Abstract
Kinesin and kinesin-related proteins comprise a family of molecular motors that utilize the chemical energy provided by the hydrolysis of AT P to perform force-generating movements along filamentous microtubules. Members of this family of microtubule motors are vital for numerous cellular activities, such as organelle transport and chromosome segregation. Kinesin-related proteins are not only linked by function, but each family member shares a similar “motordomain” region responsible for converting the chemical energy of ATP into mechanical force (reviewed in refs. 1 and 2). Within this specialized motor region, there exist short, conserved sequences that are not only present in kinesin-related motors but also found in other ATP-hydrolyzing molecular motors (myosins and dyneins) and GTP-hydrolyzing G proteins (reviewed in ref. 3). In this chapter, we will describe our use of pan-specific, kinesin peptide antibodies generated against two of these short, conserved sequences for the expression cloning of members of the kinesin-related protein family. We will briefly discuss the production of the pan-specific, kinesin peptide antisera and the four conserved regions within the motor domain of the kinesin protein family previously selected for the generation of these antibodies. In more detail, we will describe the method by which our laboratory has used these pan-specific antibodies to identify and clone kinesin-related proteins.
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Ginkel, L.M., Wordeman, L. (2001). Expression Cloning with Pan Kinesin Antibodies. In: Vernos, I. (eds) Kinesin Protocols. Methods in Molecular Biology™, vol 164. Humana Press. https://doi.org/10.1385/1-59259-069-1:21
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DOI: https://doi.org/10.1385/1-59259-069-1:21
Publisher Name: Humana Press
Print ISBN: 978-0-89603-766-3
Online ISBN: 978-1-59259-069-8
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