By Herbert Weissbach
Dimensions In health and wellbeing examine: look for The medications Of Tomorrow
summary: Dimensions In health and wellbeing learn: look for The medications Of the following day
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Extra resources for Dimensions in Health Research: Search for the Medicines of Tomorrow
RNA was extracted from the induced cells and the polyadenylated mRNA was isolated from the total cellular RNA by fractionation on oligo(dT)-cellulose. Translation of the mRNA was performed in an Ehrlich ascites cell-free extract (mouse S-30), The products of translation were assayed directly for interferon activity (11, 16). the 60-min incubation at 30° (Table II). The data show that only reaction mixtures incubated at 30° for 60 min yielded interferon activity. In addition, sparsomycin, a specific inhibitor of protein synthesis (13), abolished the synthesis of human interferon in these cell-free extracts (Table II).
The results of these studies (12) are summarized below. Synthesis of Human Interferon by Xenopus laevis Oocytes. Polyadenylated mRNA from induced FS-4 fibroblasts was injected into Xenopus laevis oocytes. Synthesis of human interferon as a function of time at three different mRNA concentrations is shown in Figure 2. At 7 ng and 39 ng of mRNA per oocyte, synthesis of interferon was linear for at least six hours. At 39 HUMAN INTERFERON o X O 16 24 TIME (hours) FIGURE 2. Interferon synthesis as a function of time.
In this case, the same polypeptide sequence would be synthesized, but processed differently into the final active product in the different types of cells. To distinguish between these alternatives, we isolated mRNA from human lymphoblasts and fibroblasts induced to produce their characteristic interferons? and injected the mRNA into Xenopus laevis oocytes. Xenopus oocytes have been shown to translate a variety of injected mRNAs and to carry out a number of post-translational modifications including polypeptide cleavage (30, 31), acetylation (32), and glycosylation (33).