Document Detail


High-throughput cloning and expression library creation for functional proteomics.
MedLine Citation:
PMID:  23457047     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The study of protein function usually requires the use of a cloned version of the gene for protein expression and functional assays. This strategy is particularly important when the information available regarding function is limited. The functional characterization of the thousands of newly identified proteins revealed by genomics requires faster methods than traditional single-gene experiments, creating the need for fast, flexible, and reliable cloning systems. These collections of ORF clones can be coupled with high-throughput proteomics platforms, such as protein microarrays and cell-based assays, to answer biological questions. In this tutorial, we provide the background for DNA cloning, discuss the major high-throughput cloning systems (Gateway® Technology, Flexi® Vector Systems, and Creator(TM) DNA Cloning System) and compare them side-by-side. We also report an example of high-throughput cloning study and its application in functional proteomics. This tutorial is part of the International Proteomics Tutorial Programme (IPTP12).
Authors:
Fernanda Festa; Jason Steel; Xiaofang Bian; Joshua Labaer
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Review     Date:  2013-04-05
Journal Detail:
Title:  Proteomics     Volume:  13     ISSN:  1615-9861     ISO Abbreviation:  Proteomics     Publication Date:  2013 May 
Date Detail:
Created Date:  2013-04-22     Completed Date:  2013-12-10     Revised Date:  2014-04-25    
Medline Journal Info:
Nlm Unique ID:  101092707     Medline TA:  Proteomics     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1381-99     Citation Subset:  IM    
Copyright Information:
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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MeSH Terms
Descriptor/Qualifier:
Bacteriophage lambda
Cell-Free System
Cloning, Molecular / methods*
DNA Restriction Enzymes / metabolism
Enzymes / genetics,  metabolism
Gene Expression
Gene Library
Humans
Open Reading Frames
Promoter Regions, Genetic
Proteomics / methods*
Recombination, Genetic
Vibrio cholerae / genetics
Grant Support
ID/Acronym/Agency:
U01 AI077883/AI/NIAID NIH HHS; U01AI077883/AI/NIAID NIH HHS; U01CA117374/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Enzymes; EC 3.1.21.-/DNA Restriction Enzymes

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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