Transcription efficiency along the tissue-plasminogen-activator cDNA gene in Escherichia coli.
Transcription along the human tissue-plasminogen activator gene (tpa) cloned in Escherichia coli was studied by inserting the 4.5S RNA gene at various locations within tpa, and measuring the accumulation of 4.5S RNA. A six-fold decrease in transcription of 4.5S RNA was observed as the distance from the Ptrc promoter to the 4.5S RNA gene increased. Independent measurements indicated that the quantity of initiations from the Ptrc promoter could not account for the differences observed. Because the 4.5S RNA is stable, the decrease in transcription was assumed to be due to premature transcription-termination within the tpa gene. This polar effect could contribute to poor expression of tpa in E. coli.
D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004720:Endonucleases; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D006801:Humans; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics; D015719:Single-Strand Specific DNA and RNA Endonucleases; D010959:Tissue Plasminogen Activator / Q000235:genetics*; D014158:Transcription, Genetic
D012313:RNA; D004247:DNA; D004720:Endonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases; D010959:Tissue Plasminogen Activator