DEVELOPMENT OF A SYSTEM FOR HIGH-SPEED GENE SYNTHESIS

Egea Biosciences, Inc., San Marcos CA

Investigators

Abstract

The Human Genome Sequencing Project is generating a vast amount of raw sequence information. However, a limitation in the analysis of new human proteins is the difficulty of converting digital sequence information into "wet" material in the laboratory. Conventional recombinant cloning is expensive, time consuming and limited in scope. We propose utilizing large scale oligonucleotide synthesis to address this need and will develop a high-throughput gene synthesizer, an instrument capable of converting a DNA or protein sequence designed in the computer into long DNA molecules for protein expression. A prototype system consists of: 1) designing, editing, databasing, and modifying gene and vector sequences in silico; 2) generating a set of overlapping oligonucleotide sequences covering the entire design; 3) programming automated synthesis of the oligonucleotide set; 4) robotically assembling the component oligonucleotides and ligating these into large DNA strands; and 5) transforming the resulting DNA into E. coli for gene replication and protein expression. The prototype system is capable of producing plasmids expressing recombinant human proteins at one gene/instrument/day and acceptable cost. Using this approach it is conceivable to produce most or all human proteins as arrays of recombinant bacteria for further functional analysis. PROPOSED COMMERCIAL APPLICATIONS: This work will result in 1) a prototype instrument for gene and genome synthesis that could be the basis of a commercial product; and 2) a technology for high throughput gene and protein production that could result in commercial applications of human protein arrays for functional and structural analysis.