Preparation of a cell-free translation system from a wild-type strain of Neurospora crassa and translation of pyruvate kinase messenger RNA.
A cell-free in vitro translation system exhibiting high activity has been developed from wild-type Neurospora crassa mycelium. The isolation is simple and fast, and the homogenization does not appear to affect the activity of mycelial proteases and nucleases. This system is capable of supporting efficient translation of exogenously added homologous RNA as demonstrated by the experiments with PK-specific mRNA. In addition, it translates heterologous RNA efficiently, shown by the translation of globin mRNA. We did not examine the Neurospora lysate for post-translational modification activity. The procedure used for the preparation of Neurospora cell-free extracts should be readily applicable to the other filamentous fungi.
D002474:Cell-Free System; D007700:Kinetics; D008836:Micrococcal Nuclease; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology* / Q000235:genetics; D014176:Protein Biosynthesis; D011770:Pyruvate Kinase / Q000235:genetics*; D012331:RNA, Fungal / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics*
D012331:RNA, Fungal; D012333:RNA, Messenger; D011770:Pyruvate Kinase; D008836:Micrococcal Nuclease