Fig. 1 in Rapid screening of glycosyltransferases in plants using a linear DNA expression template based cell-free transcription-translation system

Fig. 1. Rapid screening of plant glycosyltransferases using the LET-based-TX-TL system. We can either use long primers which contain a promoter, a ribosome binding site, and a terminator to generate expressible linear DNAs or use short primers to amplify the targeted gene fragments and then ligate them with a promoter, a ribosome binding site, a terminator, and a backbone; then another pair of primers is used to generate expressible linear DNAs. Afterward, combine TX-TL extracts, buffers, and expressible linear DNAs to start protein expression. Then this TX-TL mixture is directly added with substrates (such as quercetin) to start glycosylation reactions. Finally, UPLC-MS is used to analyze the reaction mixture to examine whether targeted products (such as isoquercitrin) are generated.