Team:Cambridge/Project/ME02

From 2009.igem.org


Melanin Pigment


Design

Constructing the BioBrick

Biobrick

Our aim is to make the MelA gene into a biobrick as follows:

MelAbiobrick.jpg


Registry Code Type Sequence Description / Notes Length
BBa_K274001 Reporter 'MelA. The gene (melA) codes for a tyrosinase which produces a dark brown pigment from L-tyrosine. Production of the pigment requires the addition of copper and L-tyrosine supplements (the copper acts as a cofactor for the gene product) but no other precursors. The BioBrick sequence includes the native ribosome binding site. 1844bp


In order to do so, we had to remove forbidden restriction sites within the gene using primers and the in-fusion PCR technique. Primers were ordered as shown by the image below, to add the prefix and suffix and change the PstI restriction site from CTGCAG to CTGCAA:

MelA primer map.JPG

The PCR's will be done in the following steps, in order to remove both sites and prepare the gene as a biobrick:

MelA.png

Cambridge Sponsor Logo1.pngCambridge Sponsor Logo2.pngCambridge Sponsor Logo3.pngCambridge Sponsor Logo4.pngCambridge Sponsor Logo5.pngCambridge Sponsor Logo8.pngCambridge Sponsor Logo6.pngCambridge Sponsor Logo7.pngCambridge Sponsor Logo9.pngCambridge Sponsor Logo10.pngCambridge Sponsor Logo11.pngCambridge Sponsor Logo12.pngCambridge Sponsor Logo14.pngCambridge Sponsor Logo13.pngCambridge Sponsor Logo15.pngCambridge Sponsor Logo16.pngCambridge Sponsor Logo17.pngCambridge Sponsor Logo18.pngCambridge Sponsor Logo19.pngBmglab.jpg