Research in Molecular Medicine
Research in Molecular Medicine
Res Mol Med (RMM)
Medical Sciences
http://rmm.mazums.ac.ir
1
admin
2322-1348
2322-133X
10.29252/rmm
en
jalali
1395
5
1
gregorian
2016
8
1
4
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online
1
fulltext
en
Utility of P19 Gene-Silencing Suppressor for High Level Expression of Recombinant Human Therapeutic Proteins in Plant Cells
بیوتکنولوژی
Biotechnology
پژوهشي
Research
<p style="text-align: justify;"><strong>Background: </strong>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 <em>Nicotiana benthamiana.</em></p>
<p style="text-align: justify;"><strong>Materials and Methods: </strong>The p19 coding region was inserted in the pCAMBIA using NcoI and BstEII recognition sites. Also, the cloned synthesized recombinant human <em>NGF (rhNGF)</em> 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.</p>
<p style="text-align: justify;"><strong>Results: </strong>Co-agroinfiltration of <em>hNGF</em> with <em>P19</em> suppressor showed about forty-fold increase (8% total soluble protein (TSP)) when compared to the absence of P19 suppressor (0.2%TSP).</p>
<p style="text-align: justify;"><strong>Conclusion:</strong> 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 <em>Nicotiana benthamiana</em> manifold.</p>
Nerve growth Factor, Nicotiana benthamiana, p19 suppressor of gene-silencing, Recombinant therapeutic human protein, Tomato bushy stunt virus.
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http://rmm.mazums.ac.ir/browse.php?a_code=A-10-837-1&slc_lang=en&sid=1
Maryam
Zangi
zangi.m@irost.ir
10031947532846005369
10031947532846005369
No
Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran.
Hamideh
Ofoghi
ofoghi@irost.ir
10031947532846005370
10031947532846005370
Yes
Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran.
Zahra
Amini-Bayat
Amini-Bayat@irost.ir
10031947532846005371
10031947532846005371
No
Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran.
Parastoo
Ehsani
P_ehsani@pasteur.ac.ir
10031947532846005372
10031947532846005372
No
Molecular Biology Unit, Pasteur Institute of Iran, Tehran, Iran.