What is T7 and SP6?

What is T7 and SP6?

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Q. What is T7 and SP6?

SP6 RNA Polymerase is used for the synthesis of RNA transcripts in the 5´→ 3´ direction from vectors containing the SP6 phage promoter, while T7 RNA Polymerase catalyzes the synthesis of RNA in the presence of a DNA template containing T7 phage promoter.

Q. What is the SP6 promoter?

The SP6 promoter sequence is 5´ ATTTAGGTGACACTATAG 3´. SP6 RNA Polymerase starts transcription at the underlined G in the double-stranded promoter sequence. The polymerase then transcribes using the opposite strand as a template in the 5´ to 3´ direction.

Q. Is T7 double-stranded?

T7 RNA polymerase requires a double-stranded DNA promoter in order to initiate transcription; however, elongation does not require a double-stranded DNA template. This method allows rapid production of a variety of tRNA transcripts which can be aminoacylated well.

Q. How does T7 work?

T7 RNA polymerase is a very active enzyme: it synthesizes RNA at a rate several times that of E. coli RNA polymerase and it terminates transcription less frequently; in fact, its transcription can circumnavigate a plasmid, resulting in RNA several times the plasmid length in size.

Q. What is T7 tag?

The T7 tag is an 11 amino acid peptide encoded in the leader sequence of T7 bacteriophage gene10. This gene encodes a T7 major capsid protein whose function is not clear. Monoclonal antibodies specific for T7 tag are an important tool for studying expression of recombinant T7-tagged proteins.

Q. How does the T7 promoter work?

Q. What makes a strong promoter?

The specific sequence of the promoter determines the strength of the promoter (a strong promoter leads to a high rate of transcription initiation). The presence or absence of the protein will affect the strength of the promoter.

Q. Is the T7 promoter inducible?

The second contains the T7 RNA polymerase gene under the control of a heat-inducible E. coli promoter. Upon heat induction, the T7 RNA polymerase is produced and initiates transcription on the expression vector, resulting in turn in the expression of the gene(s) under the control of p(T7).

Q. Is there vaccine for T7?

T7 phage may be potentially useful as a delivery vector for DNA vaccine transfer. The surface display capability of T7 phage also enlarge the use in vaccine design, for it can surface display antigen epitope and carry DNA vaccine within one particles.

Q. What are the binding sites for Sp6 and T7?

As Bob1 said, these are polymerase binding sites typically used for in vitro transcription/translation reactions where the Sp6 or T7 polymerase is added to the mix in a rabbit reticulocyte or wheat germ background.

Q. Why do you need a T7 and SP6 promoter?

They are actually RNA polymerase binding sites designed put there to allow expression of the cloned gene. Not just there for primer binding. However, if you rely on them, you’ll need to have a host strain that expresses either the T7 or the Sp6 RNA polymerase, depending on which promoter you want to use.

Q. Where do T7 and SP6 RNAPs terminate?

This means that T7 and SP6 RNAPs proceed to the end of the DNA template, and do not terminate at any specific sites within the plasmid. For this reason, the circular plasmid template should be linearized by restriction digestion prior to in vitro transcription.

Q. How does the T7 vertebra protect the spinal cord?

The T7 vertebra also plays an important role in the protection of the spinal cord. The spinal cord runs through the vertebral foramen and is protected by the centrum and the vertebral arch that surround it.

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