ABSTRACT
Contents Purified messenger ribonucleic acids (mRNA) from total RNAs or mRNA transcribed in vitro from cloned complementary deoxyribonucleic acid (cDNA) inserts can be translated in cell-free extracts to produce protein(s). The synthesized protein(s) can then be analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis, immunoprecipitation, or biological activity assay, depending on the particular research interest.1-5 The present chapter describes, step by step, the protocols for in vitro translation of mRNA(s) using commercial lysates of rabbit reticulocytes and wheat germ extracts, and protocols for analysis of the products by SDSPAGE. The basic principle is that commercial translational kits contain translational machinery, including all the components such as amino acids, transfer RNA (tRNA), 0-8493-0815-1/04/$0.00+$ 1.50
ribosomal RNA (rRNA), and factors (initiation, elongation, and termination) for the in vitro synthesis of either radioactively labeled or unlabeled polypeptide(s).
