Production and secretion of a heterologous protein by turnip hairy roots with superiority over tobacco hairy roots

Abstract

A fully contained and efficient heterologous protein production system was designed using Brassica rapa rapa (turnip) hairy roots. Two expression cassettes containing a cauliflower mosaic virus (CaMV) 35S promoter with a duplicated enhancer region, an Arabidopsis thaliana sequence encoding a signal peptide and the CaMV polyadenylation signal were constructed. One cassette was used to express the green fluorescent protein (GFP)-encoding gene in hairy roots grown in flasks. A stable and fast-growing hairy root line secreted GFP at >120 mg/l culture medium. GFP represented 60 % of the total soluble proteins in the culture medium. Turnip hairy roots retained sustainable growth and stable GFP production over 3 years. These results were superior to those obtained using tobacco hairy roots.

Production of glucosinolates, phenolic compounds and associated gene expression profiles of hairy root cultures in turnip (Brassica rapa ssp. rapa)

“…Previously it was reported that rolC from A. rhizogenes T-DNA could stimulate the production of secondary metabolites in the transformed cells of different plants (Shkryl et al 2008). Earlier limited studies have been reported in hairy root induction of turnip (Tanaka et al 1985; Huet et al 2014). However, only a few studies have been described in the individual GSLs content of hairy root cultures in Nasturtium officinale (Park et al 2011), Sinapis alba and B. rapa (Kastell et al 2013b), B. oleracea var.…”

In vitro regeneration and Agrobacterium mediated genetic transformation of Artemisia aucheri Boiss

“…Genetically transformed roots (so called hairy roots) mediated with Agrobacterium rhizogenes creates a rapid and simple tool to integrate and express foreign genes in plant cells (Georgiev et al 2007;Sharafi et al 2013aSharafi et al , 2014aHuet et al 2014). The molecular basis of hairy root induction is a transfer DNA (T-DNA) from root inducing (Ri) plasmid of A. rhizogenes.…”

Determination of Multimodal Isotopic Distributions: The Case of a15N Labeled Protein Produced into Hairy Roots

“…15 eGFP Production. Hairy root selection, culture media preparation, and eGFP production were achieved with a protocol similar to the one published by Huet et al, 8 as described in the Supporting Information (Experimental Methods).…”

“…Furthermore, they are able to secrete the protein of interest in high amount (about 120 mg L −1 ) in the culture media thanks to the fusion of a signal peptide, making protein purification easier to achieve. 8 In this context, we have recently initiated a program aiming at exploring the possibility to produce, in hairy roots, labeled glycoproteins for NMR structural studies. One milestone of our project consisted of checking the abilities of roots to produce 15 N or 15 N/ 13 C labeled proteins, and our first investigations focused on the 15 N labeling.…”