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A novel method for purification of poly(ADP-ribose)
Journal of Biochemistry
|September 1, 1980
Summary
Researchers developed a new method to purify poly(ADP-ribose) (PAR). This technique efficiently isolates PAR synthesized in calf thymus nuclei, yielding a pure product suitable for further analysis.
Area of Science:
- Biochemistry
- Molecular Biology
Background:
- Poly(ADP-ribose) (PAR) plays crucial roles in DNA repair and gene regulation.
- Efficient purification of PAR is essential for studying its biological functions.
Purpose of the Study:
- To establish a novel and efficient method for the purification of poly(ADP-ribose) (PAR).
Main Methods:
- Incubation of calf thymus nuclei with NAD.
- Pronase digestion and DNA isolation using NaCl and ethanol precipitation.
- Nuclease treatment followed by hydroxylapatite column chromatography for PAR purification.
Main Results:
- Acid-insoluble PAR synthesized during incubation was quantitatively recovered in the DNA fraction.
- Purification yielded PAR as a single peak on a hydroxylapatite column.
- The purified PAR exhibited characteristics of purity, including an A280/A260 ratio of 0.24.
Conclusions:
- A novel, effective method for poly(ADP-ribose) purification has been successfully established.
- This method allows for the quantitative recovery and high-purity isolation of PAR.
- The purified PAR is suitable for biochemical and molecular analyses.