Expression Cloning with Pan Kinesin Antibodies
| Abstract |
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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.
Affiliation(s): (2) Department of Physiology and Biophysics, University of Washington School of Medicine, Seattle, WA
(3) Department of Physiology and Biophysics, University of Washington School of Medicine, Seattle, WA
(3) Department of Physiology and Biophysics, University of Washington School of Medicine, Seattle, WA
Book Title: Kinesin Protocols
Series: Methods in Molecular Biology | Volume: 164 | Pub. Date: Dec-03-2000 | Page Range: 21-41 | DOI: 10.1385/1-59259-069-1:21
Subject: Protein Science
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