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. 2015 Nov 2;20(11):19735–19747. doi: 10.3390/molecules201119653

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© 2015 by the authors.

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

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Construction of CLK1/pFastBac1 plasmid and the protein expression of CLK1 in insect cells. (A) A schematic of the steps to construct CLK1/pFastBac1 recombinant plasmid; (B) Identification of the constructed CLK1/pFastBac1 recombinant plasmid by both colony PCR assay and double digestion. C-1~5: clone 1 to 5. Cpos: positive clone. Cneg: negative clone; (C) Identification of recombinant bacmid CLK1/pFastBac1 by PCR analysis. NTC: non-template control; (D) Determination the optimal conditions to express the recombinant protein His₆-CLK1 by Coomassie Brilliant Blue staining.