Abstract
In gram-negative bacteria, assembly of outer membrane proteins requires the multicomponent β-barrel assembly machinery (BAM) complex, of which BamA is an essential and evolutionarily conserved integral outer membrane protein. To understand how BamA facilitates outer membrane protein biogenesis, it is important to obtain sufficient amounts of purified recombinant BamA protein for in vitro functional analysis and structure determination. In this chapter, we describe the protocol that we used in our laboratory for the cloning, expression, and purification of E. coli BamA and its N-terminal deletion variants for in vitro functional studies and for structure determination of the β-barrel domain alone (residues 426–810).
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Acknowledgements
The work was supported by grants from the Ministry of Science and Technology and the National Natural Science Foundation of China.
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Ni, D., Huang, Y. (2015). The Expression, Purification, and Structure Determination of BamA from E. coli . In: Buchanan, S., Noinaj, N. (eds) The BAM Complex. Methods in Molecular Biology, vol 1329. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2871-2_13
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DOI: https://doi.org/10.1007/978-1-4939-2871-2_13
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2870-5
Online ISBN: 978-1-4939-2871-2
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