(bio-dsDNA) containing aT7 promotersequence, and Cas13a was used in a dual functional manner to enhance signal amplification by T7 RNApolymerasetranscription and to induce thetrans-cleavage activity. The bio-dsDNA was adopted instead of the bio-RNA because of the big size of Cas13, which ...
Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase (99 kDa) that catalyzes in vitro RNA synthesis from a DNA sequence containing a T7 phage promoter.The highly concentrated version T7 RNA Polymerase HC is ideally suited for set-up and optimization of high yield in vitro ...
Here, we use 5′RACE-Seq to screen a randomized initially transcribed region of the T7 promoter for cross-talk with transcriptional activity. We reveal that sequences from position +4 to +8 downstream of the transcription start site affect T7 promoter activity over a 5-fold range, and...
Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase (99 kDa) that catalyzes in vitro RNA synthesis from a DNA sequence containing a T7 phage promoter. The highly concentrated version T7 RNA Polymerase HC is ideally suited for set-up and optimization of high yield in vitro transc...
During initiation, the NTD of T7 RNAP binds to the promoter sequence, forming the initiation complex (IC; Fig. 1a)12,15,16. This IC is unstable and produces short RNA transcripts known as abortive transcripts of 2–10 nucleotides in length, in a process called abortive cycling (Fig. 1b...
Consensus promoter sequences for phage RNA polymerases. The promoters for RNA polymerases of T7, T3, SP6, K11, and BA14 phages share a highly conserved structure composed of 23 bp (112). Using the 23-bp consensus sequence of 17 natural T7 promoters as a reference, other consensus phage ...
Optimization of Cephalosporin C Acylase Expression in Escherichia coli by High-Throughput Screening a Constitutive Promoter Mutant library Article 06 January 2021 ReToAd: simple method for the rapid replacement of promoters to improve protein production Article 02 April 2018 Screening of an Escherich...
Depending on the RNA sequence individual optimization may increase product yield (0.5h–4h at 37°C).Component Volume Final conc. PCR-grade water Xμl HighYield T7 Reaction Buffer (10x) 2μl 1x DTT (100 mM) 2μl 10 mM ATP (100 mM) 1.5 μl 7.5 mM UTP (100 m...
subtilis phage SPO1 and the 3ʹ-sequences of the E. coli lac promoter including its operator region Fused groES promoter and lacO operon and optimization of nucleotides at the con‑ served regions of the groESL promoter (Bhavsar et al. 2001; Kim et al. 1996) (Yang et al. ...
Despite the limitations of this method, it can reveal important aspects of transcription in a rapid manner and allow optimization of features like reaction conditions and promoter sequence. It should also translate well to in vitro characterization of other polymerases, including viral, bacterial, and...