Gateway-Assisted Vector Construction to Facilitate Expression of Foreign Proteins in the Chloroplast of Single Celled Algae

Melanie Oey,Ian L Ross,Ben Hankamer

doi:10.1371/journal.pone.0086841

Melanie Oey, Ian L Ross + Show 1 more

Open Access

https://doi.org/10.1371/journal.pone.0086841

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Journal: PLoS ONE	Publication Date: Feb 11, 2014
Citations: 36	License type: CC BY 4.0

Affiliation: University of Queensland

Abstract

With a rising world population, demand will increase for food, energy and high value products. Renewable production systems, including photosynthetic microalgal biotechnologies, can produce biomass for foods, fuels and chemical feedstocks and in parallel allow the production of high value protein products, including recombinant proteins. Such high value recombinant proteins offer important economic benefits during startup of industrial scale algal biomass and biofuel production systems, but the limited markets for individual recombinant proteins will require a high throughput pipeline for cloning and expression in microalgae, which is currently lacking, since genetic engineering of microalgae is currently complex and laborious. We have introduced the recombination based Gateway® system into the construction process of chloroplast transformation vectors for microalgae. This simplifies the vector construction and allows easy, fast and flexible vector design for the high efficiency protein production in microalgae, a key step in developing such expression pipelines.

Highlights

By 2050, world population is projected to reach 9.6 billion and with continued economic development the demand for food and energy is forecast to increase by 70% (UN) and 50% (International Energy Agency), respectively
Using green fluorescent protein (GFP) as a demonstration system, we have utilised the GatewayH system to create a simple, flexible, easy to modify and interchangeable vector system that allows the rapid insertion of different expression cassettes into the destination vectors of choice
To harness microalgae for the efficient foreign protein production, chloroplast transformation is the method of choice, providing high expression levels

Summary

Introduction

By 2050, world population is projected to reach 9.6 billion and with continued economic development the demand for food and energy is forecast to increase by 70% (UN) and 50% (International Energy Agency), respectively. GatewayH site-specific recombination system Traditional cloning of each new gene into a suitable antibiotic resistance vector is an obstacle to rapid strain development. Using green fluorescent protein (GFP) as a demonstration system, we have utilised the GatewayH system to create a simple, flexible, easy to modify and interchangeable vector system that allows the rapid insertion of different expression cassettes into the destination vectors of choice.

Results

Conclusion