Difference between revisions of "Part:BBa K2382004:Design"
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
−
We have remove the stop coden of Thioredoxin
+
Thioredoxin could help the protein folding correctly, and make the fusion proteins in soluble form that are biologically active.And we remove the stop coden of Thioredoxin
−
(NC_000913.3) so that it can be used in fusion protein designing.
+
(NC_000913.3) so that it can be used in fusion protein designing.
−
 
+
We Insert linker between our two proteins: thioredoxine and MSMEG5998.
−
 
+
The purpose is to separate these two proteins
−
 
+
The sequence of Linker contain the polylinker ,and that is: GGTACCCGGGGATCCCTCGAGGGTGGT,
 +
There are four restriction site in it:
 +
Kpn1:GGT’ACC
 +
Sma1: CCC’GGG
 +
BamH1:GGA’TCC
 +
Xho1:C’TCGAG
 +
The last of the sequence plus two glycine, and it allow the protein behind linker can be reversed, reducing the probability of protein folding error.
  
 
===Source===
 
===Source===

Revision as of 12:13, 29 October 2017


Thioredoxin with polylinker


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 367
    Illegal XhoI site found at 373
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Thioredoxin could help the protein folding correctly, and make the fusion proteins in soluble form that are biologically active.And we remove the stop coden of Thioredoxin (NC_000913.3) so that it can be used in fusion protein designing. We Insert linker between our two proteins: thioredoxine and MSMEG5998. The purpose is to separate these two proteins The sequence of Linker contain the polylinker ,and that is: GGTACCCGGGGATCCCTCGAGGGTGGT, There are four restriction site in it: Kpn1:GGT’ACC Sma1: CCC’GGG BamH1:GGA’TCC Xho1:C’TCGAG The last of the sequence plus two glycine, and it allow the protein behind linker can be reversed, reducing the probability of protein folding error.

Source

Escherichia coli str. K-12 substr. MG1655 https://www.ncbi.nlm.nih.gov/nuccore/NC_000913.3

References