Composite

Part:BBa_K2533053:Design

Designed by: kangyuan yu   Group: iGEM18_HUST-China   (2018-09-21)


RBS-mleS-RBS-lldP-RBS-ldhA-TT


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 212
    Illegal BglII site found at 1942
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 406
    Illegal NgoMIV site found at 529
    Illegal NgoMIV site found at 571
    Illegal NgoMIV site found at 904
    Illegal NgoMIV site found at 1075
    Illegal NgoMIV site found at 1698
    Illegal NgoMIV site found at 2064
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 936
    Illegal BsaI site found at 2435
    Illegal BsaI.rc site found at 388
    Illegal BsaI.rc site found at 676
    Illegal BsaI.rc site found at 1243
    Illegal BsaI.rc site found at 1588
    Illegal BsaI.rc site found at 2331
    Illegal BsaI.rc site found at 4313


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 to produce lactate. Also, considering the necessity of helping Rhodopseudomonas palustris to transport lactate out of the membrane, we need to add a lldP gene. TT is a common terminator.

Source

Lactococcus lactis subsp. lactis Ⅱ1403 & 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