Difference between revisions of "Part:BBa K2872003:Design"
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===Biology and Usage=== | ===Biology and Usage=== | ||
− | This part codes for human influenza hemagglutinin (HA) molecule and is fused with a stop codon (taa) at the C-terminus. | + | This part codes for 3x human influenza hemagglutinin (HA) molecule and is fused with a stop codon (taa) at the C-terminus. |
− | + | '''HA tag:''' | |
An HA-tag is derived from the human influenza hemagglutinin (HA) molecule corresponding to amino acids 98-106. It has been extensively used as a general epitope tag in expression vectors. The HA-tag does not appear to interfere with the bioactivity of the recombinant HA-tagged protein. [1] | An HA-tag is derived from the human influenza hemagglutinin (HA) molecule corresponding to amino acids 98-106. It has been extensively used as a general epitope tag in expression vectors. The HA-tag does not appear to interfere with the bioactivity of the recombinant HA-tagged protein. [1] | ||
+ | <br> | ||
− | + | '''Purification:''' | |
The HA-tagged protein binds to the HA-tag specific monoclonal antibody conjugated on an agarose gel. After washing away residual impurities, bound HA-tag proteins can be eluted off the affinity column by high concentration of the HA-tag peptide or by low pH buffer. An EK cleavage site behind the HA-tag (HA-EK site-protein structure) can allow complete removal of the HA-tag and the cleavage site, leaving no additional amino acids after the specific cleavage of the HA-tag. [2] | The HA-tagged protein binds to the HA-tag specific monoclonal antibody conjugated on an agarose gel. After washing away residual impurities, bound HA-tag proteins can be eluted off the affinity column by high concentration of the HA-tag peptide or by low pH buffer. An EK cleavage site behind the HA-tag (HA-EK site-protein structure) can allow complete removal of the HA-tag and the cleavage site, leaving no additional amino acids after the specific cleavage of the HA-tag. [2] | ||
Latest revision as of 11:24, 10 October 2018
Human influenza hemagglutinin (HA) tag
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 641
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 628
Biology and Usage
This part codes for 3x human influenza hemagglutinin (HA) molecule and is fused with a stop codon (taa) at the C-terminus.
HA tag:
An HA-tag is derived from the human influenza hemagglutinin (HA) molecule corresponding to amino acids 98-106. It has been extensively used as a general epitope tag in expression vectors. The HA-tag does not appear to interfere with the bioactivity of the recombinant HA-tagged protein. [1]
Purification: The HA-tagged protein binds to the HA-tag specific monoclonal antibody conjugated on an agarose gel. After washing away residual impurities, bound HA-tag proteins can be eluted off the affinity column by high concentration of the HA-tag peptide or by low pH buffer. An EK cleavage site behind the HA-tag (HA-EK site-protein structure) can allow complete removal of the HA-tag and the cleavage site, leaving no additional amino acids after the specific cleavage of the HA-tag. [2]
Source
Human influenza hemagglutinin is found on the surface of influenza viruses.
References
[1] “Anti-HA Tag.” Razor Tie Artery Foundation Announce New Joint Venture Recordings | Razor & Tie, Rovi Corporation, web.archive.org/web/20100914142422/http://www.millipore.com/catalogue/item/05-904.
[2] Schembri, Laura, et al. “The HA Tag Is Cleaved and Loses Immunoreactivity during Apoptosis.” Nature Methods, vol. 4, no. 2, 2007, pp. 107–108., doi:10.1038/nmeth0207-107.