Part:BBa_K1037001

ygfD

botin-independent propionyl-CoA carboxylase;a succinate (or propionate) CoA ligase, or a novel (biotin-independent) propionyl-CoA carboxylase

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 743
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

1.we firstly send our plasmid to company for gene sequence. And do sequence alignment with data from NCBI to confirm that there is no other mutant aside of the mutation we do to remove restriction site and ORF is right. data link of NCBI:
http://www.ncbi.nlm.nih.gov/nuccore/NC_000913.2?report=genbank&from=3060859&to=3062152
2. We than induce the cell transformed with plasmid to see ygfD expression. Follow is the photo of SDS-PAGE gel analysis.

3.To prove the function of the biobrick further! We do fermentation and use HPLC to measure the increase of propionate.Follow is the results.

Conclusion : with all these result, we proved that our biobrick is well constructed . We can see both the protein expressed and prove the function of the protein.