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. Author manuscript; available in PMC: 2013 Apr 1.
Published in final edited form as: Protein Expr Purif. 2012 Feb 8;82(2):317–324. doi: 10.1016/j.pep.2012.01.019

Figure 2. Protein sequence alignment of the CBS constructs.

Figure 2

Bold sequence marked by asterisks designates CBS sequence that is conserved among the constructs. The pGEX-6P1-hCBS yields fusion protein with the GST (semibold italic) and the HRV3C protease recognition site (italic) at the N-terminus of CBS. The pET28-C-hCBS and pET28-N-hCBS constructs carry permanent 6xHis tag (underlined) at the C-terminus and the N-terminus of CBS, respectively. The pET47-NP-hCBS construct yields CBS enzyme with 6xHis tag and HRV3C protease recognition site at the N-terminus of CBS. The GST partner or 6xHis tag can be removed after cleavage with HRV3C protease (designated by an arrow) yielding native CBS sequence with only one extra amino acid (glycine) at the N-terminus from the pGEX-6P1-hCBS and the pET47-NP-hCBS constructs. Dashes represent gaps in protein alignment, while dots stand for the remaining sequence of either GST or CBS.