Journal Article

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

Gutiérrez, Sonia P, Menassa, Rima, Saberianfar, Reza, Kohalmi, Susanne E

2013

Overview

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.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V
• Subject: Accumulation / Agrobacterium / agroinfiltration / Amino acids / Applied Microbiology / Biochemical Engineering / Biomedical Engineering/Biotechnology / Bioreactors / Biotechnology / Chemistry / Chemistry and Materials Science / Cultivars / Elastin / Elastin - biosynthesis / Elastin - genetics / Elastin - metabolism / endoplasmic reticulum / Endoplasmic Reticulum - chemistry / Endoplasmic Reticulum - metabolism / Farm buildings / Fluorescence / Food / Fungal Proteins - biosynthesis / Fungal Proteins - genetics / Fungal Proteins - metabolism / Fungi / genes / Genetic Engineering / genetically modified organisms / Genetically modified plants / green fluorescent protein / Green Fluorescent Proteins - analysis / Green Fluorescent Proteins - biosynthesis / Green Fluorescent Proteins - genetics / Green Fluorescent Proteins - metabolism / Hydrophobins / Leaves / Nicotiana - genetics / Nicotiana - metabolism / Nicotiana tabacum / Phase transitions / Physiological aspects / Plant biotechnology / Plant Breeding/Biotechnology / Plant Leaves - chemistry / Plant Leaves - metabolism / Plants, Genetically Modified - genetics / Plants, Genetically Modified - metabolism / Polypeptides / protein bodies / Protein Engineering / Proteins / Recombinant Fusion Proteins - analysis / Recombinant Fusion Proteins - biosynthesis / Recombinant Fusion Proteins - genetics / Recombinant Fusion Proteins - metabolism / Recombinant proteins / Research Article / RNA Interference / Statistical analysis / tobacco / Transgenic plants