A pull-down procedure for the identification of unknown GEFs for small GTPases

用于识别小 GTPase 的未知 GEF 的下拉程序

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作者:Daniel Koch, Amrita Rai, Imtiaz Ali, Nathalie Bleimling, Timon Friese, Andreas Brockmeyer, Petra Janning, Bruno Goud, Aymelt Itzen, Matthias P Müller, Roger S Goody

Procedures

An in vitro transcription/translation system coupled with a GEF activity assay and a yeast two-hybrid screen for detection of GEFs. Although the procedures were established and tested using the Rab protein Sec4, the similar basic principle of action of all nucleotide exchange factors will allow the method to be used for identification of unknown GEFs of small GTPases in general.

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