DNA unwinding assay using streptavidin-bound oligonucleotides
2006
Improved Method for Determining Helicase Directionality
publication
Evidence: moderate
Author Information
Author(s): Shin Jae-Ho, Kelman Zvi
Primary Institution: University of Maryland Biotechnology Institute
Hypothesis
Can biotin-streptavidin complexes improve the determination of helicase directionality?
Conclusion
The use of a biotin-streptavidin complex as a helicase substrate improves helicase activity and the determination of helicase directionality.
Supporting Evidence
- The new assay improves helicase activity for several DNA helicases.
- A universal substrate was developed to determine the directionality of both 3'→ 5' and 5'→ 3' helicases.
- Streptavidin-bound substrates allow for easy determination of helicase directionality.
Takeaway
This study shows that using special biotin-labeled strands helps scientists figure out which way certain enzymes move when they unwind DNA.
Methodology
The study developed an improved assay using biotinylated oligonucleotides to determine helicase directionality.
Limitations
Some helicases may displace streptavidin from biotinylated oligonucleotides, which could affect the results.
Digital Object Identifier (DOI)
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