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Are proteins produced by an in vitro transcription/ translation system using Transcend™ tRNA functional?

The Transcend™ tRNA is a precharged ε-labeled biotinylated lysine-tRNA complex that allows nonradioactive detection of proteins produced by in vitro translation. When using Transcend™ tRNA, approximately 25–33% of lysine residues incorporated into the protein are biotinylated lysines. At Promega, we have evaulated three proteins for enzymatic activity. Both chloramphenicol acetyltransferase and firefly luciferase are fully active when biotinylated lysines are incorporated. This is primarily due to the fact that none of the lysines present in the proteins are located in the active sites. However, beta-galactosidase activity was decreased. The modified lysines residues may interfere with tetramer formation, which is required for enzyme activity.