Difference between revisions of "Part:BBa K2940010"

Revision as of 15:06, 21 October 2019

Immobilised dye-degrading peroxidase: Dyp (without signal peptides)+ Cytochrome b5

This part is designed for the DyP-Cytb5 expression. So the signal peptides were removed from the original coding sequence (NC_000964.3) to make sure DyP can anchor with Cytb5. The strong constitute promoter and RBS combination (BBa_K608002) were inserted prior to each start codon. This part is related to the ghost shells project, which aims at immobilising DyP from Bacillus subtilis to the inner surface of the cytosolic membrane of Escherichia coli with the help of the C-terminal membrane anchor protein Cytb5. Then, the expression of Lysis protein E from PhiX174 can cause the formation of a pore in the cell wall of E. coli, which results in the release of the cytosol. Theoretically, the empty cellular envelopes with immoblised DyP can be used for degradation.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 995
Illegal SapI site found at 425
Illegal SapI site found at 783
Illegal SapI.rc site found at 939

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 <partinfo>BBa_K2940010 short</partinfo>
-This part is designed for the DyP-Cytb5 expression. So the signal peptides were removed from the original coding sequence to make sure DyP can anchor with Cytb5. The strong promoter and RBS were introduced to make the DNA construct. It was cloned into pSB1C3 using classcial cloning and was verified using ABTS assay
+This part is designed for the DyP-Cytb5 expression. So the signal peptides were removed from the original coding sequence (NC_000964.3) to make sure DyP can anchor with Cytb5. The strong constitute promoter and RBS combination (BBa_K608002) were inserted prior to each start codon. This part is related to the ghost shells project, which aims at immobilising DyP from Bacillus subtilis to the inner surface of the cytosolic membrane of Escherichia coli with the help of the C-terminal membrane anchor protein Cytb5. Then, the expression of Lysis protein E from PhiX174 can cause the formation of a pore in the cell wall of E. coli, which results in the release of the cytosol. Theoretically, the empty cellular envelopes with immoblised DyP can be used for degradation.
 <!-- Add more about the biology of this part here