Difference between revisions of "Part:BBa K274100"

Revision as of 06:26, 18 September 2009

CrtEBI with rbs

This Composite Biobrick is created by standard assembly of Part BBa_K118014 (CrtE with rbs), Part BBa_K118006 (CrtB with rbs) and Part BBa_K118005 (CrtI with rbs), which are submitted by previous iGEM teams. The whole cassette is on plasmid pSB1A2 (high copy, Ampicillin resistance).

Together, enzymes CrtE, CrtB and CrtI convert colourless farnesyl pyrophosphate to red lycopene (via intermediates geranylgeranyl pyroiphosphate and phytoene). The red lycopene pigment can then be used as a coloured signal output, e.g. for biosensors.

There is already individual ribosome binding site before each enzyme gene sequence. Internal restriction sites have been removed by previous iGEM teams. Please refer to Parts BBa_K118014, BBa_K118006 and BBa_K118005 for more information on individual Biobricks components.

This Composite Biobrick has been put under constitutive promoter R0011 (see Part BBa_K274110) and arabinose-inducible promoter I0500 (see Part BBa_274120), transformed and tested in E.coli strain MG1655.

Amount of lycopene produced can be measured by photospectrometer with absorbance at 475nm (lycopene extraction using acetone).

A related Composite Biobrick is Part BBa_K274200 (CrtEBIY with rbs), which "goes on" one step after lycopene, converting red lycopene to orange beta-carotene pigment.

Sequence and Features