title: In-frame cDNA library combined with protein complementation assay identifies ARL11-binding partners.
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.
authors.affiliation: Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Sangkyou
authors.initials: S
authors.lastname: Lee
authors.forename: Ilkyun
authors.initials: I
authors.lastname: Lee
authors.forename: Yoonsuh
authors.initials: Y
authors.lastname: Jung
authors.forename: David
authors.initials: D
authors.lastname: McConkey
authors.forename: Bogdan
authors.initials: B
authors.lastname: Czerniak
mesh_terms: D020121:5' Untranslated Regions; D020727:ADP-Ribosylation Factors / Q000378:metabolism*; D001483:Base Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D002460:Cell Line; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D025941:Protein Interaction Mapping; D018168:Receptors, Retinoic Acid / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; D002352:Carrier Proteins; D018168:Receptors, Retinoic Acid; C082928:retinoic acid binding protein II, cellular; D020727:ADP-Ribosylation Factors; C500039:ARL11 protein, human