promoter strengthtranscription processivitytRNA reporter geneIn vitro , after binding to the promoter to form a catalytically active complex, RNA polymerases abortively cycle over the first transcribed nucleotides (initial transcribed sequence or ITS) before leaving the promoter. With the bacteriophage T7 ...
RT extends the primer into a complementaryDNA sequence. The RNA strand of the formed RNA/DNA duplex is then degraded byRNase Hthus allowing the reverse primer P2 to hybridize the ssDNA and RT to extend P2 primer thus producing a dsDNA that contains the T7 promoter sequence. After the initia...
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 ...
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...
vitro transcription/translation rom the T7 promoter, located between the GAL4 AD and HA tag sequences, produces an HA-tagged protein that lacks the GAL4 AD. Such proteins can be detected by the HA-Tag Polyclonal Antibody, but not the GAL4 AD Monoclonal Antibody. ...
It also provides a binding site for sequencing with the T7 Promoter Sequencing Primer. pGADT7-Rec2 is a shuttle vector; it can be maintained in both yeast and bacteria. It contains an autonomous replication sequence (ARS4) and a LEU2 nutritional marker for replication and selection in yeast...
It will transcribe large amounts of RNA from DNA sequences [such as, plasmids, polymerase chain reaction (PCR) fragments, or hybridized oligonucleotides] downstream of its promoter, without cross-talk from nearby SP6 or T3 promoters. Thus, RNA molecules transcribed from a linear template will be...
• T7 RNA polymerase promoter: 1905–1927 • HA epitope tag: 1942–1968 • SMART III Oligonucleotide sequence: 2001–2036 • CDS III Primer sequence: 2047–2071 • Transcription termination signal Fragment carrying the S. cerevisiae ADH1 terminator (T ...
The coding sequence for the desired RNA or protein (referred to as the target RNA or protein) is typically placed in a plasmid under control of a T7 promoter, that is, a promoter recognized specifically by T7 RNA polymerase. In the absence of an inducer for the lacUV5 promoter, little ...
1. A DNA sequence comprising a regulatable promoter operatively linked to a gene encoding a truncated bacteriophage T7 RNA polymerase capable of binding to a T7 promoter and carrying out transcription, which gene comprises a nucleotide sequence corresponding to the sequence of FIG. 16, wherein one...