Difference between revisions of "Part:BBa K1362453"
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This part codes for the amino acid sequence ENLYFQG. This site is recognized by TEV protease (catalytic domain of the Tobacco Etch Virus nuclear inclusion a (NIa) protein), which will cleave the Gln-Gly peptide bond [[{{PAGENAME}}:Design#References|[1]]]. The final four nucleotides of this sequence are GGGT, which will be the overhang produced wenn a reverse-facing BsaI site (<partinfo>BBa_K1362423</partinfo>, <partinfo>BBa_K1362427</partinfo>, <partinfo>BBa_K1362447</partinfo>) is directly following this part, as in the Sortase A circularization constructs (<partinfo>BBa_K1362202</partinfo>, <partinfo>BBa_K1362203</partinfo>, <partinfo>BBa_K1362204</partinfo>, <partinfo>BBa_K1362205</partinfo>). | This part codes for the amino acid sequence ENLYFQG. This site is recognized by TEV protease (catalytic domain of the Tobacco Etch Virus nuclear inclusion a (NIa) protein), which will cleave the Gln-Gly peptide bond [[{{PAGENAME}}:Design#References|[1]]]. The final four nucleotides of this sequence are GGGT, which will be the overhang produced wenn a reverse-facing BsaI site (<partinfo>BBa_K1362423</partinfo>, <partinfo>BBa_K1362427</partinfo>, <partinfo>BBa_K1362447</partinfo>) is directly following this part, as in the Sortase A circularization constructs (<partinfo>BBa_K1362202</partinfo>, <partinfo>BBa_K1362203</partinfo>, <partinfo>BBa_K1362204</partinfo>, <partinfo>BBa_K1362205</partinfo>). | ||
| + | <ul> | ||
| + | <li>TEV Protease is a recombinant protease derived from Tobacco Etch Virus (TEV) Nla, which is used to excise the affinity tag of the purified fusion protein. TEV protease has strong site specificity and can recognize the seven amino acid sequence of EXXYXQ(G/S) (Glu-Asn-Leu- Tyr-Phe-Gln-Gly). The most common one is ENLYFQG, and its cleavage site is at Between glutamine and glycine or serine.</li> | ||
| + | </ul> | ||
| + | ==Experimental Results== | ||
| + | [[File:T--LZU-CHINA--TEVp.png|600px|thumb|center|]] | ||
| + | <ul> | ||
| + | <li>Using the specificity of TEVp, we cleave the recognition sites of ALKBH (Fig.A) and Casp3 (Fig.B) linked by the target sequence, which are active.</li> | ||
| + | </ul> | ||
| + | ===Usage and Biology=== | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 09:19, 21 October 2019
TEV protease recognition/cleavage site
This part codes for the amino acid sequence ENLYFQG. This site is recognized by TEV protease (catalytic domain of the Tobacco Etch Virus nuclear inclusion a (NIa) protein), which will cleave the Gln-Gly peptide bond [1]. The final four nucleotides of this sequence are GGGT, which will be the overhang produced wenn a reverse-facing BsaI site (BBa_K1362423, BBa_K1362427, BBa_K1362447) is directly following this part, as in the Sortase A circularization constructs (BBa_K1362202, BBa_K1362203, BBa_K1362204, BBa_K1362205).
- TEV Protease is a recombinant protease derived from Tobacco Etch Virus (TEV) Nla, which is used to excise the affinity tag of the purified fusion protein. TEV protease has strong site specificity and can recognize the seven amino acid sequence of EXXYXQ(G/S) (Glu-Asn-Leu- Tyr-Phe-Gln-Gly). The most common one is ENLYFQG, and its cleavage site is at Between glutamine and glycine or serine.
Experimental Results
- Using the specificity of TEVp, we cleave the recognition sites of ALKBH (Fig.A) and Casp3 (Fig.B) linked by the target sequence, which are active.
Usage and Biology
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]