Part:BBa_K4488014
Fusion of free-use GFP with CBDcenA (cellulose-binding domain) at the C-terminal end with a linker
The construct can be cloned into an expression vector such as pET28c in E.coli to produce a fusion protein of fuGFP with CBDcenA. The fuGFP sequence is towards the N terminus of the protein with CBDcenA (BBa_K1321339) downstream followed by a stop codon. Recognition sites for BamHI and BsaI are present before the RBS allowing golden gate cloning. XhoI and BsaI are also present downstream of the stop codon.
Usage and Biology
Our project provided preliminary evidence that CBD and cellulose can be used to purify proteins by using fuGFP-linker-CBDcenA.
The linker improved the solubility and fluorescence of the sequence. Below is the fluorescence assay depicting a higher reading compared to our previous construct BBa_K4488010:
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 163
- 1000COMPATIBLE WITH RFC[1000]
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