Part:BBa_K4205000
Lpp-OmpA-MT1A
We found metallothionein (MT), a common protein utilized to remove metal ions, by consulting the literature. MT is a family of low molecular weight proteins that have a single peptide chain containing 61-68 amino acids, of which 20 are cysteines, distributed in two domains α and β clusters. Considering the practical application in human intestine,we chosed the human metallothionein MT1A to realize the function. We realized MT1A expression on membrane through Lpp-OmpA hybrid display system that no one else has ever done before. As for the sensor,we chosed the MerR family TFs. They are a large class of bacterial proteins with unique physiological functions and molecular actions: under normal conditions, they function as transcription repressors, but rapidly transform into transcription activators in response to an imbalance of cellular metal ions. In addition, for biosafety, we designed a killing switch which is inducted by an inexpensive and non-toxic monosaccharide L-arabinose. Promoter PBAD is frequently used for killing switch due to s moderately high expression levels and tight regulation of transcription. MazF is an endoribonuclease that cleaves RNAs at ACA sites and causes the death of microbe, mediates killing of cells without causing lysis of bacterial cells. We inserted the toxic protein MazF in downstream of the PBAD promoter. Therefore, upon achieving the expected effect, the expression of MazF will lead to the host bacteria lysis.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 459
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