1469-896X/asset/olbannerleft.gif?v=1&s=d218899ae53b2862ab119790ed504b8d72122fb3)
1469-896X/asset/olbannerright.gif?v=1&s=59470eb9a1d9b7b13b1be75e9445e6c46ee2214f)
research-article
Protein fabrication automation
Article first published online: 2 JAN 2009
DOI: 10.1110/ps.062591607
Copyright © 2007 The Protein Society
1469-896X/asset/cover.gif?v=1&s=0ff483325083001100314f63c633a394aff24478 "Protein Science")
Additional Information
How to Cite
Cox, J. C., Lape, J., Sayed, M. A. and Hellinga, H. W. (2007), Protein fabrication automation. Protein Science, 16: 379–390. doi: 10.1110/ps.062591607
Publication History
- Issue published online: 2 JAN 2009
- Article first published online: 2 JAN 2009
- Manuscript Revised: 25 NOV 2006
- Manuscript Accepted: 25 NOV 2006
- Manuscript Received: 28 SEP 2006
Keywords:
- gene assembly;
- automation;
- synthetic ORF;
- protein expression;
- fabrication
Abstract
Facile “writing” of DNA fragments that encode entire gene sequences potentially has widespread applications in biological analysis and engineering. Rapid writing of open reading frames (ORFs) for expressed proteins could transform protein engineering and production for protein design, synthetic biology, and structural analysis. Here we present a process, protein fabrication automation (PFA), which facilitates the rapid de novo construction of any desired ORF from oligonucleotides with low effort, high speed, and little human interaction. PFA comprises software for sequence design, data management, and the generation of instruction sets for liquid-handling robotics, a liquid-handling robot, a robust PCR scheme for gene assembly from synthetic oligonucleotides, and a genetic selection system to enrich correctly assembled full-length synthetic ORFs. The process is robust and scalable.