Coding

Part:BBa_K078007:Design

Designed by: Sally.Wang (Jingyi Wang)   Group: iGEM08_Beijing_Normal   (2008-10-20)

2,3-dihydroxy-2,3-dihydrophenylpropionate dehydrogenase, the second step enzyme in PCBs degradation


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 443
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We use this part to construct the degradation pathway of PCBs.

2,3-dihydroxy-2,3-dihydrophenylpropionate dehydrogenase is coded by bphB.This is the second step enzyme of the PCBs deagradation pathway.

This is a standard parts. Acoording to the standard, we add EcoR1,Not1 and Xba1 upstream; and spe1, Not1, Pst1 and stop condon downstream. Anyone of the five sites could be used. The vector is the standard vector, pSB1AC3, provided by biobrick08. We recommend double digestion of EcoRI+pstI.Digest in NEB Buffer EcoRI + BSA at 37°C for about 3h.

Source

From the PCBs-degrading bacterium Burkholderia xenovorans LB400 chromosome 3. Type: genomic DNA


References

The biphenyl/polychlorinated biphenyl-degradation locus (bph) of Pseudomonas sp. LB400 encodes four additional metabolic enzymes. Bernd Hofer, Silke Backhaus and Kenneth N. Timmis.Gene, 144 (1994) 9-16