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. Author manuscript; available in PMC: 2012 Sep 15.
Published in final edited form as: J Neurosci Methods. 2011 Jul 2;200(2):121–128. doi: 10.1016/j.jneumeth.2011.06.020

Figure 1.

Figure 1

Experimental designs for cloning, expression and purification of mBtNGF. As described in Materials and Methods, a mouse cDNA sequence encoding the full length of prepro NGF with a C-terminal AviTag is co-transfected with an expression vector for the E.coli biotin ligase (BirA) into 293FT cells. The specific “Lysine” residue within the AviTag is ligated to a biotin by BirA. Upon or during secretion, the N-terminal prepro sequence is cleaved and the mature protein with the biotinylated AviTag is released into the media. Mono-biotinylated NGF is secreted into the media and can be recovered from the media by Ni-NTA purification.