Part:BBa_K5036003

TCS (Q, G)

Part Description

This is the TEV Cleavage Site which is a particular amino acid sequence that the TEV protease detects and cleaves

Usage

This specific amino acid sequence is recognized by TEV, which can then cleave it to liberate dcas9 from our dCas9-synRTK receptor when VEGF binds to our receptor and activates it

this figure illustrates the structure of TEV cleavage site .

literature characterization

This research investigated how adding a TEV cleavage site impacted the production and solubility of nine specific mammalian proteins within bacterial cells. It's important to note that only these nine proteins were examined, and they are not representative of a broader selection. Four distinct tags were employed in the experiment: His-L, His-S, TEV1, and TEV2

This section details the amino acid sequences of four tags attached to the N-terminus of proteins. Each tag contains six histidine residues, highlighted for easy identification. Additionally, the TEV protease cleavage site (ENLYFQ/G) is bolded for the TEV1 and TEV2 tags. It's important to note that the His-S and TEV2 tags have an extra four amino acids (SAND) at the C-terminus, compared to the other two tags.

This section examines the analysis of expression and solubility of proteins with four different His-tags. The boxes indicate the expected size of the protein produced. (A) The mouse protein USF1 was produced at high levels from E. coli when the His-L, His-S, and TEV1 tags were present, but no protein was produced when the TEV2 tag was used. The protein remained dissolved when the His-L and His-S tags were used, but the protein formed clumps when the TEV1 tag was used. (B) The mouse latexin protein was produced at high levels with all four tags, but the protein only remained dissolved when the His-L or His-S tag was present. The presence of the TEV cleavage site (in TEV1 and TEV2 tags) resulted in insoluble proteins.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]