Protein body formation in stable transgenic tobacco expressing elastin-like polypeptide and hydrophobin fusion proteins

BMC Biotechnology

Sonia P Gutiérrez¹  Reza Saberianfar¹  Susanne E Kohalmi²  Rima Menassa¹ 

[1] Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada, London, ON, Canada

[2] Department of Biology, University of Western Ontario, London, ON, Canada

关键词: Green fluorescent protein;    Molecular farming;    Transgenic expression;    Tobacco;    HFBI;    Hydrophobin I;    ELP;    Elastin-like polypeptide;    Protein body formation;    Protein body;   

Others  :  1123174
DOI  :  10.1186/1472-6750-13-40

 received in 2013-02-06, accepted in 2013-05-06,  发布年份 2013

【 摘 要 】

Background

Plants are recognized as an efficient and inexpensive system to produce valuable recombinant proteins. Two different strategies have been commonly used for the expression of recombinant proteins in plants: transient expression mediated by Agrobacterium; or stable transformation of the plant genome. However, the use of plants as bioreactors still faces two main limitations: low accumulation levels of some recombinant proteins and lack of efficient purification methods. Elastin-like polypeptide (ELP), hydrophobin I (HFBI) and Zera® are three fusion partners found to increase the accumulation levels of recombinant proteins and induce the formation of protein bodies (PBs) in leaves when targeted to the endoplasmic reticulum (ER) in transient expression assays. In this study the effects of ELP and HFBI fusion tags on recombinant protein accumulation levels and PB formation was examined in stable transgenic Nicotiana tabacum.

Results

The accumulation of recombinant protein and PB formation was evaluated in two cultivars of Nicotiana tabacum transformed with green fluorescent protein (GFP) fused to ELP or HFBI, both targeted and retrieved to the ER. The ELP and HFBI tags increased the accumulation of the recombinant protein and induced the formation of PBs in leaves of stable transgenic plants from both cultivars. Furthermore, these tags induced the formation of PBs in a concentration-dependent manner, where a specific level of recombinant protein accumulation was required for PBs to appear. Moreover, agro-infiltration of plants accumulating low levels of recombinant protein with p19, a suppressor of post-transcriptional gene silencing (PTGS), increased accumulation levels in four independent transgenic lines, suggesting that PTGS might have caused the low accumulation levels in these plants.

Conclusion

The use of ELP and HFBI tags as fusion partners in stable transgenic plants of tobacco is feasible and promising. In a constitutive environment, these tags increase the accumulation levels of the recombinant protein and induce the formation of PBs regardless of the cultivar used. However, a specific level of recombinant protein accumulation needs to be reached for PBs to form.

【 授权许可】

   
2013 Gutiérrez et al.; licensee BioMed Central Ltd.

【 图 表 】

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