Multicolor in vitro translation.

Published

Journal Article

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 by detection of labeled amino acids incorporated during translation. The original strategy of incorporating radioactively labeled amino acids, such as [35S]methionine or [14C]leucine, has been superseded by the addition of antigenic tags or the incorporation of biotin-labeled or BODIPY-FL-labeled amino acids. Such nonradioactive tags are easier to visualize after translation and do not pose a radiation hazard. Among the nonradioactive tags, BODIPY-FL-lysine offers the advantage that proteins that have incorporated this amino acid can be directly visualized after gel electrophoresis. We show here that multiple fluorophores introduced into proteins can considerably extend their usefulness, particularly for the comparison of in vitro-translated proteins from related sources. This technology can be applied in various situations, including the simplified detection of rare truncating mutations in clinical samples from cancer patients.

Full Text

Duke Authors

Cited Authors

  • Traverso, G; Diehl, F; Hurst, R; Shuber, A; Whitney, D; Johnson, C; Levin, B; Kinzler, KW; Vogelstein, B

Published Date

  • September 2003

Published In

Volume / Issue

  • 21 / 9

Start / End Page

  • 1093 - 1097

PubMed ID

  • 12910244

Pubmed Central ID

  • 12910244

Electronic International Standard Serial Number (EISSN)

  • 1546-1696

International Standard Serial Number (ISSN)

  • 1087-0156

Digital Object Identifier (DOI)

  • 10.1038/nbt857

Language

  • eng