Difference between revisions of "Part:BBa K3771035"

Revision as of 20:09, 17 October 2021

PpspA-CoaBC-6xHis-PompA-OmpA/OprF

Description

This composite part is a component of the IFN-γ sensing system and was used to express the taurine production enzyme, CoaBC.

Biology

The ompA promoter facilitates the constitutive expression of OmpA/OprF. Binding of IFN-γ to the OmpA/OprF chimeric protein induces the response of the phage shock protein (Psp) system, a highly conserved stress response system in enterobacteria[1]. Signal transduction from the outer membrane to the inner membrane activates the pspA promoter, initiating expression of CoaBC. CoaBC converts L-cystate into taurine in the taurine synthesis L-cystate pathway.

Usage

We ligased the NPO-OmpA* fragment and pspA-CoaBC-his-tag on the pSU expression vector and transformed it into DH5α to complete construction of the plasmid. The his-tag allows for confirmation of CoaBC expression by western blot using the anti-6X his-tag antibody.

Characterization

Double digestion results are shown in Fig. 2.

圖片描述

References

1. Darwin AJ. The phage-shock-protein response. Molecular Microbiology. 2005;57(3):621-628. doi:10.1111/j.1365-2958.2005.04694.x https://pubmed.ncbi.nlm.nih.gov/16045608/
Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 12
Illegal BamHI site found at 2533
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 727
1000
COMPATIBLE WITH RFC[1000]

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 ===Usage and Biology===