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. 2009 Jul 17;37(18):e122. doi: 10.1093/nar/gkp589

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© 2009 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — In vitro expression tests measuring the activity of Avi-AGT following active site dependent labelling of AGT with BG-biotin. DNA templates were either amplified from assembly reactions or plasmid DNA. L1, no template control; L2, biotinylated protein marker; L3, amplified with Taq from plasmid DNA; L4, amplified with Taq from an assembly reaction; L5, amplified with PfuTurbo C_x from plasmid DNA; L6, amplified with PfuTurbo C_x from an assembly reaction. The band at ∼20 kDa corresponds to biotin carboxyl carrier protein. Linker proteolysis can be excluded as a cause for the two bands since biotinylation of the Avi-tag with biotin ligase similarly generated two distinct bands (Supplementary Data and Figure S2).