Part:BBa_K5160012
HA
Overview
The HA tag (HA-tag) is a protein tag derived from the human influenza hemagglutinin (HA) antigen, with its chemical essence being a short amino acid sequence spanning the 98-106th amino acids of the human influenza HA molecule, specifically YPYDVPDYA. The HA tag is extensively used as an epitope tag in expression vectors, facilitating protein detection, isolation, and purification, as it does not disrupt the protein's biological activity or distribution. By appending the HA tag to the target protein, specific antibodies against the HA tag can bind to the recombinant protein, enabling convenient experiments such as immunohistochemistry (IHC) and Western Blotting. HA antibodies can also be immobilized onto agarose beads to enhance protein purification efficiency. Numerous recombinant proteins can express the HA tag, aiding in the rapid acquisition of information regarding their localization, expression levels, or biological functions.
In our project design, to purify and identify the target protein while amplifying the detection signal and boosting purification efficiency, we have fused the HA tag protein to the C-terminus of the target protein to create the corresponding vector. Subsequently, we introduce the target gene into the tomato genome sequence through transient infection technology or transgenic techniques. When the target gene is expressed, the HA tag protein is also expressed, which can be utilized for subsequent Western blot (WB) detection and co-immunoprecipitation to confirm the expression of the target protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design
1. During the design process, considering the need to add the vacuolar signal peptide SPS-NTPP to the N-terminus of the target sweet protein, we deliberately placed the HA tag at the C-terminus of the target protein to prevent it from interfering with the SPS-NTPP vacuolar signal peptide.
2.Considering that the HA tag can be cleaved by Caspases 3 and 7, resulting in the loss of immunoreactivity, we have refrained from using the HA tag for apoptotic cell proteins. Additionally, we have strictly controlled the storage conditions of the HA tag peptide to ensure its stability and activity.
Source
The HA tag originates from the 98-106th amino acid residues of the human influenza virus HA molecule and is a polypeptide with a length of 9 amino acids. The molecular weight of the HA tag is relatively small, exerting minimal influence on the spatial structure of the target protein and not interfering with the protein's biological activity or biodistribution.
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