Part:BBa_K934001:Experience
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.
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.
Using the cells storing PHB, we drew a rose silhouette on the LB agar plate containing Nile red. (FIG3).
For more information, see [http://2012.igem.org/Team:Tokyo_Tech/Projects/PHAs/index.htm#3. our work in Tokyo_Tech 2012 wiki].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 916
Illegal BglII site found at 1741 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 222
Illegal NgoMIV site found at 293
Illegal NgoMIV site found at 893
Illegal NgoMIV site found at 1205
Illegal NgoMIV site found at 1484
Illegal NgoMIV site found at 2136
Illegal NgoMIV site found at 2158 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 4002
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