Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1977 Jul;60(1):98–101. doi: 10.1104/pp.60.1.98

DNA Binding and Uptake by Nuclei Isolated from Plant Protoplasts

Factors Affecting DNA Binding and Uptake 1

Kanji Ohyama a, Lawrence E Pelcher a, Dave Horn a
PMCID: PMC542556  PMID: 16660053

Abstract

DNA binding and uptake by nuclei isolated from soybean (Glycine max L. Merr.) protoplasts were investigated using radioactive homogeneous DNA prepared from soybean cells. DNA binding to nuclei was found to decrease drastically with increased incubation time. Total uptake and acid-precipitable uptake reached a maximum after 20 minutes of incubation. Optimum DNA binding and uptake occurred at pH 6 and the process was enhanced by increasing the incubation temperature to 40 C. Salmonella typhimurium DNA and poly ([dA-dT]-[dA-dT]) competitively inhibited DNA binding whereas calf thymus DNA was less competitive; however, Micrococcus lysodeikticus DNA stimulated DNA binding and tobacco mosaic virus RNA had no effect. DNA binding and uptake was enhanced by addition of Mg ions, Ca ions, poly-l-lysine, and ATP. Increasing amounts of EDTA appeared to decrease DNA binding. Pronase strongly inhibited DNA binding and uptake.

Full text

PDF
99

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aoki S., Takebe I. Infection of tobacco mesophyll protoplasts by tobacco mosaic virus ribonucleic acid. Virology. 1969 Nov;39(3):439–448. doi: 10.1016/0042-6822(69)90092-0. [DOI] [PubMed] [Google Scholar]
  2. Cosloy S. D., Oishi M. Genetic transformation in Escherichia coli K12. Proc Natl Acad Sci U S A. 1973 Jan;70(1):84–87. doi: 10.1073/pnas.70.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gamborg O. L. The effects of amino acids and ammonium on the growth of plant cells in suspension culture. Plant Physiol. 1970 Apr;45(4):372–375. doi: 10.1104/pp.45.4.372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ledoux L., Huart R., Jacobs M. DNA-mediated genetic correction of thiamineless Arabidopsis thaliana. Nature. 1974 May 3;249(452):17–21. doi: 10.1038/249017a0. [DOI] [PubMed] [Google Scholar]
  5. Ohyama K., Gamborg O. L., Miller R. A. Isolation and Properties of Deoxyribonucleic Acid from Protoplasts of Cell Suspension Cultures of Ammi visnaga and Carrot (Daucus carota). Plant Physiol. 1972 Sep;50(3):319–321. doi: 10.1104/pp.50.3.319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Seto H., Tomasz A. Early stages in DNA binding and uptake during genetic transformation of pneumococci. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1493–1498. doi: 10.1073/pnas.71.4.1493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Uchimiya H., Murashige T. Quantitative analysis of the fate of exogenous DNA in Nicotiana protoplasts. Plant Physiol. 1977 Feb;59(2):301–308. doi: 10.1104/pp.59.2.301. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES