Difference between revisions of "Part:BBa K934001:Experience"

Revision as of 09:26, 25 September 2012

phaC1-A-B1 [P(3HB) synthesis]

To synthesize PHB by E.coli, we transformed E.coli JM109 with the constructed phaC1-A-B1 parts on pSB1C3 (BBa_K934001). E.coli JM109 is used to synthesize PHB, because it tends to have a high density accumulation of PHB(ref.5). As a negative control, we transformed E.coli JM109 with PlasI-gfp on pSB1C3.

FIG1 is the photographs of E.coli colonies on Nile red positive medium taken under UV. The orange colonies in FIG1.A show that the accumulated poly-3-hydroxybutyrate, PHB in cells was stained by Nile red. This result indicates that part BBa_K934001 synthesized PHB. FIG1.B is the photograph of negative control cells. In this figure we observed that there were no remarkable colored colonies.

FIG1.A: E.coli JM109 colonies with BBa_K934001 gene, PHB accumulation. FIG1.B: E.coli JM109 colonies with PlasI-gfp gene, no PHB accumulation.

FIG2 shows the difference between cells storing PHB and those not storing PHB. The cells in blue rectangle area are the cells with PHB synthesis gene and the cells in green rectangle area are the cells with plasI-gfp gene as a negative control.

FIG2 Difference between cells storing PHB and cells not storing PHB. Blue rectangle: with BBa_K934001 gene, PHB accumulationGreen rectangle: with PlasI-gfp gene, no PHB accumulation.

Using the cells storing PHB, we drew a rose silhouette on the LB agar plate containing Nile red. (FIG3).

FIG.3 Rose silhouette on the LB agar plate containing Nile red.

For more information, see [http://2012.igem.org/Team:Tokyo_Tech/Projects/PHAs/index.htm#3. our work in Tokyo_Tech 2012 wiki].

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