A vector design that allows fast and convenient production of differently tagged proteins

Omar Scapolan, Andrea N. Mazzarello, Maria Bono, Marzia Occhino, Vasily Ogryzko, Marco Bestagno, Paolo Scartezzini, Silvia Bruno, Franco Fais, Fabio Ghiotto

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Recombinant-tagged proteins have a widespread use in experimental research as well as in clinical diagnostic and therapeutic approaches. Well-stocked sets of differently tagged variants of a same protein would be of great help. However, the construction of differently tagging vectors is a demanding task since cloning procedures need several tailored DNA inserts. In this study, we describe a novel vector system that allows a cost- and time-effective production of differently tagged variants of a same protein by using the same DNA fragment and a set of vectors each carrying a different tag. The design of these expression vectors is based on an intronic region that becomes functional upon cloning the insert sequence, splicing of which attaches a certain tag to the protein termini. This strategy allows for the cloning of the fragment that codes for the protein of interest, without any further modification, into different vectors, previously built and ready-to-use, each carrying a tag that will be joined to the protein. Proof of principle for our expression system, presented here, is shown through the production of a functional anti-GD2 Fab fragment tagged with biotin or polyhistidine, or a combination of both, followed by the demonstration of the functional competencies of both the protein and the tags.

Original languageEnglish
Pages (from-to)16-25
Number of pages10
JournalMolecular Biotechnology
Volume52
Issue number1
DOIs
Publication statusPublished - Sep 2012

Fingerprint

Proteins
Cloning
Organism Cloning
DNA
Recombinant proteins
Immunoglobulin Fab Fragments
Biotin
Recombinant Proteins
Demonstrations
Costs and Cost Analysis
Research
Costs
Therapeutics

Keywords

  • Expression vector
  • Fab fragment
  • Immunotherapy
  • Protein tag
  • Recombinant antibody

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Molecular Biology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Scapolan, O., Mazzarello, A. N., Bono, M., Occhino, M., Ogryzko, V., Bestagno, M., ... Ghiotto, F. (2012). A vector design that allows fast and convenient production of differently tagged proteins. Molecular Biotechnology, 52(1), 16-25. https://doi.org/10.1007/s12033-011-9469-4

A vector design that allows fast and convenient production of differently tagged proteins. / Scapolan, Omar; Mazzarello, Andrea N.; Bono, Maria; Occhino, Marzia; Ogryzko, Vasily; Bestagno, Marco; Scartezzini, Paolo; Bruno, Silvia; Fais, Franco; Ghiotto, Fabio.

In: Molecular Biotechnology, Vol. 52, No. 1, 09.2012, p. 16-25.

Research output: Contribution to journalArticle

Scapolan, O, Mazzarello, AN, Bono, M, Occhino, M, Ogryzko, V, Bestagno, M, Scartezzini, P, Bruno, S, Fais, F & Ghiotto, F 2012, 'A vector design that allows fast and convenient production of differently tagged proteins', Molecular Biotechnology, vol. 52, no. 1, pp. 16-25. https://doi.org/10.1007/s12033-011-9469-4
Scapolan, Omar ; Mazzarello, Andrea N. ; Bono, Maria ; Occhino, Marzia ; Ogryzko, Vasily ; Bestagno, Marco ; Scartezzini, Paolo ; Bruno, Silvia ; Fais, Franco ; Ghiotto, Fabio. / A vector design that allows fast and convenient production of differently tagged proteins. In: Molecular Biotechnology. 2012 ; Vol. 52, No. 1. pp. 16-25.
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