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- W2087644805 abstract "DT40 cells derived from chicken B lymphocytes exhibit exceptionally high homologous recombination rates. Therefore, they can be used as a convenient tool and model for gene targeting experiments. However, lack of efficient cloning strategies, protein purification protocols and a well annotated protein database limits the utility of these cells for proteomic studies. Here we describe a fast and inexpensive experimental pipeline for protein localization, quantification and mass spectrometry-based interaction studies using DT40 cells. Our newly designed set of pQuant vectors and a sequence- and ligation-independent cloning (SLIC) strategy allow for simple and efficient generation of gene targeting constructs, facilitating homologous-recombination-based protein tagging on a multi-gene scale. We also report proof of principle results using the key proteins involved in RNA decay, namely EXOSC8, EXOSC9, CNOT7 and UPF1." @default.
- W2087644805 created "2016-06-24" @default.
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- W2087644805 date "2013-07-27" @default.
- W2087644805 modified "2023-09-26" @default.
- W2087644805 title "A new strategy for gene targeting and functional proteomics using the DT40 cell line" @default.
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- W2087644805 doi "https://doi.org/10.1093/nar/gkt650" @default.
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