Part:BBa_K1893018
Arabinose inducible 3-isopropylmalate dehydrogenase (pBAD+leuB)
This composite part consists of a leuB coding sequence under the control of an arabinose-inducible promoter (BBa_K1893015). leuB encodes for 3-isopropylmalate dehydrogenase, an enzyme in the leucine biosynthesis pathway that catalyses the oxidation of 3-isopropylmalate to 3-carboxy-4-methyl-2-oxopentanoate.
Usage and Biology
We initially considered auxotrophy as a method of controlling growth through our growth-regulation module. The pBAD+leuB was designed to investigate the effect of leuB suppression on the growth of E. coli leuB knockout strains. The knockout strains were to be grown in media lacking leucine, so that only cells expressing leuB would be capable of growth. Once the effect of leuB expression on growth in starvation conditions had been determined, we would then place leuB under the control of an inverter. As a result, our STAR part could activate expression of the inverter, which would suppress leuB expression and lead to growth inhibition.
However, introducing an inverter into the system would have greatly increased the complexity of our circuit. Furthermore, we realized that having to generate leuB knockout strains and media lacking leucine contradicted the design specifications of our system, which aimed to make the implementation of co-culture experiments as simple as possible. As a result, we decided against further pursuing leuB as a method of growth control.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1342
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1281
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1116
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1098
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