Protein expression from synthetic genes: selection of clones using GFP.

Protein expression from synthetic genes: selection of clones using GFP. Research Abstract Details 

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Protein expression from synthetic genes: selection of clones using GFP. Abstract Text:

Sandrine Denis-Quanquin,Lucille Lamouroux, Lougarre,Sabrina ,Isabelle Saves,Laurent Paquereau,Pascal Demange,Didier Fournier,

Construction of synthetic genes is today the most elegant way to optimize the heterologous expression of a recombinant protein. However, the selection of positive clones that incorporate the correct synthetic DNA fragments is a bottleneck as current methods of gene synthesis introduce 3.5 nucleotide deletions per kb. Furthermore, even when all predictable optimizations for protein production have been introduced into the synthetic gene, production of the protein is often disappointing: protein is produced in too low amounts or end up in inclusion bodies. We propose a strategy to overcome these two problems simultaneously by cloning the synthetic gene upstream of a reporter gene. This permits the selection of clones devoid of frame-shift mutations. In addition, beside nucleotide deletion, an average of three non-neutral mutations per kb are introduced during gene synthesis. Using a reporter protein downstream of the synthetic gene, allows the selection of clones with random mutations improving the expression or the folding of the protein of interest. The problem of errors found in synthetic genes is then turned into an advantage since it provides polymorphism useful for molecular evolution. The use of synthetic genes appears as an alternative to the error-prone PCR strategy to generate the variations necessary in protein engineering experiments.

Protein expression from synthetic genes: selection of clones using GFP. Publishing Authors By Initials

S Denis-Quanquin,L Lamouroux,A Lougarre,S ,I Saves,L Paquereau,P Demange,D Fournier,

For similar proteins: recombinant proteins: recombinant fusion proteins research abstracts see: proteins: recombinant proteins: recombinant fusion proteins research

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Protein expression from synthetic genes: selection of clones using GFP. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of biotechnology

VOLUME: 131

Page Numbers: 223-30

Journal Abbreviation: J. Biotechnol.

ISSN: 0168-1656

DAY: 21

MONTH: 07

YEAR: 2007

Protein expression from synthetic genes: selection of clones using GFP. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 8411927

Protein expression from synthetic genes: selection of clones using GFP. Keywords Mesh Terms:

KEYWORDS: Recombinant Fusion Proteins

MESH TERMS: metabolism

Chemical & Substance for Abstract: Protein expression from synthetic genes: selection of clones using GFP. Information

Substance Name: Green Fluorescent Proteins

Registry Number: 147336-22-9

Grant and Affiliation Information for Protein expression from synthetic genes: selection of clones using GFP.

AFFILIATION: IPBS, 205 route de Narbonne, 31000 Toulouse, France.

Country: Netherlands

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MEDLINETA: J Biotechnol

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