W0205

Whole Gene Synthesis and High Throughput Crystallography. Alex B. Burgin, John Walchli, Kathryn Hjerrild, and Lance Stewart, BioStructures Group, deCODE Genetics, 7869 NE Day Rd W, Bainbridge Island, WA 98110.

Many different bottlenecks must be solved before gene to structure high throughput crystallography can be an efficient way to solve new protein structures. One primary bottleneck is rapidly and efficiently creating gene constructs that are likely to express proteins at high levels and are likely to provide proteins that can be crystallized. In order to overcome this bottleneck, we have developed software and associated technology for the design and synthesis of genes from synthetic oligonucleotides. The process is cost effective and standardizes the gene construction step thereby dramatically increasing the number of gene constructs that can be generated in a short period of time. The resulting genes are codon optimized for a given expression system thereby increasing the likelihood of obtaining protein in high yield. The process also allows the precise engineering of protein constructs without relying upon existing restriction sites or obtaining a cDNA clone.