Difference between revisions of "Part:BBa K3771015"

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		+	<p align="center">Fig. 2. Confirmation of protein expression by western blot. Lane 1: OmpA positive control (~35 kDa); Lane 2: negative control; Lane 3, 4: OmpA/OprF chimeric protein (~39kDa)</p>
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Latest revision as of 19:43, 19 October 2021

PlacI-OmpA/OprF

Description

P_lacI-ompA/oprF is part of the IFN-γ sensing system. lacI promoter regulates the constitutive expression of OmpA/OprF.

Biology

When there is no lac repressor, the lacI promoter facilitates the constitutive expression of OmpA/OprF. In the IFN-γ sensing system, binding of IFN-γ to the OmpA/OprF chimeric protein activates the pspA promoter, producing the enzyme required for the synthesis of taurine.

Usage

We ligased ompA/oprF gene and the P_lacI expression vector pSU and transformed it into DH5α to complete construction of the plasmid. Then, the plasmid was purified and transformed into E. coli ompA mutant strain JW0940.

Characterization

Fig. 1. PCR confirmation of P_lacI-ompA/oprF

Western blot was performed to confirm the expression of OmpA/OprF

Fig. 2. Confirmation of protein expression by western blot. Lane 1: OmpA positive control (~35 kDa); Lane 2: negative control; Lane 3, 4: OmpA/OprF chimeric protein (~39kDa)

Sequence and Features