ANeurospora crassaheat-shocked cell lysate translates homologous and heterologous messenger RNA efficiently, without preference for heat shock messages
✍ Scribed by C. A. Curle; M. Kapoor
- Book ID
- 104741894
- Publisher
- Springer-Verlag
- Year
- 1988
- Tongue
- English
- Weight
- 840 KB
- Volume
- 13
- Category
- Article
- ISSN
- 0172-8083
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✦ Synopsis
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 [3Ss]methionine into hot TCA-insoluble material and a vastly superior protein synthesis profile. The optimal temperature for translation by both lysates was 21 °C; the HSlysate 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 mTG(5')ppp(5')Gm, inhibited translation significantly.