Difference between revisions of "Part:BBa K4477005:Design"
(→Design Notes) |
|||
(3 intermediate revisions by the same user not shown) | |||
Line 9: | Line 9: | ||
Amino acid sequence reverse transcribed and codon optimized for expression in E. coli B (the parent strain of SHuffle). | Amino acid sequence reverse transcribed and codon optimized for expression in E. coli B (the parent strain of SHuffle). | ||
+ | His tag and TEV cleavage sites were included upstream of the antibody. The His tag ensured the entire polypeptide could be purified out of a total protein solution using immobilized metal ion affinity chromatography (IMAC), and the TEV cleavage site ensured that TEV could be used to cleave off the His tag once it was no longer needed. | ||
+ | For detailed annotations, see Team Virginia 2022's Benchling folder here: https://benchling.com/mrkhoury/f_/YR4zZSUb-virginia-igem-2022-dna-constructs/ | ||
===Source=== | ===Source=== | ||
− | https://rupress.org/jcb/article/217/3/1143/39133/A-toolbox-of-anti-mouse-and-anti-rabbit-IgG | + | 1. https://rupress.org/jcb/article/217/3/1143/39133/A-toolbox-of-anti-mouse-and-anti-rabbit-IgG |
===References=== | ===References=== | ||
+ | 2. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4983233/ |
Latest revision as of 08:12, 12 October 2022
Camelid anti-mouse VHH antibody fragment
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]
Design Notes
Amino acid sequence reverse transcribed and codon optimized for expression in E. coli B (the parent strain of SHuffle).
His tag and TEV cleavage sites were included upstream of the antibody. The His tag ensured the entire polypeptide could be purified out of a total protein solution using immobilized metal ion affinity chromatography (IMAC), and the TEV cleavage site ensured that TEV could be used to cleave off the His tag once it was no longer needed.
For detailed annotations, see Team Virginia 2022's Benchling folder here: https://benchling.com/mrkhoury/f_/YR4zZSUb-virginia-igem-2022-dna-constructs/
Source
1. https://rupress.org/jcb/article/217/3/1143/39133/A-toolbox-of-anti-mouse-and-anti-rabbit-IgG