Difference between revisions of "Part:BBa K4260001:Design"
| Line 25: | Line 25: | ||
|} | |} | ||
| − | Design | + | === Design === |
The TecCEM team 2022 designed this sequence for the codification of the Human Estrogen Receptor Alpha (hERa), this is a receptor protein which aim is to bind to estrogens. This protein keeps important amino acids sites Gly521, His524, Leu525 and Met528. It is also used as the biological receptor of some endocrine disrupting chemicals. | The TecCEM team 2022 designed this sequence for the codification of the Human Estrogen Receptor Alpha (hERa), this is a receptor protein which aim is to bind to estrogens. This protein keeps important amino acids sites Gly521, His524, Leu525 and Met528. It is also used as the biological receptor of some endocrine disrupting chemicals. | ||
| Line 33: | Line 33: | ||
Figure 1. Construct sequence design. | Figure 1. Construct sequence design. | ||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
Revision as of 14:33, 24 September 2022
ESR1: Estrogen Receptor 1 with periplasmic signal peptide OmpA, GGGGSC linker and histidine tag
| ESR1_HD shortlist resume | |
|---|---|
| Function | Periplasmic expression of Human
Estrogen Receptor Alpha protein |
| Optimization for | E.coli strains |
| Signal Peptide | OmpA-periplasmic expression |
| Linker | GGGGSC - create disulfide bonds
with chitosan |
| Added tags | Histidine tag for Nickel Column |
| Submitted by | [Link to wiki] |
Design
The TecCEM team 2022 designed this sequence for the codification of the Human Estrogen Receptor Alpha (hERa), this is a receptor protein which aim is to bind to estrogens. This protein keeps important amino acids sites Gly521, His524, Leu525 and Met528. It is also used as the biological receptor of some endocrine disrupting chemicals.
Therefore, we used the genomic coding sequence of Homo sapiens Estrogen Receptor 1 (ESR1) optimizing its codons for an E.coli expression. We added a (GGGGSC) linker, this one is composed of four glycines, one serine and one cysteine, with the purpose of attaching hER alpha protein to chitosan, thus, ensuring the desired position of the molecule showing the estrogen binding site up for an effective capture. The periplasmic signal peptide element “OmpA” helps the cellular machinery to speed up the process of protein expression and send it to the periplasmic space, where it can be purified using the histidine tag for a nickel column. Figure 1 illustrates the detailed design of this part.
Figure 1. Construct sequence design.