Team:Newcastle/glue

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ULB-Brussels developed BioBricks for production of Caulobacter crescentus glue, and won the Best New BioBrick Part, Natural award in 2009. Caulobacter crescentus is a Gram-negative bacterium, and the parts are optimised for E. coli. Unclear whether they will work in B. subtilis.

Genes: hfsE, hfsF, hfsG, hfsH, (hfsC), hfsI, hfsD, (hfsA, hfsB).

The inner membrane HfsE protein initiates glycosyltransferase by transferring N-acetylglucosamine (NAG) from UDP−NAG to a lipid carrier. HfsG, a second glycosyltransferase protein, transfers NAG subunits to the growing polysaccharide chain. HfsH deacetylates one or more NAG residues. The HfsF protein translocates the polysaccharide chain linked to the lipid carrier across the inner membrane. The polymerases HfsC and HfsI proteins link the NAG repeat units together. The holdfast polysaccharide is transferred across the outer membrane by HfsA, HfsB and HfsD proteins. Hfa proteins mediate the polysaccharide attachment to the cell.

Only hfsG and hfsH were inserted, since the other genes are already present in E. coli. Does B. subtilis have the other genes, or do we need to insert those too? Will it work at all in a Gram-positive bacterium?

References:

ULB-Brussels 2009 iGEM team. A new generation of glue.

Chris S. Smith, Aaron Hinz, Diane Bodenmiller, David E. Larson, and Yves V. Brun. 2002 Identification of Genes Required for Synthesis of the Adhesive Holdfast in Caulobacter crescentus