Part:BBa_K196004:Design

HfsG + HfsH proteins from Caulobacter crescentus

Caulobacter crescentus is an aquatic, Gram-negative bacterium that divides asymmetrically and is able to synthetize a strong glue. This glue is mainly made of a polysaccharide. There are different proteins needed to synthetize, export and attach it to the stalk of Caulobacter. To see the hole system, please see this page [http://jb.asm.org/cgi/content/full/190/21/7219/F8]. In our project, we would like this glue to be produced by Escherichia coli. As E. coli does have homolog genes for many of these proteins, but not for HfsG and HfsH, we decided to create a plasmid including only the genes coding for these two proteins. HfsG is a glycosyltransferase and HfsH is a carbohydrate esterase. Here you have a part made of the Biobricks HfsG (BBa_K196002) and HfsH (BBa_K196003).

Assembly Compatibility:

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

  Illegal BglII site found at 1470
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 480
  Illegal AgeI site found at 379
  Illegal AgeI site found at 864
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

As many mutations were needed to make the part compatible with the standard 10, we decided to make it synthetized by GeneArt [http://www.geneart.com]. We also optimized it for E. coli. We mean that the codons were changed to favour the most present in E. coli.

Source

This sequence comes from Caulobacter crescentus.

References

Toh, E, H. D. Kurtz, Jr., and Y.V. Brun. 2008. Characterization of the Caulobacter crescentus Holdfast Polysaccharide Biosynthesis Pathway Reveals Significant Redundancy in the Initiating Glycosyltransferase and Polymerase steps. Journal of Bacteriology, 190, 7219-7231.

Smith, C. S., A. Hinz, D. Bodenmiller, D. E. Larson, and Y. V. Brun. 2003. Identification of genes required for synthesis of the adhesive holdfast in Caulobacter crescentus. J. Bacteriol. 185:1342-1442.