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Characterization of plant polyadenylation complexes by using tandem affinity purification.
MedLine Citation:
PMID:  25487205     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Messenger RNA in eukaryotic cells is initially produced as a nascent transcript (pre-mRNA) without a polyadenine [poly(A)] tail to the 3' end. The precise cleavage of the pre-mRNA and addition of a poly(A) track need the communication between cis-elements in the pre-mRNA sequences and transacting protein factors recognizing them. Based on homology analyses, Arabidopsis cleavage and polyadenylation specificity factor (AtCPSF) complex should play a critical role in pre-mRNA 3' end processing. Here we describe the isolation of AtCPSF complex by using a tandem affinity purification (TAP) method. We demonstrate that TAP is a potent protein complex isolating approach that can fulfill a downstream protein identification purpose based on mass spectrometry techniques.
Authors:
Hongwei Zhao; Xinfu Ye; Qingshun Quinn Li
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  1255     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2015  
Date Detail:
Created Date:  2014-12-09     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  69-78     Citation Subset:  IM    
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