Part:BBa_K1686045

crdA gene with codon optimisation for E. coli

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 399
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 859
  Illegal AgeI site found at 828
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

Curdlan is a bacterial polysaccharide (linear (1->3)-β-glucan) that has been of significant recent interest due to its valuable properties and its bioactivity. In fact, Curdlan belongs to the class of biological response modifiers that enhance or restore normal immune defenses. For example, it can have antitumor, anti-infective, anti-inflammatory, and anticoagulant activities (see other properties of Curdlan). In particular, this (1->3)-β-glucan can stimulate plants' immune system.
Our project is focused on the production of curdlan for stimulate plant defense system against Downy Mildew.

The molecular genetics of curdlan production have been investigated in Agrobacterium sp. ATCC31749. Studies identified three genes (crdA, crdS and crdC) that are essential for curdlan production The crdASC genes occupy a contiguous 4,948 bp region of the genome ( the crdS gene is flanked by the crdA and crdC genes).

CrdA might assist translocation of the nascent polymer of Curdlan across the cytoplasmic membrane.

Note : This protein has not really been well described in literature.

For our project, we wanted to produce curdlan by E.coli, so this part has been made with a codon optimization of the natural crdA sequence for E.coli.

References

Stasinopoulos, S. J., Fisher, P. R., Stone, B. A., & Stanisich, V. A. (1999). Detection of two loci involved in (1-->3)-beta-glucan (curdlan) biosynthesis by Agrobacterium sp. ATCC31749, and comparative sequence analysis of the putative curdlan synthase gene. Glycobiology, 9(1), 31–41. https://doi.org/10.1093/glycob/9.1.31