flashBAC Publications

Examples of flashBAC usage in recent publications:

As an innovative, research-driven company with a strong scientific base, we are always very pleased to see the flashBAC system being used in scientific research that has appeared in the literature. Here we present a selection of highly interesting recently published papers where the flashBAC system has been used. These papers demonstrate how the system has been used to express a variety of proteins (including virus proteins, ion channels and ribosomal proteins) for a diverse range of purposes including structural biology, immunology and the study of phosphorylation patterns.

1. Chiam R, Sharp E, Maan S, Rao S, Mertens P, Blacklaws B, Davis-Poynter N, Wood J, Castillo-Olivares J. Induction of antibody responses to African horse sickness virus (AHSV) in ponies after vaccination with recombinant modified vaccinia Ankara (MVA). PLoS One. 2009 Jun 22;4(6):e5997. PubMed PMID: 19543394.

   In this paper, Chiam et al. used the flashBAC system to express recombinant modified vaccinia Ankara (MVA) proteins. The flashBAC expression system was used in order to obtain a source of antigens for use in immunological assays. The VP2, VP7 and NS3 genes of AHSV were expressed and the protein obtained used in this study to investigating the ability of recombinant MVA to stimulate immune responses against AHSV antigens in the horse.

2. Enuka Y, Hanukoglu I, Edelheit O, Vaknine H, Hanukoglu A. Epithelial sodium channels (ENaC) are uniformly distributed on motile cilia in the oviduct and the respiratory airways. Histochem Cell Biol. 2012 Mar;137(3):339-53. Epub 2011 Dec 30. PubMed PMID: 22207244.

   Here Enuka et al. employed the flashBAC system to generate the full length alpha, beta, and gamma subunits of the human epithelial sodium channel.

3. Rozen-Gagnon K, Moreland NJ, Ruedl C, Vasudevan SG. Expression and immunoaffinity purification of recombinant dengue virus 2 NS1 protein as a cleavable SUMOstar fusion. Protein Expr Purif. 2012 Mar;82(1):20-5. Epub 2011 Nov 11. PubMed PMID: 22100526.

   Here flashBAC was used by Rozen-Gagnon et al. to produce a cleavable SUMOStar tagged NS1 protein of Dengue virus 2 and were able to obtain milligram quantities of their protein of interest. Commenting on their choice of system, these authors commented “Unlike native NS1 purification from infected mammalian cells, the baculovirus system is readily amenable to mutagenesis and other protein manipulations making rNS1 a valuable tool for studying dengue NS1 functionality."

4. Brunotte L, Kerber R, Shang W, Hauer F, Hass M, Gabriel M, Lelke M, Busch C, Stark H, Svergun DI, Betzel C, Perbandt M, Günther S. Structure of the Lassa virus nucleoprotein revealed by X-ray crystallography, small-angle X-ray scattering, and electron microscopy. J Biol Chem. 2011 Nov 4;286(44):38748-56. Epub 2011 Sep 14. PubMed PMID: 21917929.

   The nucleoprotein of Lassa Virus was expressed using the flashBAC system for use in full length crystal structure studies. This allowed this group to determine the crystal and quaternary structures of the Lassa virus NP and show that Lassa virus NP assembles into a symmetric trimer in solution. The trimeric complex may have a biological function in the virus life cycle, and its assembly could be a target for antivirals.

5. Ramya R, Mohana Subramanian B, Sivakumar V, Senthilkumar RL, Sambasiva Rao KR, Srinivasan VA. Expression and solubilization of insect cell-based rabies virus glycoprotein and assessment of its immunogenicity and protective efficacy in mice. Clin Vaccine Immunol. 2011 Oct;18(10):1673-9. Epub 2011 Aug 3. PubMed PMID: 21813661.

   In this paper Ramya et al. used the flashBAC system to express the Rabies Virus Glycoprotein for use in their studies into solubilising the protein for use in potential vaccines. They demonstrated that when subjected to the appropriate solubilisation protocol, the recombinant proteins retained the immunologically relevant domains in the native conformation, demonstrating their potential for use in a novel subunit vaccine.

6. Sunami T, Byrne N, Diehl RE, Funabashi K, Hall DL, Ikuta M, Patel SB, Shipman JM, Smith RF, Takahashi I, Zugay-Murphy J, Iwasawa Y, Lumb KJ, Munshi SK, Sharma S. Structural basis of human p70 ribosomal S6 kinase-1 regulation by activation loop phosphorylation. J Biol Chem. 2010 Feb 12;285(7):4587-94. Epub 2009 Oct 28. PubMed PMID: 19864428.

   Here Sunami et al. used the flashBAC system to express Human p70 ribosomal s6 kinase 1for use in structure phosphorylation studies. This was the first structural description of the p70S6K1 kinase domain in both its inactive and active forms. These data offer insight into potential directions for structure guided design of specific p70s6k inhibitors.