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CBI has applied sophisticated technologies to construct completely synthetic “designer genes” expressing the exact desired protein. Genes are synthesized by combining overlapping complementary oligonucleotides, filling in the gaps with a thermostable nucleotide polymerase, and amplifying the product by PCR. These amplified products are cloned into the desired plasmid vector and overexpressed. Genes up to ~2.0 kbp have been synthesized, but there is no theoretical limit to the size. Synthetic “designer genes” normally have unique DNA sequences that have been modified to optimize the codon usage for the target host. In addition, restriction sites, leader sequences, homing signals, or other modified sequences can be incorporated for special applications.
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