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. 2013 Jul 27;1091:131–141. doi: 10.1007/978-1-62703-691-7_8

Fig. 3.

Fig. 3

Multiprotein complexes produced from polyproteins. (a) The TAF8/TAF10 dimer (inserted into the Tn7 attachment site) was co-expressed as a polyprotein with the yellow fluorescent protein (YFP) inserted into the viral LoxP site from a composite baculovirus (EMBacY-TAF8/TAF10). YFP and CFP expression per one million cells were tracked for evaluating the viral infection and polyprotein production. St stands for a defined fluorescence standard (used to calibrate for 100,000 arbitrary units), dpa stands for day of proliferation arrest in the infected culture [5]. (b) SDS-PAGE (left ) shows balanced expressions of TAF8 and TAF10. Complete proteolysis of the TAF8/TAF10 polyprotein was confirmed by Western blot (right ) using antibody specific for the hexa histidine-tags of TAF10 and TEV protease (doublet ). M stands for molecular weight marker, C stands for cell control (uninfected insect cells), W stands for whole cell extract, S stands for supernatant. (c) Sections from SDS-PAGE are shown for TAF8/TAF10 dimer from size exclusion chromatography purification, SMAT complex from IMAC batch purification [5], 3TAF and core-TFIID complexes from size exclusion chromatography purification [4]