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. 2004 Sep;87(3):1795–1804. doi: 10.1529/biophysj.104.039842

FIGURE 1.

FIGURE 1

(A) Amino acid sequence alignment for mouse KIF3A and KIF3B and human conventional kinesin heavy chain (HsKHC) genes at the neck-coil junction. Secondary structure predictions were taken from the rat KHC crystal structure (Kozielski et al., 1997) and the start of the coiled-coil of KIF3A and KIF3B was inferred by comparison to the conventional kinesin sequence and by predictions from the COILS program. There is an obvious splice site in the neck-linker region of KIF3A and KIF3B; the arrow denotes where the AflII restriction site was introduced. (B) Constructing mutant KIF3A/A. KIF3A and KIF3B plasmids were digested and the sequence for the KIF3A head domain was spliced to the sequence for the KIF3B rod and tail domains. This chimaeric gene was then coexpressed with the wild-type KIF3A gene in insect cells, producing a mutant protein that has two KIF3A heads and the normal KIF3A/B rod and tail structure. An analogous approach was used to make KIF3B/B. GenBank accession numbers: KIF3A, NM_008443; KIF3B, NM_008444; HsKHC, X65873.