P1 Virus Stock Generation and Large-Scale Infection
3:28
Sf9 Purification of Kinesin-3 Motors
4:25
Microtubule Polymerization and In Vitro Single-Molecule Motility Assay
5:22
In Vitro Microtubule Gliding Assay
6:23
Results: Analysis of Sf9-Purified Kinesin-3 Motor
7:24
Conclusion
Transcript
The protocol offers a simple method for purifying active molecular motors using Sf9 baculovirus system. It also details single molecule motility and multi-motor gliding assays to study the regulatory mechanism of motor proteins. All that tradition
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This study details purification of KIF1A(1-393LZ), a member of kinesin-3 family, using Sf9-baculovirus expression system. In vitro single-molecule and multi-motor gliding analysis of these purified motors exhibited robust motility properties comparable to motors from mammalian cell lysate. Thus, Sf9-baculovirus system is amenable to express and purify motor protein of interest.