FlexiBAC: a versatile, open-source baculovirus vector system for protein expression, secretion, and proteolytic processing.

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  • Additional Information
    • Source:
      Publisher: BioMed Central Country of Publication: England NLM ID: 101088663 Publication Model: Electronic Cited Medium: Internet ISSN: 1472-6750 (Electronic) Linking ISSN: 14726750 NLM ISO Abbreviation: BMC Biotechnol
    • Publication Information:
      Original Publication: [London] : BioMed Central, 2001-
    • Subject Terms:
      Protein Processing, Post-Translational*; Baculoviridae/*genetics ; Genetic Vectors/*genetics ; Recombinant Proteins/*genetics; Activins/biosynthesis ; Activins/genetics ; Animals ; Gene Expression ; Recombinant Proteins/metabolism ; Spodoptera/genetics ; Transfection/methods ; Virus Cultivation/methods
    • Abstract:
      Background: Baculovirus-mediated expression in insect cells is a powerful approach for protein production. However, many existing methods are time-consuming, offer limited options for protein tagging, and are unsuitable for secreted proteins requiring proteolytic maturation, such as TGF-β family growth factors.
      Results: To overcome the limitations of traditional baculovirus expression systems, we engineered "FlexiBAC". This system allows recombinant baculovirus formation inside insect cells and reduces the time between initial cloning and protein production to 13 days. FlexiBAC includes 143 shuttle vectors that append combinations of purification tags, fluorescent markers, proteolytic cleavage sites, trafficking signals, and chemical conjugation tags to the termini of the target protein. This system also overexpresses recombinant furin convertase to allow efficient proteolytic processing of secreted proteins. We demonstrate that FlexiBAC can be used to produce high levels of mature, active forms of TGF-β family growth factors, such as Activin A, as well as other proteins that are typically difficult to reconstitute, such as proteins rich in coiled-coil, low complexity, and disordered domains.
      Conclusions: FlexiBAC is a protein expression system for production of both cytosolic proteins and secreted proteins that require proteolytic maturation. The design of FlexiBAC and its expansive complementary shuttle vector system reduces cloning steps and simplifies baculovirus production.
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    • Contributed Indexing:
      Keywords: Baculovirus; Intrinsically disordered proteins; Protein expression; TGF-β
    • Accession Number:
      0 (Recombinant Proteins)
      0 (activin A)
      104625-48-1 (Activins)
    • Publication Date:
      Date Created: 20190331 Date Completed: 20190730 Latest Revision: 20200225
    • Publication Date:
      20240829
    • Accession Number:
      PMC6441187
    • Accession Number:
      10.1186/s12896-019-0512-z
    • Accession Number:
      30925874