Article thumbnail

Identification of RNA-Binding Proteins by RNA Ligand-based cDNA Expression Library Screening

By ?????????

Abstract

We previously reported when a portion of the Requiem (REQ/DPF2) messenger ribonucleic acid (mRNA) 3’ untranslated region (3’UTR), referred to as G8, was overexpressed in K562 cells, β-globin expression was induced, suggesting that the 3’UTR of REQ mRNA plays a physiological role (Kim et al., 2014). To identify trans-acting factors that bind to the REQ 3’UTR, we describe the RNA ligand based cDNA expression library screening method. This protocol could be adapted to detect specific RNA-protein interactions. Following this method, we identified six positive clones in the initial round of screening and four pure clones after sib-screening. This protocol was originally published in Kim et al. (2014).This work was supported by Basic Science Research Program (2010-00252250 to C. G. K.), National Research Foundation (NRF), Ministry of Education, Science and Technology (MEST), Republic of Korea. Converging Research Center Program (2013K000283 to C. G. K.), Ministry of Science, ICT & Future Planning (MSIFP), Republic of Korea

Topics: RNA-protein interaction, Phage display, Requiem (REQ/DPF2), CDNA library screening
Publisher: Bio-protocol LLC
Year: 2016
DOI identifier: 10.21769/bioprotoc.1715
OAI identifier: oai:repository.hanyang.ac.kr:20.500.11754/33996
Provided by: HANYANG Repository
Journal:
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://hdl.handle.net/20.500.1... (external link)
  • http://www.bio-protocol.org/e1... (external link)
  • Suggested articles


    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.