Complementation of the yeast rer2Δ mutant
To express CPT3 and LEW1 in Saccharomyces
cerevisiae mutant cells (rer2 Δ mutant: rer2 ::kanMX4ade2-101 ura3-52 his3-200 lys2-801 ), coding
sequences of CPT3 and LEW1 (AT1G11755) were subcloned into
the pESC-URA yeast dual expression vector (Agilent, Santa Clara, CA,
USA) according to the manufacturer’s protocol. Transformant selection
and growth, as well as analyses of polyisoprenoid profile and CPY
glycosylation status, were performed as described previously
(Surowiecki, Onysk, Manko, Swiezewska, & Surmacz, 2019).