Part:BBa_K2587009

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