Gene Synthesis
The 897-bp portion of the DNA sequence of BTE lacking the 83 amino acids at the Nterminus
that appear to be involved in thylakoid targeting (Voelker et al., 1992) and
containing the XbaI site formerly used to clone a functional part of the gene in E. coli
(Ohlrogge et al., 1995) was codon-optimized for expression in E. coli, with common
restriction sites eliminated. An artificial ribosome-binding site (RBS; AGGAGG), spacer
sequence, start codon, and bases to create an in-frame sequence were added upstream of the
gene fragment. The full sequence (Supplementary Material) was custom-synthesized
(Integrated DNA Technologies, Inc.) and was received in plasmid pUC57 (pUC57-BTE).