Difference between revisions of "Part:BBa K3802000"
(→Sequence and Features) |
|||
| Line 2: | Line 2: | ||
===Sequence and Features=== | ===Sequence and Features=== | ||
| + | |||
| + | This fusion protein aims to anchor MlrA, or microcystinase, an enzyme capable of degrading freshwater toxin microcystin-LR (MC-LR) to the outer membrane of E. coli. This part puts together a codon-optimized subunit of PgsA (poly-γ-glutamic acid synthetase) [https://parts.igem.org/Part:BBa_K2963020 (BBa_K2963020)], an anchoring motif to MlrA with 6x His [https://parts.igem.org/Part:BBa_K1907002 (BBa_K1907002)], with a 2 amino acid Gly-Ser linker [https://parts.igem.org/Part:BBa_J18920 (BBa_J18920)]. The addition of the His tag to MlrA gives this part the ability to be purified using Nickel affinity chromatography and detected using Western blotting with anti-His antibodies. | ||
| + | This BioBrick was put under the control of an IPTG inducible ptac promoter [https://parts.igem.org/Part:BBa_K3802001 (BBa_K3802001)], as using a constitutive promoter could potentially lead to the depletion of resources for the bacteria. | ||
Revision as of 16:09, 21 October 2021
PgsA-MlrA fusion protein
Sequence and Features
This fusion protein aims to anchor MlrA, or microcystinase, an enzyme capable of degrading freshwater toxin microcystin-LR (MC-LR) to the outer membrane of E. coli. This part puts together a codon-optimized subunit of PgsA (poly-γ-glutamic acid synthetase) (BBa_K2963020), an anchoring motif to MlrA with 6x His (BBa_K1907002), with a 2 amino acid Gly-Ser linker (BBa_J18920). The addition of the His tag to MlrA gives this part the ability to be purified using Nickel affinity chromatography and detected using Western blotting with anti-His antibodies. This BioBrick was put under the control of an IPTG inducible ptac promoter (BBa_K3802001), as using a constitutive promoter could potentially lead to the depletion of resources for the bacteria.