In-Frame cDNA Library Combined with Protein Complementation Assay Identifies ARL11-Binding Partners |
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| Authors: | Sangkyou Lee Ilkyun Lee Yoonsuh Jung David McConkey Bogdan Czerniak |
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| Institution: | 1. Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.; 2. Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.; 3. Department of Urology, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.; Colorado State University, United States of America, |
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| Abstract: | The cDNA expression libraries that produce correct proteins are essential in facilitating the identification of protein-protein interactions. The 5′-untranslated regions (UTRs) that are present in the majority of mammalian and non-mammalian genes are predicted to alter the expression of correct proteins from cDNA libraries. We developed a novel cDNA expression library from which 5′-UTRs were removed using a mixture of polymerase chain reaction primers that complement the Kozak sequences we refer to as an “in-frame cDNA library.” We used this library with the protein complementation assay to identify two novel binding partners for ras-related ADP-ribosylation factor-like 11 (ARL11), cellular retinoic acid binding protein 2 (CRABP2), and phosphoglycerate mutase 1 (PGAM1). Thus, the in-frame cDNA library without 5′-UTRs we describe here increases the chance of correctly identifying protein interactions and will have wide applications in both mammalian and non-mammalian detection systems. |
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