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. 2014 Jan 23;9(1):e86482. doi: 10.1371/journal.pone.0086482

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© 2014 Hung et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

PMC Copyright notice

The GB1-NS4A(1–48) sequence was inserted into pGEX-4T-2 between the EcoRI and XhoI sites. Protease recognition motifs are underlined while the cleavage sites are marked by triangles. The two thrombin sites, which originate from the vector backbone of pGEX and pGEV, respectively, are shown in brackets and were not used in our protocol. Note that TEV digestion produces a native NS4A peptide.