Figure 5

Overview of cloning and expression results. All cloning results with the described vectors and targets are summarized. Two colonies were picked for each vector-target combination and analyzed in order to apply a stringent test to the cloning and expression systems. Top lanes state vector name and design features; vertical lanes indicate target proteins; shaded lanes indicate experimental results; code see below; upper lane with cloning result, ie. PCR with insert primers; lower lane with expression result, ie. soluble protein after mini-IMAC target purification. Positive Test PCR means a clearly visible band of the expected size on agarose gel. Positive for soluble protein means a clearly visible band on the Coomassie-stained SDS-PAGE at the correct size in the fraction after purification by Ni-NTA column chromatography. Designations: 1/2 and 2/2 indicate first and second clone that were picked; CapGly, Mr = 10 KDa, CAP-Gly domain 1 of human CLIP170, accession number NP_002947; EB1, full length of human EB1 protein, Mr = 32 KDa, accession number AAC09471; TTL, Mr = 48 KDa, full length human tubulin-tyrosin ligase, accession number NP_714923; VP3, Mr = 62 KDa, full length of adeno-associated virus capsid protein 3, accession number AF043303, synthetic sequence, see 'Additional file 1'; PKNG, 78 KDa, full length of the M. tuberculosis serine/threonine-protein kinase G, accession number NP_214924; CLIP170, Mr = 54 KDa, fragment of human CLIP170 fused with a GCN4 sequence, accession number NP_002947.