Part:BBa_K4680006

Roseovarius sp. 217 TMM Generator

Usage and Biology

The FSU iGEM team created this biological device to oxidize trimethylamine (TMA) in the small intestine of a human, helping those who are struggling with trimethylaminuria (TMAU). TMAU is a rare condition (or disease) in which a mutation appears on the FMO3 enzyme coding sequence and the body can no longer oxidize trimethylamine into trimethylamine N-oxide in the liver. This causes the person to expel a fishy odor throughout their entire body. Our device consists of an Anderson promoter, synthetic RBS, a TMM enzyme coding sequence, and a terminator. The TMM coding sequence comes from a species called Roseovarius sp 217. This species comes from the Marine Roseobacter clade that uses TMAO as a nutrient source. The Vmax and Km values of the TMM enzyme are 67.0 +/- 3.5 μg*min−1*mg−1 and 21.6 +/- 1.9 μM.

Measurements

This biological device has been inserted in a pIDTSmart-Kan plasmid backbone from IDT resulting in pFSUIGEM200. Figure 1 depicts the map of pFSUIGEM200. Roseovarius species 217 is a gram-negative bacteria of the phylum pseudomonadota. It was isolated from surface seawater collected near Plymouth, England. Due to its ability to oxidize TMA and its stronger enzymatic performance, the enzyme from it was utilized in our project. It is a biological device because it combines an anderson promoter for initiating transcription with the ribosome entry site that promotes initiation of protein synthesis with the enzyme itself and then a terminator to halt gene expression.

Figure 1 - pFSUIGEM200

Test

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 806
  Illegal PstI site found at 1397
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal PstI site found at 806
  Illegal PstI site found at 1397
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 806
  Illegal PstI site found at 1397
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 806
  Illegal PstI site found at 1397
  Illegal AgeI site found at 253
  
• 1000
  COMPATIBLE WITH RFC[1000]