(A) SDS-PAGE gel of proteins expressed from Expression Clones of the genes in pDEST17 (His6 fusion) vector in Escherichia coli strain BL21 SI. (Lane 1), eIF4e; (lane 2), tyrosine kinase; (lane 3), transferrin receptor; (lane4), β-adaptin; (lane 5), MAP4 (MAP4 is known to migrate aberrantly on SDS-PAGE gels; Chapin et al. 1995); (lane6), Gus. (B) SDS-PAGE gel of expression of genes subcloned from the same Entry Clones as in (A) into baculovirus Destination Vectors. (Lanes 1–5) are pDest8 (native expression; (lane 1), eIF4e; (lane2), tyrosine kinase; (lane 3), transferrin receptor; (lane 4), β-adaptin; (lane 5), MAP4). (Lanes 6–8) the gus gene, from the same Entry Clone as in A, expressed in the baculovirus vectors pDest8 (lane 6, native, 68 kD), pDest10 (lane 7, His6 fusion, 74 kD), and pDest20 (lane 8, GST fusion, 97 kD). As inE. coli expression (above), MAP4 protein migrated as a ∼200-kD protein instead of as a 121-kD protein (Chapin et al. 1995).
