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. Author manuscript; available in PMC: 2017 Aug 4.
Published in final edited form as: Methods Enzymol. 2015 Sep 6;569:63–78. doi: 10.1016/bs.mie.2015.08.011

Figure 1.

Figure 1

Schematic drawing of the expression constructs for human SUN2 (A), and KASH (B). Each SUN2 fragment includes the SUN domain (residues 540-717) and preceding lumenal segments of different length, predicted to form coiled-coils (CC). The SUN2 (residue 335-717) fragment is N-terminally fused to a 3C-cleavable 6xHis tag, whereas a 6xHis:triGCN4 tag is used for a shorter SUN2 fragment (residues 522-717). KASH motifs are C-terminally attached to 3C-cleavable MBP, superfolder GFP (sfGFP) or GB1 tags. Short flexible linkers are incorporated around the 3C cleavage sites to enhance removal of the fusion tags.