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Protein Coexpression Using FMDV 2A: Effect of “Linker” Residues
Joint Authors
Minskaia, Ekaterina
Ryan, Martin D.
Source
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-12, 12 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-06-11
Country of Publication
Egypt
No. of Pages
12
Main Subjects
Abstract EN
Many biomedical applications absolutely require, or are substantially enhanced by, coexpression of multiple proteins from a single vector.
Foot-and-mouth disease virus 2A (F2A) and “2A-like” sequences (e.g., Thosea asigna virus 2A; T2A) are used widely for this purpose since multiple proteins can be coexpressed by linking open reading frames (ORFs) to form a single cistron.
The activity of F2A “cleavage” may, however, be compromised by both the use of shorter versions of F2A and the sequences (derived from multiple-purpose cloning sites) used to link F2A to the upstream protein.
To characterise these effects, different lengths of F2A and T2A were inserted between green and cherry fluorescent proteins.
Mutations were introduced in the linker region immediately upstream of both F2A- and T2A-based constructs and activities determined using both cell-free translation systems and transfected cells.
In shorter versions of F2A, activity may be affected by both the C-terminal sequence of the protein upstream and, equally strikingly, the residues immediately upstream introduced during cloning.
Mutations significantly improved activity for shorter versions of F2A but could decrease activity in the case of T2A.
These data will aid the design of cloning strategies for the co-expression of multiple proteins in biomedical/biotechnological applications.
American Psychological Association (APA)
Minskaia, Ekaterina& Ryan, Martin D.. 2013. Protein Coexpression Using FMDV 2A: Effect of “Linker” Residues. BioMed Research International،Vol. 2013, no. 2013, pp.1-12.
https://search.emarefa.net/detail/BIM-1003913
Modern Language Association (MLA)
Minskaia, Ekaterina& Ryan, Martin D.. Protein Coexpression Using FMDV 2A: Effect of “Linker” Residues. BioMed Research International No. 2013 (2013), pp.1-12.
https://search.emarefa.net/detail/BIM-1003913
American Medical Association (AMA)
Minskaia, Ekaterina& Ryan, Martin D.. Protein Coexpression Using FMDV 2A: Effect of “Linker” Residues. BioMed Research International. 2013. Vol. 2013, no. 2013, pp.1-12.
https://search.emarefa.net/detail/BIM-1003913
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1003913