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. 1993 Apr 15;90(8):3147-51.
doi: 10.1073/pnas.90.8.3147.

T7 RNA polymerase mutants with altered promoter specificities

C A Raskin  1 , G A Diaz, W T McAllister
Affiliations

T7 RNA polymerase mutants with altered promoter specificities

C A Raskin et al. Proc Natl Acad Sci U S A. .

Abstract

The amino acid at position 748 in T7 RNA polymerase (RNAP) functions to discriminate base pairs at positions -10 and -11 in the promoter. We have constructed a series of T7 RNAP mutants having all possible amino acid substitutions at this position. Surprisingly, most (13/19) substitutions result in active RNAPs, and many of these exhibit altered promoter specificities. Identification of mutant RNAPs with altered specificities expands the repertoire of highly specific phage RNAPs that are available for use in phage RNAP-based transcription systems and highlights the complexity of sequence-specific DNA recognition.

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