T7 infection-dependent selective expression of cloned genes in P1-lysogenic Escherichia coli.
Expression systems based on the selective transcription of genes cloned behind a T7 promoter, by T7 RNA polymerase, display a non-negligible basal expression when the T7 RNA polymerase gene is present within the host organism before induction of the system. This is a problem, especially for cloning and controlled expression of genes toxic to the host organism. We have circumvented this problem by taking advantage of abortive T7 infection of E. coli (P1), in the course of which T7 RNA polymerase is synthesized but bacterial growth is not quantitatively impaired. We have tested this system with three reporter genes, the 6-phospho-beta-galactosidase gene of Staphylococcus aureus, the luciferase operon of Vibrio harveyi, and the rabbit beta-globin gene; we have found very low basal levels, while, upon T7 infection, transcription is at least as efficient as in other in vivo T7 RNA polymerase systems in use.
Institut für Biologie III, Universität Freiburg, FRG.
C
C
Olenik
E
E
Krüger
F
F
Söling
W
W
Hahn
M
M
Messerschmid
R
R
Hausmann
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D005798:Genes, Bacterial; D005914:Globins / Q000235:genetics; D006026:Glycoside Hydrolases; D007700:Kinetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008242:Lysogeny; D011817:Rabbits; D011995:Recombination, Genetic; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics; D013604:T-Phages / Q000235:genetics*; D014158:Transcription, Genetic; D014733:Vibrio / Q000201:enzymology / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D005914:Globins; D008156:Luciferases; D006026:Glycoside Hydrolases; D001616:beta-Galactosidase; C022361:6-phospho-beta-galactosidase