Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I-1-I-2 regulatory elements

被引:1271
作者
Lutz, R [1 ]
Bujard, H [1 ]
机构
[1] UNIV HEIDELBERG, ZENTRUM MOL BIOL, D-69120 HEIDELBERG, GERMANY
关键词
D O I
10.1093/nar/25.6.1203
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Based on parameters governing promoter activity and using regulatory elements of the lac, ara and tet operon transcription control sequences were composed which permit the regulation in Escherichia coli of several gene activities independently and quantitatively. The novel promoter PLtetO-1 allows the regulation of gene expression over an up to 5000-fold range with anhydrotetracycline (aTc) whereas with IPTG and arabinose the activity of Plac/ara-1 may be controlled 1800-fold. Escherichia coli host strains which produce defined amounts of the regulatory proteins, Lac and Tet repressor as well as AraC from chromosomally located expression units provide highly reproducible in vivo conditions, Controlling the expression of the genes encoding luciferase, the low abundance E.coli protein DnaJ and restriction endonuclease Cfr9I not only demonstrates that high levels of expression can be achieved but also suggests that under conditions of optimal repression only around one mRNA every 3rd generation is produced, This potential of quantitative control will open up new approaches in the study of gene function in vivo, in particular with low abundance regulatory gene products, The system will also provide new opportunities for the controlled expression of heterologous genes.
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页码:1203 / 1210
页数:8
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