A generic tool for biotinylation of tagged proteins in transgenic mice.
作者： Siska Driegen , Rita Ferreira , Arend van Zon , John Strouboulis , Martine Jaegle
DOI: 10.1007/S11248-005-7220-2
关键词: Transgene 、 Biology 、 Protein tag 、 Biochemistry 、 Transfection 、 Genetically modified mouse 、 Avidin 、 Streptavidin 、 Biotinylation 、 Biotin
摘要: The remarkable high affinity (Kd ~ 10−15 M) of avidin/streptavidin for biotin has been extensively exploited in purification methodologies. Recently a small peptide sequence (Avi-tag) defined that can be specifically and efficiently biotinylated by the bacterial BirA ligase. Fusion this to protein interest co-expression with gene mammalian cells allowed together its associated proteins other molecules. Ideally, one would like apply these technologies purify tagged directly from mouse tissues. To make approach feasible large variety we developed strain expresses ubiquitously inserting it ROSA26 locus. We demonstrate is indeed expressed all tissues tested. In order functionality show biotinylates transgene-encoded Avi-tagged Gata1 Oct6 transcription factors erythroid foetal liver Schwann peripheral nerve respectively. Therefore, crossed any transgenic obtain efficient biotinylation an purpose (complex) purification.
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