A Dual-Intein Autoprocessing Domain that Directs Synchronized Protein Co-Expression in Both Prokaryotes and Eukaryotes

Being able to coordinate co-expression of multiple proteins is necessary for a variety of important applications such as assembly of protein complexes, trait stacking and metabolic engineering. Currently only few options are available for multiple recombinant protein co-expression and most of them a...

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Veröffentlicht in:Scientific reports 2015-02, Vol.5 (1), p.8541-8541, Article 8541
Hauptverfasser: Zhang, Bei, Rapolu, Madhusudhan, Liang, Zhibin, Han, Zhenlin, Williams, Philip G., Su, Wei Wen
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Sprache:eng
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Zusammenfassung:Being able to coordinate co-expression of multiple proteins is necessary for a variety of important applications such as assembly of protein complexes, trait stacking and metabolic engineering. Currently only few options are available for multiple recombinant protein co-expression and most of them are not applicable to both prokaryotic and eukaryotic hosts. Here, we report a new polyprotein vector system that is based on a pair of self-excising mini-inteins fused in tandem, termed the dual-intein (DI) domain, to achieve synchronized co-expression of multiple proteins. The DI domain comprises an Ssp DnaE mini-intein N159A mutant and an Ssp DnaB mini-intein C1A mutant connected in tandem by a peptide linker to mediate efficient release of the flanking proteins via autocatalytic cleavage. Essentially complete release of constituent proteins, GFP and RFP (mCherry), from a polyprotein precursor, in bacterial, mammalian and plant hosts was demonstrated. In addition, successful co-expression of GFP with chloramphenicol acetyltransferase and thioredoxin with RFP, respectively, further substantiates the general applicability of the DI polyprotein system. Collectively, our results demonstrate the DI-based polyprotein technology as a highly valuable addition to the molecular toolbox for multi-protein co-expression which finds vast applications in biotechnology, biosciences and biomedicine.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep08541