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  • Title: A Neurospora crassa heat-shocked cell lysate translates homologous and heterologous messenger RNA efficiently, without preference for heat shock messages.
    Author: Curle CA, Kapoor M.
    Journal: Curr Genet; 1988 May; 13(5):401-9. PubMed ID: 2969781.
    Abstract:
    Cell-free protein synthesis systems were prepared from normally-grown (N-lysate) and heat-shocked (HS-lysate) Neurospora crassa mycelium. Although both lysates translated homologous mRNA, the HS-lysate was more active, yielding a higher incorporation of [35S]-methionine into hot TCA-insoluble material and a vastly superior protein synthesis profile. The optimal temperature for translation by both lysates was 21 degrees C; the HS-lysate did not translate heat-shock mRNA preferentially at any temperature tested. Fortuitously, heterologous messenger RNAs from diverse eukaryotic and viral sources - Drosophila, dog pancreas, rabbit globin mRNA, brome mosaic virus, tobacco mosaic virus - were translated by the HS-lysate with an efficiency comparable to that of the commercial rabbit reticulocyte system and superior to the wheat germ system. The cap analogues, m7G(5')ppp(5')G and m7G(5')Gm, inhibited translation significantly.
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