Difference between revisions of "Part:BBa K1554002"

Revision as of 23:38, 15 October 2014

HarFAR

HarFAR-3 fatty acid reductase from Helicoverpa armigera with an ER retention signal (KKYR) attached to the C-terminal end. Codon optimization was perfomed for Nicotiana benthamiana.

Usage and Biology

The HarFAR protein is a fatty acid reductase coming from Helicoverpa armigera which transforms fatty acids into fatty alcohols.

Part:BBa_K1554001 (AtrΔ11), Part:BBa_K1554002 (HarFAR) and Part:BBa_K1554003 (EaDAcT) are enzymes of a biosynthesis pathway that lead to the production of insect sexual pheromones, Z11-16:OH and Z11-16:OAc, using palmitate as substrate.

Figure 1. Insect sexual pheromone pathway for Nicotiana benthamiana.

In our project we made a device with these three pheromones and expressed them by transient expression it in our plant chasis, Nicotiana benthamiana. In order to check if the insect sexual pheromones were present, we performed the analysis using HS-SPME coupled to GC-MS. We observed to additional peaks in the transformed plants that were not present in the control and have a similar mass spectrum and retention time as the standards, which confirmed that both molecules were the desired pheromones, (Z)-11-hexadecen-1-ol and (Z)-11-hexadecenyl acetate.

Figure 2. GC-MS analysis of the volatile organic compounds from a negative control of Nicotiana benthamiana.

Figure 3. GC-MS analysis of the volatile organic compounds from a genetically engineered Nicotiana benthamiana to produce insect pheromones.

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 1104
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 929

@@ Line 7: / Line 7: @@
 The HarFAR protein is a fatty acid reductase coming from ''Helicoverpa armigera'' which transforms fatty acids into fatty alcohols.
-Our team used it in with [https://parts.igem.org/Part:BBa_K1554001 Part:BBa_K1554001] to produce the insect pheromone Z11-16:OH in ''Nicotiana benthamiana'' using palmitate:CoA as precursor. It was also used with [https://parts.igem.org/Part:BBa_K1554003 Part:BBa_K1554003] to transform Z11-16:OH into another insect pheromone, Z11-16:OAc.
+[https://parts.igem.org/Part:BBa_K1554001 Part:BBa_K1554001] (Atr&Delta;11), [https://parts.igem.org/Part:BBa_K1554002 Part:BBa_K1554002] (HarFAR) and [https://parts.igem.org/Part:BBa_K1554003 Part:BBa_K1554003] (EaDAcT) are enzymes of a biosynthesis pathway that lead to the production of insect sexual pheromones, Z11-16:OH and Z11-16:OAc, using palmitate as substrate.
+[[File:VUPV_pathway.png]]
+Figure 1. Insect sexual pheromone pathway for ''Nicotiana benthamiana''.
+In our project we made a device with these three pheromones and expressed them by transient expression it in our plant chasis, ''Nicotiana benthamiana''. In order to check if the insect sexual pheromones were present, we performed the analysis using HS-SPME coupled to GC-MS. We observed to additional peaks in the transformed plants that were not present in the control and have a similar mass spectrum and retention time as the standards, which confirmed that both molecules were the desired pheromones, (Z)-11-hexadecen-1-ol and (Z)-11-hexadecenyl acetate.
+[[File:UPV_rutas-biosintesis_feromonas.png‎]]
+Figure 2. GC-MS analysis of the volatile organic compounds from a negative control of ''Nicotiana benthamiana''.
+[[File:VUPV_pheromone.png]]
+Figure 3. GC-MS analysis of the volatile organic compounds from a genetically engineered ''Nicotiana benthamiana'' to produce insect pheromones.
 <span class='h3bb'>Sequence and Features</span>