T7 RNA polymerase variants with Cysteine-Serine substitutions

摘要

The present invention aims at providing the skilled person with novel variants of T7 polymerase. A T7 variant according to the invention includes a Cysteine-Serine substitution on position 723 of the amino acid sequence of the T7 polypeptide. A T7 variant according to the invention has a DNA-dependent RNA polymerase enzymatic activity and a reduced tendency to form intramolecular homodimers by way of oxidizing thiol groups. The identified amino acid substitutions according to the invention appear to impact only minimally, if at all, on RNA polymerase activity of the T7 polypeptide. The mutations of the invention can optionally be combined with even further mutations, including mutations which provide enhanced thermostability compared to the wild-type reference.