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Publication : Cloning and deletion mutagenesis using direct protein-protein interaction on an expression vector. Identification of the calmodulin binding domain of alpha-fodrin.

First Author  Sri Widada J Year  1989
Journal  J Mol Biol Volume  205
Issue  2 Pages  455-8
PubMed ID  2926814 Mgi Jnum  J:43398
Mgi Id  MGI:1097596 Doi  10.1016/0022-2836(89)90355-0
Citation  Sri Widada J, et al. (1989) Cloning and deletion mutagenesis using direct protein-protein interaction on an expression vector. Identification of the calmodulin binding domain of alpha-fodrin. J Mol Biol 205(2):455-8
abstractText  We have screened a lambda gt11 library, constructed with mouse macrophage cDNA, in order to isolate clones that code for calmodulin binding proteins. We have developed a new approach for this purpose using radioactive calmodulin (produced by genetic engineering) to detect fusion proteins that interact with this protein with high affinity. A cDNA clone that codes for mouse macrophage fodrin was isolated, sequenced and identified. By deleting part of the sequence the calmodulin binding domain was located on the fodrin sequence. The site is situated on repeat 11 of fodrin and probably on the extra arm of this repeat. The method we developed is widely applicable to site-directed mutagenesis of interacting proteins.
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