Protein-trap version 2.1: screening for expressed proteins in mammalian cells based on their localizations.
BACKGROUND
"Protein-trap" is a method that allows epitope-tagging of endogenous proteins. This method allows for the identification of endogenously expressed proteins that exhibit specific localization of interest. This method has been recently reported for its application in the study of Drosophila development by using a relatively large epitope, green-fluorescent-protein (GFP).


RESULTS
Herein, we report a new "protein-trap" vector for mammalian cells. This new method utilizes a much smaller epitope-tag and also allows for drug-selection prior to the epitope-tagging. Pre-selection by drug resulted in the highly efficient protein-trapping frequency.


CONCLUSIONS
The "protein-trap" method based on this new vector is expected to serve as a complimentary approach to the previously reported GFP-based method.
Sineshchekova
Olga O
OO
The University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390, USA. Olga.Sineshchekova@UTSouthwestern.edu
Kawate
Toshimitsu
T
Vdovychenko
Oleksandr V
OV
Sato
Thomas N
TN
D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002277:Carcinoma / Q000737:chemistry / Q000235:genetics / Q000473:pathology / Q000821:virology; D003001:Cloning, Molecular / Q000379:methods*; D000939:Epitopes / Q000096:biosynthesis / Q000235:genetics; D005091:Exons / Q000235:genetics / Q000276:immunology; D005109:Extracellular Matrix / Q000502:physiology; D005822:Genetic Vectors / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D042541:Intracellular Space / Q000737:chemistry; D008175:Lung Neoplasms / Q000737:chemistry / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D014176:Protein Biosynthesis; D021381:Protein Transport / Q000502:physiology; D011506:Proteins / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D015398:Signal Transduction / Q000502:physiology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011506:Proteins; D011993:Recombinant Fusion Proteins