Demonstration of the early--late switch in vitro with bacteriophage T7 DNA as template.
A protein-synthesizing system in vitro, programmed with bacteriophage T7 DNA as template, changed the specificity of gene expression in the course of incubation as a result of newly synthesized T7 early proteins. The system mimics largely the situation in vivo on both the transcriptional and the translational levels, i.e. early gene expression is turned off shortly after late synthesis has been started. These results suggest that the switch from early to late expression does not necessarily require changes in the cellular environment nor is it dependent on the presence of membranes. The main part in this process is played by the phage-dependent RNA polymerase (gene 1 product), whose activity appears 8-10 min after start of incubation. When its activity is reduced by inhibitors or creation of non-optimal conditions, the system is not able to manage the early--late switch.
D004279:DNA, Viral / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D066298:In Vitro Techniques; D009693:Nucleic Acid Hybridization; D012367:RNA, Viral / Q000096:biosynthesis; D013604:T-Phages / Q000378:metabolism*; D013698:Templates, Genetic; D014158:Transcription, Genetic
D004279:DNA, Viral; D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases