Journal: The Journal of Biological Chemistry
Article Title: Selenocysteine Insertion at a Predefined UAG Codon in a Release Factor 1 (RF1)-depleted Escherichia coli Host Strain Bypasses Species Barriers in Recombinant Selenoprotein Translation *
Figure Lengend Snippet: TrxR1 produced using UAG for Sec in absence of both RF1 and a SECIS element resulted in a mixture of Sec-containing enzyme, UAG-truncated variant, and additional forms presumed to be Sec-to-Gln or Sec-to-Lys substituted variants. Shown is an electrospray mass spectrometry analysis of rat TrxR1 produced using the pABC2-rTR UAG plasmid in C321.ΔA host cells. The three major species of enzyme detected corresponded in size to a native Sec-containing variant ( green arrow ), UAG-truncated enzyme ( red arrow ), and a variant likely corresponding to Sec-to-Gln- or Sec-to-Lys-substituted forms ( blue arrow ). See text for further discussion.
Article Snippet: The gene for human TrxR1 was synthesized by a commercial service (GeneArt, ) with codons optimized for expression in E. coli and cloned into the pABC2 vector between XbaI and EagI, which resulted in plasmid pABC2-hTRSUAG .
Techniques: Produced, Size-exclusion Chromatography, Variant Assay, Mass Spectrometry, Plasmid Preparation