Random insertion of a TetR-inducing peptide tag into Escherichia coli proteins allows analysis of protein levels by induction of reporter gene expression.
The insertion element InsTipalpha was constructed to generate protein expression data. It randomly fuses the TetR-inducing peptide Tip to the affected reading frame. Fusion protein expression is quantified by Tet-regulated reporter gene expression. The expression patterns of tagged Escherichia coli genes fully agree with published data from transcriptional fusions or microarrays, validating the Tip tag approach.
Schlicht
Maximilian
M
Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstrasse 5, D-91058 Erlangen, Germany.
Berens
Christian
C
Daam
Janko
J
Hillen
Wolfgang
W
D004251:DNA Transposable Elements / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D009154:Mutation; D010455:Peptides / Q000235:genetics* / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013753:Tetracycline Resistance / Q000235:genetics; D014158:Transcription, Genetic
D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; C470187:TetR(B) protein, E coli