Part:BBa_S05054:Experience

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Applications of BBa_S05054

When we finished the combination of BBa_K081024 and BBa_I13507(mRFP), we were astonished to find out mRFP was expressed without Cph8. Meanwhile, the sequence is proved to be correct.

After blasting the sequence of Cph1 at NIBC (http://www.ncbi.nlm.nih.gov), we discovered that the BL21(DE3) strain has endogenous EnvZ, which is the key part of Cph1that can phosphorylate the OmpR and cause the activation of OmpC followed by the expression of downstream genes, BBa_I13507. Hence, once the strain has endogenous EnvZ, BBa_K081024 & BBa_I13507, also known as BBa_S05054 expresses mRFP without Cph8 not to mention the regulation of red light.

Now it is confirmed that as long as equipped with a promoter,RBS and BBa_I15008, the OmpC promoter in BBa_S05054 can work normally in a endogenous-EnvZ-free system. Besides, BBa_S05054 can also be utilized to test the exisitence of endogenous EnvZ.

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