Coding

Part:BBa_K3054015

Designed by: Dharmik Hitesh Patel   Group: iGEM19_Chalmers-Gothenburg   (2019-10-20)


bphA1 protein from Rhodococcus jostii with 6 His tag

Rhodococcus jostii was found to degrade lightly chlorinated PCB via a mutated version of biphenyl pathway. Codon optimized to express in yeast.


Usage and Biology

IGEM pathway.jpeg

Us, iGEM Chalmers Gothenburg, planned to engineer the following pathway to degrade a large spectrum of PCBs. A detailed pathway is shown in the diagram. This biobrick is a component of this pathway optimized to be expressed in yeast.


BphA protein

BphA is a multicomponent biphenyl dioxygenase system is composed of a ferredoxin reductase (BphA4), a ferredoxin (BphA3) and a terminal oxygenase (BphA1A2)

BphA1: Biphenyl 2,3-dioxygenase subunit alpha


Source:UniProtKB - Q53122 (BPHA1_RHOJR)


Expression of the biobrick

Western Blot

Results of western blot of His-tagged proteins. From left to right the samples, together with their His-tagged genes are WT, WT, 1 (bphA1), ladder, 2 (bphA4), 3 (bphC), 4 (bphD), positive control

The biobrick has been verified by sequencing and the protein was expressed in yeast using our dual promoter system.










Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 738
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 580
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 409


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Categories
Parameters
None