Part:BBa_K1162009
10x-Histidine (10x-His) Tag with Met (ATG) For N-terminal purification
Design
This part was designed with Assembly Standard #23 (Silver Fusion) to allow for protein fusions. This 10x His-Tag is an improvement upon BBa_K844000 by removing the double stop codon (TAA TAA) to allow for C-terminal fusion of proteins. Another key characteristic of this part is the presence of a Methionine(atg) at the start of the coding region (N-terminal), which enables the user to fuse proteins downstream and then purify. The promoter-rbs system of choice can be placed in front of this His-Tag for protein expression.
Experience
Figure 1. The above image shows a GFP dot image (on parafilm, illuminated by UV light) in which it is clear that the N-terminal 10x His-Tag (BBa_K1162009) method of purification works as desired and adds another important purification system to the registry.
Figure 2. From the SDS-PAGE gel it can be seen that there is pure GFP (~26.9 kDa) in elution fractions number 2 and 3. The wash fractions do not appear to have any GFP, which indicates that the N-terminal 10x His-Tag is strongly bound to the Nickel resin column during washing steps.
Related parts
BBa_K844000 - this link also provides an extensive review of existing Histidine Tags in the Parts Registry.
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]
| n/a | 10x-Histidine (10x-His) Tag with Met (ATG) For N-terminal purification |