Figure 4.

Expression and activity of LeFPS1 in recombinant E. coli. A, SDS-PAGE analysis of 50 μg of total protein extracts from bacteria transformed with the non-recombinant pET28a vector or with the pET-LeFPS plasmid with (+) or without (−) IPTG induction. After Coomassie Blue staining, a new polypeptide (approximately 40 kD) was detected in IPTG-induced bacteria containing the pET-LeFPS plasmid. B, Sonicated bacterial extracts were incubated with [¹⁴C]IPP and GPP. Isoprenoids were extracted, dephosphorylated, and separated on thin layer chromatography prior to autoradiography. Radiolabeled products were identified after GLC analysis. Lanes are as indicated in A. C, Quantification of [¹⁴C]farnesol produced by recombinant bacteria protein extracts. After thin layer chromatographic analysis and autoradiography, farnesol spots were collected and radioactivity was estimated. Results are the average of three independent experiments and are expressed in picomoles of FPP produced per microgram of total bacterial protein. Bars indicate the sd. Lanes are as indicated in A.