Composite

Part:BBa_K2533055:Design

Designed by: Ziyang Xiao   Group: iGEM18_HUST-China   (2018-09-28)


RBS-ldhA-RBS-lldP-TT


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1309
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 247
    Illegal NgoMIV site found at 457
    Illegal NgoMIV site found at 1065
    Illegal NgoMIV site found at 1431
    Illegal NgoMIV site found at 1503
    Illegal NgoMIV site found at 1572
    Illegal NgoMIV site found at 1706
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1802
    Illegal BsaI.rc site found at 982
    Illegal BsaI.rc site found at 1698


Design Notes

Compared with cyanobacteria, we find Rhodopseudomonas palustris is more suitable than cyanobacteria to provide Lactate for Shewenella due to its charactestics of Anaerobic respiration.Therefore, we decided to modify Rhodopseudomonas palustris so that it could produce lactate under anaerobic condition and transport lactate to the extracellular. And we have found that if we want to enhance the production of lactate from Rhodopseudomonas palustris, we can promote the convertion efficiency of pyruvate to D-lactate and malate to L-lactate. Therefore, we decided to use this two genes, mleS and ldhA, to help Rhodopseudomonas palustris produce lactate. Also, considering the necessity of helping Rhodopseudomonas palustris transport lactate out of the membrane, we need to add a lldP gene. In this part, we switch the composition of lldp and ldha to detect the production of Lactate. TT is a common terminator.

Source

Escherichia coli str. K-12 substr. MG1655 & Escherichia coli str. K-12 substr. MG1655

References

【1】：Henan journal of animal husbandry and veterinary medicine，by Wang yue xian ; Liu de hai 【2】：The Capacity of Photosynthesis Bacteria Rhodopseudomonas palustris Assimilating to Phosphate, Bulletin of Science and Technology, 2002, issue 2, pp 142-146 【3】：KEGG, https://www.genome.jp/kegg/pathway.html 【4】：Cloning and sequence analysis of the gene encoding Lactococcus lactis malolactic enzyme: relationships with malic enzymes FEMS Microbiol Lett. 1994 Feb 1;116(1):79-86 【5】：Fine tuning the transcription of ldhA for d-lactate production August 2012, Volume 39, Issue 8, pp 1209–1217 【6】：Transport of L-Lactate, D-Lactate, and Glycolate by the LldP and GlcA Membrane Carriers of Escherichia coli Volume 290, Issue 2, 18 January 2002, Pages 824-829 【7】：Enhancement of Hydrogen Production and Carbon Fixation in Purple Nonsulfur Bacterium Bacterium by Synthetic Biology Shou-Chen Lo 【8】：Jcat, http://www.jcat.de/#opennewwindow