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Single-Molecule Analysis of Sf9 Purified Superprocessive Kinesin-3 Family Motors
JoVE 杂志
生物化学
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JoVE 杂志 生物化学
Single-Molecule Analysis of Sf9 Purified Superprocessive Kinesin-3 Family Motors

Single-Molecule Analysis of Sf9 Purified Superprocessive Kinesin-3 Family Motors

1,720 Views

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08:16 min

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July 27, 2022

DOI:

10.3791/63837-v

DOI:
10.3791/63837-v

•

08:16 min
July 27, 2022

•

2 Views
, , ,
1Discipline of Biological Engineering,Indian Institute of Technology Gandhinagar, 2Department of Biotechnology and Bioinformatics,Sambalpur University

Summary

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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.

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Sf9 CellsBaculovirus ExpressionSingle-molecule AnalysisKinesin-3 MotorsSuperprocessive MotorsSingle-molecule MotilityMulti-motor Gliding AssaysCytoskeletal ProteinsDionineMyosinProtein PurificationAffinity Purification

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Cite this Article

Soppina, P., Shewale, D. J., Naik, P. K., Soppina, V. Single-Molecule Analysis of Sf9 Purified Superprocessive Kinesin-3 Family Motors. J. Vis. Exp. (185), e63837, doi:10.3791/63837 (2022).

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