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Systematic subcellular localization of novel proteins identified by large‐scale cDNA sequencing
Author(s) -
Simpson Jeremy C,
Wellenreuther Ruth,
Poustka Annemarie,
Pepperkok Rainer,
Wiemann Stefan
Publication year - 2000
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1093/embo-reports/kvd058
Subject(s) - subcellular localization , biology , protein subcellular localization prediction , computational biology , complementary dna , cloning (programming) , fusion protein , genome , gene , cdna library , genetics , microbiology and biotechnology , computer science , recombinant dna , programming language
As a first step towards a more comprehensive functional characterization of cDNAs than bioinformatic analysis, which can only make functional predictions for about half of the cDNAs sequenced, we have developed and tested a strategy that allows their systematic and fast subcellular localization. We have used a novel cloning technology to rapidly generate N‐ and C‐terminal green fluorescent protein fusions of cDNAs to examine the intracellular localizations of >100 expressed fusion proteins in living cells. The entire analysis is suitable for automation, which will be important for scaling up throughput. For >80% of these new proteins a clear intracellular localization to known structures or organelles could be determined. For the cDNAs where bioinformatic analyses were able to predict possible identities, the localization was able to support these predictions in 75% of cases. For those cDNAs where no homologies could be predicted, the localization data represent the first information.