The present invention relates to the technical field of
nucleic acid tool enzymes, and particularly to a T7
RNA polymerase mutant and its preparation and application. The T7
RNA polymerase mutant of the present invention has
RNA polymerase activity based on the T7
promoter. Compared with the wild-type T7
RNA polymerase, it has the advantages of strong
thermal stability, high transcriptional
enzyme activity, and high transcriptional product expression rate. It can also use modified nucleotides such as (m<supgt;1< / supgt;φTP) instead of
uridine triphosphate (UTP) as a substrate through
in vitro co-transcription to efficiently synthesize mRNA with high expression efficiency. Compared with the existing T7
RNA polymerase, its performance is significantly improved, better meeting the requirements of current large-scale mRNA production, and providing an effective candidate
enzyme tool for the research and application of RNA. Moreover, the method for preserving the T7
RNA polymerase mutant provided by the present invention can enable the lyophilized
powder of the T7 RNA polymerase mutant to retain more than 90% of its
enzyme activity after being stored at 4°C for 1 year.