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High‐throughput generation of tagged stable cell lines for proteomic analysis
Author(s) -
Torres Jorge Z.,
Miller Julie J.,
Jackson Peter K.
Publication year - 2009
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800873
Subject(s) - computational biology , proteomics , microbiology and biotechnology , microtubule , cell culture , identification (biology) , biology , flexibility (engineering) , throughput , cell cycle , cell , chemistry , biochemistry , computer science , genetics , gene , telecommunications , statistics , botany , mathematics , wireless
We present an optimized system for rapid generation of localization and affinity purification‐tagged mammalian stable cell lines that facilitates complex purification and interacting protein identification. The improved components of this method, including the flexibility of inducible expression, circumvent issues associated with toxicity, clonal selection, sample yields and time to data acquisition. We have applied this method to the study of cell‐cycle regulators and novel microtubule‐associated proteins.