Abstract
Publisher Summary This chapter describes the translation of HeLa cell histone mRNAs in several in vitro systems and a method for titrating the amount of translatable histone mRNAs in a rabbit reticulocyte lysate. The methods described can also be used to identify histone mRNAs from other sources. HeLa cells are harvested by centrifugation, washed twice with wash buffer, and suspended in lysis buffer at a concentration. In vitro protein synthesizing systems from eukaryotic cells have been successfully used in studies to elucidate the basic mechanism of protein synthesis and to identify specific mRNAs by their translational products. Moreover, in vitro systems that accurately translate mammalian mRNAs are extremely useful to investigate the regulation of m RNA translation and to quantitate biologically active mRNAs from whole cells or from cellular subfractions by their translational capacity.
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