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A highly parallel method for synthesizing DNA repeats enables the discovery of 'smart' protein polymers.

Publication ,  Journal Article
Amiram, M; Quiroz, FG; Callahan, DJ; Chilkoti, A
Published in: Nat Mater
February 2011

Robust high-throughput synthesis methods are needed to expand the repertoire of repetitive protein-polymers for different applications. To address this need, we developed a new method, overlap extension rolling circle amplification (OERCA), for the highly parallel synthesis of genes encoding repetitive protein-polymers. OERCA involves a single PCR-type reaction for the rolling circle amplification of a circular DNA template and simultaneous overlap extension by thermal cycling. We characterized the variables that control OERCA and demonstrated its superiority over existing methods, its robustness, high-throughput and versatility by synthesizing variants of elastin-like polypeptides (ELPs) and protease-responsive polymers of glucagon-like peptide-1 analogues. Despite the GC-rich, highly repetitive sequences of ELPs, we synthesized remarkably large genes without recursive ligation. OERCA also enabled us to discover 'smart' biopolymers that exhibit fully reversible thermally responsive behaviour. This powerful strategy generates libraries of repetitive genes over a wide and tunable range of molecular weights in a 'one-pot' parallel format.

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Published In

Nat Mater

DOI

EISSN

1476-4660

Publication Date

February 2011

Volume

10

Issue

2

Start / End Page

141 / 148

Location

England

Related Subject Headings

  • Proteins
  • Protein Engineering
  • Nanoscience & Nanotechnology
  • Glucagon-Like Peptide 1
  • Genes, Synthetic
  • Gene Library
  • Elastin
  • DNA, Circular
  • DNA
  • Biopolymers
 

Citation

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Amiram, M., Quiroz, F. G., Callahan, D. J., & Chilkoti, A. (2011). A highly parallel method for synthesizing DNA repeats enables the discovery of 'smart' protein polymers. Nat Mater, 10(2), 141–148. https://doi.org/10.1038/nmat2942
Amiram, Miriam, Felipe Garcia Quiroz, Daniel J. Callahan, and Ashutosh Chilkoti. “A highly parallel method for synthesizing DNA repeats enables the discovery of 'smart' protein polymers.Nat Mater 10, no. 2 (February 2011): 141–48. https://doi.org/10.1038/nmat2942.
Amiram M, Quiroz FG, Callahan DJ, Chilkoti A. A highly parallel method for synthesizing DNA repeats enables the discovery of 'smart' protein polymers. Nat Mater. 2011 Feb;10(2):141–8.
Amiram, Miriam, et al. “A highly parallel method for synthesizing DNA repeats enables the discovery of 'smart' protein polymers.Nat Mater, vol. 10, no. 2, Feb. 2011, pp. 141–48. Pubmed, doi:10.1038/nmat2942.
Amiram M, Quiroz FG, Callahan DJ, Chilkoti A. A highly parallel method for synthesizing DNA repeats enables the discovery of 'smart' protein polymers. Nat Mater. 2011 Feb;10(2):141–148.

Published In

Nat Mater

DOI

EISSN

1476-4660

Publication Date

February 2011

Volume

10

Issue

2

Start / End Page

141 / 148

Location

England

Related Subject Headings

  • Proteins
  • Protein Engineering
  • Nanoscience & Nanotechnology
  • Glucagon-Like Peptide 1
  • Genes, Synthetic
  • Gene Library
  • Elastin
  • DNA, Circular
  • DNA
  • Biopolymers