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The first three fragments were, respectively subcloned into pRS425 (leu2 auxotrophic marker) digested by Xho I and Hin dIII, resulting in the multi-copy plasmids pVTC2-GLDH, pALO1-VTC2, and pALO1-GLDH

Chemical crosslinking was carried out in a total reaction volume of 1.0 mL containing 4 M of Syn-CSK protein

Once inside the cell, the -glutamyl-amino acid can be metabolized to release the amino acid and 5-oxoproline, which can then be converted into glutamate to be used in the synthesis of GSH