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biGBac enables rapid gene assembly for the expression of large multisubunit protein complexes
Open accessAbstract
Significance: At the molecular level, most processes in living systems are mediated by multisubunit protein complexes. Recombinant forms of these complexes are essential for analyzing their structure and function. Multigene expression constructs greatly improve recombinant protein complex preparations, but the generation of such constructs can be a rate-limiting step. To overcome this limitation, we have adapted Gibson assembly reactions for the rapid, efficient, and fast generation of numerous expression constructs in parallel and used the resulting biGBac method for expression of different cell-cycle complexes, composed of up to 17 different subunits. The biGBac technique enables the analyses of large protein complexes by systematic mutagenesis approaches that were not feasible before.
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Self-citationStructural and mechanistic basis of the EMC-dependent biogenesis of distinct transmembrane clients
Miller-Vedam, L. E., Bräuning, B., Popova, K. D., Schirle Oakdale, N. T., Bonnar, J. L., Prabu, J. R., Boydston, E. A., Sevillano, N., Shurtleff, M. J., Stroud, R. M., et al. (2020) · eLife · doi:10.7554/elife.62611 · Cited by 87
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Metadata
- Authors
- Weissmann, F., Petzold, G., VanderLinden, R., Huis in 't Veld, P. J., Brown, N. G., Lampert, F., Westermann, S., Stark, H., Schulman, B. A., & Peters, J.-M.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2016-04-25
- Volume
- 113
- Issue
- 19
- Pages
- E2564-9
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2344664973
- Cited by
- 355
- References
- 0
- FWCI
- 6.77
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Weissmann, F., Petzold, G., VanderLinden, R., Huis in 't Veld, P. J., Brown, N. G., Lampert, F., Westermann, S., Stark, H., Schulman, B. A., & Peters, J.-M. (2016). biGBac enables rapid gene assembly for the expression of large multisubunit protein complexes. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.1604935113
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