Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield

Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression:... Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Horticulture, Environment, and Biotechnology Springer Journals

Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield

Loading next page...
 
/lp/springer_journal/agroinfiltration-of-leaves-for-deconstructed-viral-vector-based-WslftUVYIV
Publisher
Springer Journals
Copyright
Copyright © 2018 by Korean Society for Horticultural Science and Springer-Verlag GmbH Germany, part of Springer Nature
Subject
Life Sciences; Life Sciences, general; Plant Breeding/Biotechnology; Plant Physiology; Agriculture; Plant Ecology
ISSN
2211-3452
eISSN
2211-3460
D.O.I.
10.1007/s13580-018-0047-6
Publisher site
See Article on Publisher Site

Abstract

Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration.

Journal

Horticulture, Environment, and BiotechnologySpringer Journals

Published: Jun 5, 2018

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create lists to
organize your research

Export lists, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off