Part:BBa_K3394006
AIDA-I auto-display system
Since cellulose cannot cross the membrane, we engineered E. coli to express the cellulolytic complex on its surface through AIDA-1 auto-display system.
The AIDA-1 auto-display system we chose, utilizes the translocation mechanism of passenger proteins across the outer membrane through a C-terminal beta-barrel and its preceding linking region. We planned to use JK321 E. coli strain which is a DsbA-negative mutant strain of E. coli that facilitates the autodisplay of passenger proteins and contains the AIDA-1 auto display plasmid. The ompT-gene (Outer membrane protease T) in the DsbA-negative strain was mutated in order to obtain a stable surface display of the recombinant passenger proteins.
In figure 1, we can see the structure of the polyprotein precursor. By the use of a typical signal peptide (SP), a precursor protein is transported across the inner membrane. After arrival at the periplasm, the C-terminal part of the precursor folds as a porin-like structure, a so-called beta-barrel within the outer membrane, and the passenger proteins are transmitted to the cell surface.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 241
Illegal EcoRI site found at 937
Illegal EcoRI site found at 4139
Illegal PstI site found at 289
Illegal PstI site found at 875
Illegal PstI site found at 3387 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 241
Illegal EcoRI site found at 937
Illegal EcoRI site found at 4139
Illegal PstI site found at 289
Illegal PstI site found at 875
Illegal PstI site found at 3387 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 241
Illegal EcoRI site found at 937
Illegal EcoRI site found at 4139
Illegal BamHI site found at 1139 - 23INCOMPATIBLE WITH RFC[23]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 241
Illegal EcoRI site found at 937
Illegal EcoRI site found at 4139
Illegal PstI site found at 289
Illegal PstI site found at 875
Illegal PstI site found at 3387 - 25INCOMPATIBLE WITH RFC[25]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 241
Illegal EcoRI site found at 937
Illegal EcoRI site found at 4139
Illegal PstI site found at 289
Illegal PstI site found at 875
Illegal PstI site found at 3387
Illegal NgoMIV site found at 1351
Illegal NgoMIV site found at 1398
Illegal NgoMIV site found at 1766
Illegal NgoMIV site found at 1926
Illegal NgoMIV site found at 2280 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI.rc site found at 3213
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