Coding

Part:BBa_K2587009

Designed by: Susanne Vollmer   Group: iGEM18_Duesseldorf   (2018-09-18)
Revision as of 09:19, 13 October 2018 by Suvol101 (Talk | contribs)


ptxD_opt

Most of the microorganisms, especially common used model organsims like Escherichia coli or Saccharomyces cerevisiae grow only very slow on phosphite as phosphorus source. Moreover in industry is not widely accepted to use antibiotic resistances as markers. Therefore a system is required, which allows avoidance of contamination by other microorganisms and at the same time represents a reliable selection marker. We present the use of the ptxD gene from Pseudomonas stutzeri together with a phosphite media (reduces growth of contaminants), which could abolish the use of antibiotics in the future and the more efficient use of a phosphorus source in the medium. In this case ptxD is codon optimised for S.cerevisiae.


Usage and Biology

  • ptxD codes for phosphonate dehydrogenase
  • oxidation of phosphite (phosphonate) using NAD+ and H20 to phosphate and NADH
  • Selection marker for budding and fission yeast
  • phosphite-oxidizing ability
  • environmentally safe culture
  • antibiotic free system
  • pH optimum: 7.25- 7.75

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 93
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 6
    Illegal BamHI site found at 1044
    Illegal XhoI site found at 216
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 714
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 22
    Illegal BsaI.rc site found at 1051

References

Kanda, Keisuke, et al. "Application of a phosphite dehydrogenase gene as a novel dominant selection marker for yeasts." Journal of biotechnology 182 (2014): 68-73.:
https://www.sciencedirect.com/science/article/pii/S016816561400193X

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Categories
//awards/basic_part/nominee
Parameters
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