Transformation strategies for stable expression of complex hetero-multimeric proteins like secretory immunoglobulin A in plants.
molecular farming
multimeric glycoprotein
recombinant protein production
secretory IgA
seed specific expression
transgene expression variability
Journal
Plant biotechnology journal
ISSN: 1467-7652
Titre abrégé: Plant Biotechnol J
Pays: England
ID NLM: 101201889
Informations de publication
Date de publication:
09 2019
09 2019
Historique:
received:
16
04
2018
revised:
12
02
2019
accepted:
18
02
2019
pubmed:
26
2
2019
medline:
4
3
2020
entrez:
26
2
2019
Statut:
ppublish
Résumé
Plant expression systems have proven to be exceptional in producing high-value complex polymeric proteins such as secretory IgAs (SIgAs). However, polymeric protein production requires the expression of multiple genes, which can be transformed as single or multiple T-DNA units to generate stable transgenic plant lines. Here, we evaluated four strategies to stably transform multiple genes and to obtain high expression of all components. Using the in-seed expression of a simplified secretory IgA (sSIgA) as a reference molecule, we conclude that it is better to spread the genes over two T-DNAs than to contain them in a single T-DNA, because of the presence of homologous recombination events and gene silencing. These T-DNAs can be cotransformed to obtain transgenic plants in one transformation step. However, if time permits, more transformants with high production levels of the polymeric protein can be obtained either by sequential transformation or by in-parallel transformation followed by crossing of transformants independently selected for excellent expression of the genes in each T-DNA.
Identifiants
pubmed: 30801876
doi: 10.1111/pbi.13098
pmc: PMC6686127
doi:
Substances chimiques
DNA, Bacterial
0
Immunoglobulin A, Secretory
0
T-DNA
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1760-1769Informations de copyright
© 2019 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.
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