Part:BBa_K5175043:Design
T7 promoter-rhlA-rhlB-T7 terminator-T7 promoter-phaZ-T7 terminator
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 827
Illegal PstI site found at 1811
Illegal PstI site found at 1925 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2310
Illegal NheI site found at 2366
Illegal NheI site found at 3351
Illegal PstI site found at 827
Illegal PstI site found at 1811
Illegal PstI site found at 1925 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 176
Illegal BamHI site found at 736
Illegal XhoI site found at 912
Illegal XhoI site found at 2159 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 827
Illegal PstI site found at 1811
Illegal PstI site found at 1925 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 827
Illegal PstI site found at 1811
Illegal PstI site found at 1925
Illegal NgoMIV site found at 1052
Illegal NgoMIV site found at 1773
Illegal NgoMIV site found at 1886
Illegal NgoMIV site found at 2858
Illegal NgoMIV site found at 3059 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
It is a composite component consisting of the T7 promoter, T7 terminator, target genes rhlA, rhlB, phaZ. It is responsible for synthesizing β-hydroxy fatty acids and catalysing the synthesis of mono-rhamnolipids containing one rhamnosyl group from dTDP-L-rhamnose and β-hydroxy fatty acids, and inhibiting PHA anabolic bypass increases rhamnolipid production. RhlA encoded by the rhlA gene is a phthalyltransferase responsible for the synthesis of β-hydroxy fatty acids. RhlB encoded by the rhlB gene is responsible for catalyzing the synthesis of mono-rhamnolipids containing one rhamnose group from dTDP-L-rhamnose and β-hydroxy fatty acids. PhaZ encodes the poly(3-hydroxyalkanoate) depolymerase.
Source
rhlA, rhlB are from Pseudomonas aeruginosa.
phaZ is from Pseudomonas putida.
References
[1] PATRAUCHAN MARIANNA A, FLORIZONE C, DOSANJH M, et al. Catabolism of Benzoate and Phthalate in Rhodococcus sp. Strain RHA1: Redundancies and Convergence [J]. Journal of Bacteriology, 2005, 187(12): 4050-63.
[2] JARVIS F G, JOHNSON M J. A Glyco-lipide Produced by Pseudomonas Aeruginosa [J]. Journal of the American Chemical Society, 1949, 71(12): 4124-6.
[3] ZHU K, ROCK CHARLES O. RhlA Converts β-Hydroxyacyl-Acyl Carrier Protein Intermediates in Fatty Acid Synthesis to the β-Hydroxydecanoyl-β-Hydroxydecanoate Component of Rhamnolipids in Pseudomonas aeruginosa [J]. Journal of Bacteriology, 2008, 190(9): 3147-54.
[4] WITTGENS A, TISO T, ARNDT T T, et al. Growth independent rhamnolipid production from glucose using the non-pathogenic Pseudomonas putida KT2440 [J]. Microb Cell Fact, 2011, 10: 80.
[5] LIU H, CHEN Y, ZHANG Y, et al. Enhanced production of polyhydroxyalkanoates in Pseudomonas putida KT2440 by a combination of genome streamlining and promoter engineering [J]. International Journal of Biological Macromolecules, 2022, 209: 117-24.
[6] 尹进, 车雪梅, 陈国强. 聚羟基脂肪酸酯的研究进展 [J]. 生物工程学报, 32(6): 726-37.