COPE
Volume 4, Number 3 (Jul 2016)                   Res Mol Med (RMM) 2016, 4(3): 35-40 | Back to browse issues page



DOI: 10.18869/acadpub.rmm.4.3.35

XML Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Zangi M, Ofoghi H, Amini-Bayat Z, Ehsani P. Utility of P19 Gene-Silencing Suppressor for High Level Expression of Recombinant Human Therapeutic Proteins in Plant Cells. Res Mol Med (RMM). 2016; 4 (3) :35-40
URL: http://rmm.mazums.ac.ir/article-1-206-en.html

Abstract:   (1438 Views)

Background: The potential of plants, as a safe and eukaryotic system, is considered in the production of recombinant therapeutic human protein today; but the expression level of heterologous proteins is limited by the post-transcriptional gene silencing (PTGS) response in this new technology. The use of viral suppressors of gene silencing can prevent PTGS and improve transient expression level of foreign proteins. In this study, we investigated the effect of p19 silencing suppressor on recombinant human nerve growth factor expression in Nicotiana benthamiana.

Materials and Methods: The p19 coding region was inserted in the pCAMBIA using NcoI and BstEII recognition sites. Also, the cloned synthesized recombinant human NGF (rhNGF) fragment was cloned directly into PVX vector by ClaI and SalI restriction enzymes. The co-agroinfiltration of rhNGF with p19 viral suppressor of gene silencing was evaluated by dot-blot and SDS-PAGE. The amount of expressed rhNGF protein was calculated by AlphaEaseFC software.

Results: Co-agroinfiltration of hNGF with P19 suppressor showed about forty-fold increase (8% total soluble protein (TSP)) when compared to the absence of P19 suppressor (0.2%TSP).

Conclusion: The results presented here confirmed that the use of P19 gene silencing suppressor derived from tomato bushy stunt virus (TBSV) could efficiently increase the transient expression of recombinant proteins in Nicotiana benthamiana manifold.

Full-Text [PDF 732 kb]   (455 Downloads)    
Type of Study: Research | Subject: Biotechnology
Received: 2016/08/25 | Accepted: 2016/11/5 | Published: 2016/11/5

Add your comments about this article : Your username or email:
Write the security code in the box

Send email to the article author


© 2015 All Rights Reserved | Research in Molecular Medicine

Designed & Developed by : Yektaweb