ori
 A cm

 Ap
 bA7
 b+7

 pTAM147 )V5( pTAM35

 ori3.9 kb 6 .9 kb




 bA7 -V5 b+7-h is





 B bwt-his
 bwt-vs

 N- b
 +7-his
 terminus
 bw-v
 b+7-his
 b
 A7-V5

Figure 2-8. Histidine and V5-epitope-tagged b subunit expression system. A) Both ba7~-vs
 and b+7-his were expressed together in KM2 (Ab) cells using a two-plasmid
 expression system. Plasmid pTAM47 was designed to express high levels of
 V5-tagged ba7. This plasmid contains the genes conferring ampicillin
 resistance and the pUC18 origin of replication. Plasmid pTAM35 includes the
 chloramphenicol resistance gene, the pACYC 184 origin of replication and
 expresses histidine-tagged b+7. In similar constructions, plasmids pTAM37
 (bwt-his, Cmr) and pTAM46 (bwt4.xs, Apr), or pTAM35 (b+7-his, Cmr) and
 pTAM46 (bwt4.xs, Apr) were developed for coexpression experiments. B) Line
 diagram of the different b subunits that were coexpressed in the same cell.
 Interactions between birt-his / b,,.-VS, b+7-his / b,,.-VS, and b+7-his /bA7~-V5 were
 investigated.