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Parts

Introduction

The majority of parts in the iGEM registry are designed to work in E. coli, however our project is centred around genetic modification of G. hansenii. In particular, the biobricks to produce the light sensitive signal transduction pathway based around Cph8(see Project Background) do not include a gene for the expression of OmpR because it is found natively in E. coli and is therefore not required for functional expression when working in E. coli.

Aims

• Produce a biobrick containing ompR and sequence it to provide a reliable part for future use in expression of the the Cph8 light sensitive signal transduction pathway in non E. coli chassis.

• Provide clarifying notes in the registry on parts which have confusingly used the term "ompR" or "OmpR" to label the ompC promoter (ompCp) sequence. Or in some cases have labelled the sequence of ompR as "envZ".

• Produce a composite biobrick for use in a broad host-range vector such as pSEVA321, pSEVA331, pSEVA351 which will show whether a a given non-E. coli chassis can functionally express the Cph8 light sensitive signal transduction pathway.

• Produce a composite biobrick to control the production of extracellular cellulose in dgc1 knockout mutants of G. hansenii.

Parts

BBa_K1337000
ompR from E.coli

BBa_K1337001
dgc1 from G. hansenii

BBa_K1337002
Cph8 signal transduction pathway compatibility tester.

BBa_K1337003
Light mediated dgc1 transcription construct (should produce the cellular pathway in Figure 1.)

Results

So far we have successfully amplified envZ from E. coli (Figure 2 - gel electrophoresis of PCR product from primers flanking envZ from E. coli).

As we continue the project, we hope to successfully amplify ompR from E. coli and ligate it into pSB1C3, in order to submit our first independently produced biobrick.

Following on from that, we hope to also produce and submit our other planned plasmids.