibroblasts.

Desjardins¹,

Frost²,

Morais³

1985

Mol. Cell. Biol.

156

View full text Add to dashboard Cite

Chicken embryo fibroblasts in uridine-containing medium are inherently resistant to the growth-inhibitory effect of ethidium bromide. The drug was found to inhibit the incorporation of [3lHlthymidine into mitochondrial DNA circular molecules. Mitochondrial DNA was quantitated by DNA-DNA reassociation kinetics with a probe of chicken liver mitochondrial DNA. A mean number of 604 copies of mitochondrial DNA per cell was found. This number decreased progressively in cells exposed to ethidium bromide, and by day 13 ca. one copy of mitochondrial DNA was detected per cell. When the cells were then transferred to drug-free medium, the number of copies increased very slowly as a function of time. On the other hand, analyses of DNA extracted from cell populations exposed to ethidium bromide for 20 or more days, with or without subsequent transfer to drug-free medium, revealed very little or no mitochondrial DNA by reassociation kinetics or by Southern blot hybridization of AvaI-or HindIII-digested total cellular DNA. As a result of the elimination of mitochondrial DNA molecules, the establishment of cell populations with a respiration-deficient phenotype was confirmed by measuring cytochrome c oxidase activity as a function of the number of cell generations and the absorption spectrum of mitochondrial cytochromes.

show abstract

Section: Methodsmentioning

confidence: 99%

Ethidium bromide-induced loss of mitochondrial DNA from primary chicken embryo fibroblasts.

Desjardins¹,

Frost²,

Morais³

1985

Mol. Cell. Biol.

156

View full text Add to dashboard Cite

show abstract

“…Six livers were pooled, cut into small peices with scissors and homogenized with a glass-teflon Potter Elvehjem homogenizer using 0.25 M sucrose as the medium. Mitochondria were prepared by differential centrifugation as described previously [4]. The final mitochondrial pellet was resuspended to a volume of 12 ml in a medium containing 50 mM Tris buffer, 10 mM NaCl and 10 mM EDTA, pH 7.5.…”

Section: Preparation Of Mitochondrial Dna From Rat Livermentioning

confidence: 99%