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Record number: 1, Textual support number: 1 CONT

Purification of mutant enzymes is a prime requirement of biophysical and biochemical studies. Our investigations on the essential E. Coli enzyme glutaminyl-tRNA sybthetase demand mutant enzymes free of any wild-type protein contamination. However, as it is not possible to express non-complementing mutant enzymes in an E. Coli gInS-deletion strain, we developed a novel strategy to address these problems. Instead of following the common tactic of epitope-tagging the mutant protein of interest on an extrachromosomal genetic element, we fused a reporter epitope to the 5' end of the chromosomal gInS-gene copy: this is referred as reverse epitope-tagging.  1, fiche 1, Anglais, - reverse%20epitope%2Dtagging

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