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A dynamic, genome-scale flux model of Lactococcus lactis to increase specific recombinant protein expression

Paper ID Volume ID Publish Year Pages File Format Full-Text
31797 44839 2009 15 PDF Available
Title
A dynamic, genome-scale flux model of Lactococcus lactis to increase specific recombinant protein expression
Abstract

Recently, lactic acid bacteria (LAB) have attracted a great deal of interest because of their potential to serve as oral delivery vehicles for recombinant protein vaccines. An important limitation to their use is the typically low level of heterologous expression obtained in LAB. To address this, a dynamic flux balance analysis (DFBA) model was used to identify gene targets for increasing specific expression of Green Fluorescent Protein (GFP), a model heterologous protein, in Lactococcus lactis IL1403. Two strains, each targeting one of the top model-identified genes, were constructed and tested in vivo. Data show that both strains, by a conservative estimate, achieved 15% higher GFP per cell than the control strain, a qualitative confirmation of the model predictions. A genome-scale DFBA model for L. lactis growing on M17 medium is presented along with the procedure for screening gene targets and a powerful method for visualizing fluxes in genome-scale metabolic networks.

Keywords
Lactococcus lactis; GFP; pMSP3535H3; Flux balance; DFBA; Metabolic engineering
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A dynamic, genome-scale flux model of Lactococcus lactis to increase specific recombinant protein expression
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 11, Issue 6, November 2009, Pages 367–381
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us