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. 2023 Nov 29;15(12):2344. doi: 10.3390/v15122344

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© 2023 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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Detection of different products of autoproteolysis of His-Prot-Pol*. Total protein of non-induced (n.i.) and induced (i.) cultures of E. coli transformed with plasmid pet11a-His-Prot-Pol* were resolved in SDS-PAGE and analyzed via Western blot. (A) Mab 10B6 anti-His showed the full-length His-Prot-Pol* (60 kDa) and the histidine-tagged cleavage product His-Prot (35 kDa). Again, a weak 25 kDa band (marked with an arrowhead) appeared, which was not further characterized. (B) Mab 55A10 anti-Pol reacted with the 60 kDa His-Prot-Pol* and two smaller fragments of 26 and 28 kDa representing the released Pol* moiety and an uncharacterized fragment. (C) Mab 55B20 anti-Prot in contrast showed a pattern similar to the His-tag specific Mab with bands at 60 and 35 kDa. His-Prot-Pol*, His-Prot, and Pol* fragments are marked with arrows.