Difference between revisions of "Part:BBa K844001"

Revision as of 16:32, 2 October 2012

Spider Silk 1x 1E Subunit "U" (native sequence)

Spider silk subunit (1x) native sequence; contains one elasticity domain and one strength domain.

Usage and Biology

The sequence for this subunit was derived from the 1E silk subunit from Brooks et al. (2008), which is based on the native sequence of the MaSp2 silk gene in Argiope aurantia. The 1E silk subunit contains codons for only six amino acids in the following proportions: glycine (40%), alanine (17.5%), glutamine (15%), proline (15%), serine (7.5%), and tyrosine (5%). The native Argiope aurantia codon usage profile is significantly different from E. coli ’s and the GC% of the DNA molecule itself (75%) is significantly different from the average GC% of native E. coli genes (51%). These features can make the native form of the gene difficult and slow to produce for the ribosomes in the cell as the necessary tRNA molecules are present in lower concentrations in E. coli and the genetic machinery of E. coli is not adapted to opening DNA of high GC% when making mRNA, and thus lower amounts of spider silk mRNA are produced.

The silk subunit contains three main domains: an elasticity domain, a linker domain, and a strength domain. The elasticity domain contains sequences coding for beta-helices and beta-spiral in the protein; these structures increase the elasticity of the spider silk. The linker domain joins the strength and elasticity domains together. The strength domain will form beta-sheets and strengthen the silk fibers by cross-linking silk strands.

Related parts:

BBa_K844001 – the native spider silk subunit with two elasticity domains and one strength domain (no sequence manipulation)

BBa_K844005 – this spider silk subunit with an added Met (ATG) start codon at the 5’ end

BBa_K844009 – this spider silk subunit with an added 10-His Tag and double stop TAATAA sequence at the 3’ end

References:

Brooks AE, Stricker SM, Joshi SB, Kamerzell TJ, Middaugh CR, and Lewis RV. 2008. Properties of synthetic spider silk fibers based on Argiope aurantia MaSp2. Biomacromolecules 9:1506–10.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 19
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K844001 short</partinfo>
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 Spider silk subunit (1x) native sequence; contains one elasticity domain and one strength domain.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+The sequence for this subunit was derived from the 1E silk subunit from Brooks et al. (2008), which is based on the native sequence of the MaSp2 silk gene in Argiope aurantia.  The 1E silk subunit contains codons for only six amino acids in the following proportions: glycine (40%), alanine (17.5%), glutamine (15%), proline (15%), serine (7.5%), and tyrosine (5%). The native Argiope aurantia codon usage profile is significantly different from E. coli ’s and the GC% of the DNA molecule itself (75%) is significantly different from the average GC% of native E. coli  genes (51%). These features can make the native form of the gene difficult and slow to produce for the ribosomes in the cell as the necessary tRNA molecules are present in lower concentrations in E. coli and the genetic machinery of E. coli is not adapted to opening DNA of high GC% when making mRNA, and thus lower amounts of spider silk mRNA are produced.
+The silk subunit contains three main domains: an elasticity domain, a linker domain, and a strength domain. The elasticity domain contains sequences coding for beta-helices and beta-spiral in the protein; these structures increase the elasticity of the spider silk. The linker domain joins the strength and elasticity domains together. The strength domain will form beta-sheets and strengthen the silk fibers by cross-linking silk strands.
+Related parts:
+BBa_K844001 – the native spider silk subunit with '''two''' elasticity domains and one strength domain (no sequence manipulation)
+BBa_K844005 – this spider silk subunit with an added Met (ATG) start codon at the 5’ end
+BBa_K844009 – this spider silk subunit with an added 10-His Tag and double stop TAATAA sequence at the 3’ end
+References:
+Brooks AE, Stricker SM, Joshi SB, Kamerzell TJ, Middaugh CR, and Lewis RV. 2008. Properties of synthetic spider silk fibers based on Argiope aurantia MaSp2. Biomacromolecules  9:1506–10.
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