Combinatorial codon scrambling enables scalable gene synthesis and amplification of repetitive proteins.
Journal Article (Journal Article)
Most genes are synthesized using seamless assembly methods that rely on the polymerase chain reaction (PCR). However, PCR of genes encoding repetitive proteins either fails or generates nonspecific products. Motivated by the need to efficiently generate new protein polymers through high-throughput gene synthesis, here we report a codon-scrambling algorithm that enables the PCR-based gene synthesis of repetitive proteins by exploiting the codon redundancy of amino acids and finding the least-repetitive synonymous gene sequence. We also show that the codon-scrambling problem is analogous to the well-known travelling salesman problem, and obtain an exact solution to it by using De Bruijn graphs and a modern mixed integer linear programme solver. As experimental proof of the utility of this approach, we use it to optimize the synthetic genes for 19 repetitive proteins, and show that the gene fragments are amenable to PCR-based gene assembly and recombinant expression.
Full Text
Duke Authors
Cited Authors
- Tang, NC; Chilkoti, A
Published Date
- April 2016
Published In
Volume / Issue
- 15 / 4
Start / End Page
- 419 - 424
PubMed ID
- 26726995
Pubmed Central ID
- PMC4809025
Electronic International Standard Serial Number (EISSN)
- 1476-4660
International Standard Serial Number (ISSN)
- 1476-1122
Digital Object Identifier (DOI)
- 10.1038/nmat4521
Language
- eng