Figure 2.

Functional characterization of rapeseed LPAATs. A, Complementation of an acyltransferase-deficient strain of E. coli. The coding sequences corresponding to the BAT1.13 and BAT1.5 cDNAs were cloned into the pBluescript SK+ vector and transformed into the JC201 plsC⁻ strain. Aliquots from OD₆₀₀ = 0.5 cultures were grown in the presence of IPTG. The figure shows the OD after 16 h of growth at various temperatures. JC201 indicates strain only; pBSK indicates strain JC201 transformed with vector only; BAT1.13 and BAT1.5 indicate strain JC201 transformed with the vector containing the coding sequence of BAT1.13 or BAT1.5. Data represent means of three independent transformations. B, Rapeseed microsomal LPAAT activities in E. coli membrane preparations. Membranes were isolated from JC201 cells that had been grown at 30°C after induction by IPTG and were assayed in the presence of saturating concentrations of LPA and [1-¹⁴C]oleoyl-CoA or [1-¹⁴C]palmitoyl-CoA, and the quantity of radioactivity in the PA reaction product was integrated. The genotypes of JC201, pBSK, BAT1.13, and BAT1.5 are as in A. The values represent means of three membrane preparations from independent transformant clones.