Targeting a heterologous protein to multiple plant organelles via rationally designed 5′ mRNA tags

Journal of Biological Engineering

Matthew D Mattozzi²  Jeffrey C Way²  Pamela A Silver²  Mathias J Voges¹ 

[1] Department of Biotechnology, Technische Universiteit Delft, 2628 BC, Delft, Netherlands;Chimerion Biotechnology Inc, Cambridge, MA 02138, USA

关键词: Nicotiana benthamiana;    Organelle;    Cytosol;    Chloroplast;    Peroxisome;    Localization;    Signal embedding;    Alternative splicing;    Protein targeting;   

Others  :  805320
DOI  :  10.1186/1754-1611-7-20

 received in 2013-03-06, accepted in 2013-08-17,  发布年份 2013

【 摘 要 】

Background

Plant bioengineers require simple genetic devices for predictable localization of heterologous proteins to multiple subcellular compartments.

Results

We designed novel hybrid signal sequences for multiple-compartment localization and characterize their function when fused to GFP in Nicotiana benthamiana leaf tissue. TriTag-1 and TriTag-2 use alternative splicing to generate differentially localized GFP isoforms, localizing it to the chloroplasts, peroxisomes and cytosol. TriTag-1 shows a bias for targeting the chloroplast envelope while TriTag-2 preferentially targets the peroxisomes. TriTag-3 embeds a conserved peroxisomal targeting signal within a chloroplast transit peptide, directing GFP to the chloroplasts and peroxisomes.

Conclusions

Our novel signal sequences can reduce the number of cloning steps and the amount of genetic material required to target a heterologous protein to multiple locations in plant cells. This work harnesses alternative splicing and signal embedding for engineering plants to express multi-functional proteins from single genetic constructs.

【 授权许可】

   
2013 Voges et al.; licensee BioMed Central Ltd.

【 图 表 】

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