In-Frame cDNA Library Combined with Protein Complementation Assay Identifies ARL11-Binding Partners

Sangkyou Lee, Ilkyun Lee, Yoonsuh Jung, David McConkey, Bogdan Czerniak

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Article numbere52290
JournalPloS one
Volume7
Issue number12
DOIs
Publication statusPublished - 2012 Dec 18
Externally publishedYes

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ADP-Ribosylation Factors
5' untranslated regions
Gene Library
cDNA libraries
Assays
5' Untranslated Regions
assays
phosphoglyceric acids
Proteins
proteins
retinoic acid
protein-protein interactions
binding proteins
complement
Complementary DNA
Phosphoglycerate Mutase
protein synthesis
polymerase chain reaction
Retinoic Acid Receptors
Polymerase chain reaction

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

In-Frame cDNA Library Combined with Protein Complementation Assay Identifies ARL11-Binding Partners. / Lee, Sangkyou; Lee, Ilkyun; Jung, Yoonsuh; McConkey, David; Czerniak, Bogdan.

In: PloS one, Vol. 7, No. 12, e52290, 18.12.2012.

Research output: Contribution to journalArticle

Lee, Sangkyou ; Lee, Ilkyun ; Jung, Yoonsuh ; McConkey, David ; Czerniak, Bogdan. / In-Frame cDNA Library Combined with Protein Complementation Assay Identifies ARL11-Binding Partners. In: PloS one. 2012 ; Vol. 7, No. 12.
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