T7 RNA Polymerase

Bacteriophage T7 RNA Polymerase (RNAP) is a highly processive DNA-dependent RNA polymerase that catalyzes the transcription of T7 genes1. It acts by unwinding the double-stranded DNA to provide access to the coding strand for transcription initiation (Figure 1). The 99 kDa enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under T7 promoters. RNA produced using the T7 RNAP is suitable for many applications in research and biotechnology. Intact Genomics IG® T7 RNAP is a purified recombinant T7 bacteriophage enzyme that is expressed and purified from E. coli. This enzyme is designed to perform in vitro transcription of a gene under a phage T7 promoter. The physical purity of this enzyme is ≥85% as assessed by SDS-PAGE with Coomassie® blue staining and gel densitometry (Figure 2). T7 RNA polymerase picture Fig. 1: The T7 RNAP enzyme is a protein that binds to and transcribes RNA from double-stranded DNA. A molecular surface representation of T7 RNAP in complex with DNA (PDB ID# 3e2e) is shown in the early stages of transcription. Protein residues-green, DNA-orange and blue strands, Initiating RNA strand-black. Applications Radiolabeled RNA probe preparation RNA synthesis from DNA template sgRNA and mRNA synthesis Applications in mRNA therapeutics Product Source E. coli BL21 (DE3) strain expressing a recombinant T7 RNA polymerase gene (T7 gene 1) with an N-terminal 6xHis tag. Components and Storage The T7 RNA Polymerase product contains the items below. Store all components at -20°C. T7 RNA Polymerase 25000 Units (50,000 Units/mL) Storage Buffer (50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 ºC)