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. 2021 Aug 23;11(8):2144. doi: 10.3390/nano11082144

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Construction, expression and purification of recombinant proteins. (A) Schematic representation of the reading frame of the expression vector TRX-hβD2 and TRX-hβD2-M, (B) expression and purification of hβD2; lane 1: molecular weight marker, lane 2: insoluble fraction, lane 3: soluble fraction, lane 4: fraction not bound to the first Ni²⁺-IMAC purification step, lane 5: fraction bound to the first Ni²⁺-IMAC purification step, lane 6: fraction after cleavage by TEV protease, lane 7: fraction bound to the second purification step by Ni²⁺-IMAC and lane 8: fraction not bound to the second purification step by Ni²⁺-IMAC. (C) expression and purification of hβD2-M; the charging plan is the same as for hβD2.