Difference between revisions of "Part:BBa K4260111"
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Revision as of 06:26, 30 September 2022
RecA mini intein mediated ESR1 protein with OmpA signal peptide, linker and AmilCP.
This part encodes an intein mediated biosensor, consisting of a divided AmilCP chromoprotein gene, a mutated intein RecA N-terminal and C-terminal domains and a ESR1 Ligand Binding Domain. The RecA intein comes from Mycobacterium Tuberculosis (Mtu) which has been mutated with the purpose of making it more stable.
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
This RecA intein ESR1 biosensor consists of two inteins capable of joining and separating from two protein fragments. Considering this, the ESR1 biosensor gene coding for the hERalpha protein was introduced with its respective linker, in such a way that when an endocrine disruptor binds to the biosensor, the inteins join and separate with the rest of the protein, removing the linker, the biosensor and the two inteins. The two endogenous fragments encode the chromoprotein AmilCP [BBa_K592009].
RecA intein was build from the first 110 and last 58 aminoacids from the wildtype RecA full-lenght intein. Moreover two aminoacid mutationes (Val67Leu and Asp111Glu) to make the RecA intein a stable protein.