Difference between revisions of "Part:BBa K4260111:Design"

Revision as of 21:39, 30 September 2022

Design

Fig.1:RecA intein mediated biosensor behavior in the presence endocrine disrupting chemicals (EDCs)..

RecA intein ESR1 biosensor consists of two inteins capable joining two protein fragments and separating from them. The ESR1 biosensor gene coding for the hERalpha protein was introduced with its respective linker, with the purpose of separating the N-terminal and C-terminal RecA inteins in the presence of EDCs; when the ESR1 biosensor binds to an EDC, the inteins may carry protein spicing, ligate the two N and C exteins and separate the ESR1 biosensor, ESR1 linker and the two N and C extein complex. 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 [1]. Moreover two aminoacid mutationes (Val67Leu and Asp111Glu) to make the RecA intein a stable protein [2]. The functionality of the Barcelona 2020 team's inteins from the biobrick [BBa_K348400] was verified to check the functibility of the HERα enzyme to be used in the biosensor, since it is employed as a receptor enzyme for EDCs.

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-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 [<html><a href='https://parts.igem.org/Part:BBa_K592009'>BBa_K592009</a></html>].
+RecA intein ESR1 biosensor consists of two inteins capable joining two protein fragments and separating from them. The ESR1 biosensor gene coding for the hERalpha protein was introduced with its respective linker, with the purpose of separating the N-terminal and C-terminal RecA inteins in the presence of EDCs; when the ESR1 biosensor binds to an EDC, the inteins may carry protein spicing, ligate the two N and C exteins and separate the ESR1 biosensor, ESR1 linker and the two N and C extein complex. The two endogenous fragments encode the chromoprotein AmilCP [<html><a href='https://parts.igem.org/Part:BBa_K592009'>BBa_K592009</a></html>].
 RecA intein was build from the first 110 and last 58 aminoacids from the wildtype RecA full-lenght intein [1]. Moreover two aminoacid mutationes (Val67Leu and Asp111Glu) to make the RecA intein a stable protein [2].