| 简介 |
In vitro translation is a widely used tool for both analytical and preparative purposes. For analytical purposes, small amounts of proteins are synthesized and visualized through labeled amino acids incorporated during translation. The radioactively labeled amino acids originally used, such as [S]methionine or [C]leucine, have been superceded by the addition of antigenic tags or the incorporation of biotin-labeled or fluorescently-labeled amino acids. Such non-radioactive tags are simpler to visualize following translation and do not pose a radiation hazard. Among the non-radioactive tags, fluorescently labeled-lysine is the most convenient, as proteins that have incorporated this amino acid can be directly visualized following gel electrophoresis. Multiple fluorophores introduced into proteins significantly extend their utility, particularly for the comparison of in vitro translated proteins from related sources. This methodology can be employed for several purposes, including the simplified detection of rare truncating mutations in clinical samples from cancer patients. |
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