Team:Newcastle/glue

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Revision as of 01:14, 11 June 2010

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?

@@ Line 1: / Line 1: @@
 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''.
+''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
+Genes: hfsE, hfsF, hfsG, hfsH, hfsC, hfsI, hfsD, hfsA, hfsB.
-                                  The inner membrane HfsE protein initiates glycosyltransferase by transferring
+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.
-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.
-Complementary approaches are foreseen to differentiate these assumptions:
+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?
-  • The other genes (hfsE hsfF, hsfC hsfI, hsfD hsfA and hsfB) should also be transferred to
-    E. Coli with the same transcription rate.
-  • As the anchoring genes may play a role in the glue composition, hfaA, hfaB and hfaD
-    genes should be inserted in E.Coli too.
-  • We need to have more information about the adhesive composition. A proteomic ap-
-    proach with C.crescentus mutants that do not produce the glue might be undertaken.