Protein fabrication automation

被引：44

作者：

Cox, J. Colin ^{[1
]}

Lape, Janel ^{[1
]}

Sayed, Mahmood A. ^{[1
]}

Hellinga, Homme W. ^{[1
]}

机构：

[1] Duke Univ, Med Ctr, Dept Biochem, Durham, NC 27710 USA

来源：

PROTEIN SCIENCE | 2007年 / 16卷 / 03期

关键词：

gene assembly; automation; synthetic ORF; protein expression; fabrication; ESCHERICHIA-COLI; GENE SYNTHESIS; IN-VIVO; COMPUTATIONAL DESIGN; SYNTHETIC GENES; DNA-SYNTHESIS; HIGH-FIDELITY; CONSTRUCTION; EXPRESSION; SEQUENCES;

D O I：

10.1110/ps.062591607

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：379 / 390

页数：12

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