Abstract
Fusions of elastin-like peptide (ELP) purification tags and self-cleaving inteins provide a powerful platform for purifying tagless recombinant proteins without the need for conventional packed-bed columns. A drawback to this method has been premature cleaving of the ELP tag during expression, before the purification procedure can take place. Here we demonstrate a split-intein method, where the self-cleaving intein is divided into two inactive segments during expression and purification. Spontaneous assembly of the purified intein segments then restores self-cleaving activity to deliver the tagless target protein.
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Shi, C., Han, TC., Wood, D.W. (2017). Purification of Microbially Expressed Recombinant Proteins via a Dual ELP Split Intein System. In: Mootz, H. (eds) Split Inteins. Methods in Molecular Biology, vol 1495. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6451-2_2
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DOI: https://doi.org/10.1007/978-1-4939-6451-2_2
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Online ISBN: 978-1-4939-6451-2
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