Skip to main content
Log in

Characterization of 3H-labeled supercoiled mitochondrial DNA from tobacco suspension culture cells

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Mitochondrial DNA of tobacco supension culture cells labeled in vivo with 3H-thymidine was isolated from purified mitochondria. It has a density of 1.707±0.002 g/cm3 unique from both nuclear and chloroplast DNA. Approximately 25–45% of the mtDNA molecules from several preparations from suspension culture cells were isolated as supercoiled molecules from ethidium bromide-cesium chloride gradients. Electron microscopic examination of nicked supercoiled molecules revealed a heterogenous population of molecules ranging in size from 0.5 to 42 μm. The most abundant size class of 365 molecules measured was 9 μm. Restriction endonuclease digests of both supercoiled and open circular/linear molecules were essentially identical and yielded a minimum size estimate of 76 μm. The availability of large amounts of labeled covalently closed circular molecules will facilitate the resolution of the different size classes and allow, for the first time, a direct analysis of the relationships among the heterogenous population of DNA molecules found in plant mitochondria.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bogenhagen D, Clayton DA (1977) Mouse L cell mitochondrial DNA molecules are selected randomly for replication throughout the cell cycle. Cell 2:719–727

    Google Scholar 

  • Borst P (1972) Mitochondrial nucleic acids. Annu Rev Biochem 41:333–376

    Google Scholar 

  • Cummings DJ, Belcour L, Grandchamp C (1979) Mitochondrial DNA from Podospora Anserina II. Properties of mutant DNA and multimeric circular DNA from senescent DNA. Mol Gen Genet 171:239–250

    Google Scholar 

  • Ferguson J, Davis RW (1978) Quantitative electron microscopy of nucleic acids. In: Kochler (ed) Advanced techniques in biological electron microscopy, vol 2. Spring Verlag, Berlin p 123

    Google Scholar 

  • Kolodner R, Tewari KK (1972) Physicochemical characterization of mitochondrial DNA from pea leaves. Proc Natl Acad Sci USA 69:1830–1834

    Google Scholar 

  • Levings CS, Pring DR (1979) Organization and information content of mitochondrial DNA of higher plants. J Supramol Struct 3:134–137

    Google Scholar 

  • Levings CS, Pring DR (1977) Diversity of mitochondrial genomes among normal cytoplasms of maize. J Hered 68:350–354

    Google Scholar 

  • Levings CS, Pring DR (1976) Restriction endonuclease analysis of mitochondrial DNA from normal and Texas male-sterile cytoplasms of maize. Science 193:158–160

    Google Scholar 

  • Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol plant 18:100–127

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory New York

    Google Scholar 

  • Quetier F, Vedel F (1977) Heterogeneous population of mitochondrial DNA molecules in higher plants. Nature 268:365–368

    Google Scholar 

  • Schildkraut CL, Marmur J, Doty P (1962) Determination of the base composition of DNA from its buoyant density in CsC1. J Mol Biol 4:430–433

    Google Scholar 

  • Syrenki RM, Levings CS, Shah DM (1978) Physicochemical characterization of mitochondrial DNA from soybean. Plant Physiol 61:460–464

    Google Scholar 

  • Wong Flossie Y, Wildman SG (1972) Simple procedure for isolation of satellite DNA's from tobacco leaves in high yield and demonstration of minicireles. Biochim Biophys Acta 259:5–12

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by L.S. Lerman

Dr. R. Dale and part of this research was supported by NIH Grant GM 24775

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sparks, R.B., Dale, R.M.K. Characterization of 3H-labeled supercoiled mitochondrial DNA from tobacco suspension culture cells. Molec. Gen. Genet. 180, 351–355 (1980). https://doi.org/10.1007/BF00425847

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00425847

Keywords

Navigation