The T7 polymerase requires, at the minimum, for the promoter sequence to be double-stranded. The rest of the template does not necessarily have to be double-stranded.Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.订购...
A comparison of the sequences of other phage RNAPs and their promoters suggested additional contacts that might be important in promoter recognition. We have found that changing the amino acid residue at position 758 in T7 RNAP results in an enzyme with altered specificity for the bp at ...
Comparison of the Rescue Efficiency of Sendai Virus Minigenome Mediated by CMV and T7 Promoter 引用 收藏 分享 摘要 构建一种以分泌型荧光素酶基因(Gluc)作为报告基因的仙台病毒BB1株微小基因组质粒,比较了CMV启动子与T7启动子对仙台病毒微小基因组的拯救效率。首先设计并合成锤头状核酶序列,仙台病毒trailer、...
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 ...
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...
The raw fluorescence data for all normalized data points are on Figure S8, and the method for calculation of error bars (error propagation) is described in Materials and Methods section “Promoter comparison using different extracts”. b Apta-Nucleic Acid Sequence Based Amplification reaction ...
Embedding the aptamer in an insulator structure, such as the F30 three-way junction scaffold (50), may help avoid folding and sequence-dependent biases in a manner similar to the use of a self-cleaving ribozyme (28). Fluorescence-based mutagenesis scanning of the T7 RNAP promoter sequence ...
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...
The expression frame of the target gene, which contains all the expression elements (i.e., T7 promoter, ribosome binding site sequence (RBS), the gene of interest, termina- tor) is highly similar to the pET21a vector, except that E. coli RBS (i.e., AAGGA) was replaced with the B...
transcription reactions. The phage promoter sequences can be incorporated into the DNA by using primers that flank the phage promoter sequences in the vector or by having the promoter sequence within the 5´ oligomer used in the PCR. The resulting PCR-generated DNA can be purified using the...