Difference between revisions of "Part:BBa K629012"

Latest revision as of 09:19, 10 October 2011

T7 promoter + lac operator + RBS +cheZ

Used to test the chemotaxis when expression is controlled by a strong promoter.

Background

Polar localization is dependent on CheA-short. CheZ is a cytosolic phosphatase which functions in the chemotaxis signal transduction complex. Plays an important role in bacterial chemotaxis signal transduction pathway by accelerating the dephosphorylation of phosphorylated CheY (CheY-P).

Expression in E. Coli

Before we started our functional test of CheZ, we have utilized western blot to determine whether there are enough soluble proteins in E. Coli. With 6×His tag on pET-32a, the result shows that CheZ has more expression with induction of IPTG.

Note: Because the promoter is Plac here, the expression has a background, which means control also expresses some cheZ without induction of IPTG.

Picture of Chemotaxis in cheZ knocked-out strain

The main purpose of cheZ is to control chemotaxis in E. Coli. What we have done is to use different promoter to test how to get the better result from expression of CheZ. Below are pictures and parameters of chemotaxis test.

Nalidixic acid Induction of promoter recAp. If you need higher resolution pictures, please feel free to cantact us.

Acknowledgement

Thanks Dr. Parkinson very much for providing cheZ knocked-out strain.

Sequence and Features