Curle C A, Kapoor M
Department of Biological Sciences, University of Calgary, Alberta, Canada.
Curr Genet. 1988 May;13(5):401-9. doi: 10.1007/BF00365661.
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.
从正常生长的（N-裂解物）和热激的（HS-裂解物）粗糙脉孢菌菌丝体制备无细胞蛋白质合成系统。虽然两种裂解物都能翻译同源mRNA，但HS-裂解物活性更高，[35S]-甲硫氨酸掺入热三氯乙酸不溶性物质的量更高，蛋白质合成谱也明显更优。两种裂解物翻译的最适温度均为21℃；在任何测试温度下，HS-裂解物都不会优先翻译热激mRNA。幸运的是，来自不同真核生物和病毒来源的异源信使RNA——果蝇、犬胰腺、兔珠蛋白mRNA、雀麦花叶病毒、烟草花叶病毒——在HS-裂解物中的翻译效率与商业兔网织红细胞系统相当，且优于小麦胚芽系统。帽类似物m7G(5')ppp(5')G和m7G(5')Gm显著抑制翻译。
