Abstract
in vitrotranslation is a classical method of molecular biology (Clemens, 1984). All three phases of the cellular translational process - initiation, elongation, and termination - can be reconstituted in the test tube. Defined natural RNAs or RNAs synthesizedin vitro(Craig et al., 1992; Krieg and Melton, 1984) are added as templates to the translation system and program the synthesis of the specific encoded proteins. Different cell free systems capable of efficiently translating a broad variety of exogenously supplied mRNA templates have been described. In most cases it is not necessary to further optimize translation conditions of the mRNA used. Lysates for cell free protein synthesis can be prepared from rabbit reticulocytes (Jackson et al., 1983; Pelham and Jackson, 1976), wheat germ (Erickson and Blobel, 1983; Roberts and Paterson, 1973)E.coli(DeVries and Zubay, 1969; Pratt, 1984; Zubay, 1973)Stapyhlococcus aureusor yeast (Hofbauer et al., 1982; Tuite et al., 1980; Tuite and Plesset, 1986).
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Metzler, T., Hans-Joachim, H. (2000). Labeling of Proteins During In Vitro Translation. In: Kessler, C. (eds) Nonradioactive Analysis of Biomolecules. Springer Lab Manuals. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-57206-7_7
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DOI: https://doi.org/10.1007/978-3-642-57206-7_7
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