Difference between revisions of "Part:BBa K633002"

Revision as of 00:52, 19 October 2011

RBS+signal peptide phoA+Cellulase

This part consists of a ribosome binding site (BBa_0034), along with a gene segment encoding the leader sequence of alkaline phosphatase, phoA, and the cellulase CelD (BBa_K63000). Since the objective of our project was to display chimeric proteins in the cell surface of Escherichia coli, a signal peptide was needed to translocate the protein into the periplasm. Alkaline phosphatase PhoA normally cannot fold properly in the cytoplasm due to a necessity of disulfide formation (Richeter et al., 2005), so the signal sequence phoA can be used to send bacterial periplasmic proteins across the membrane. Becker et al. (2005) reported that an efficient translocation of the protein membrane EstA through the E. Coli cytoplasmic membrane was achieved by replacing the native EstA sequence with phoA. It is expected then that CelD will be excreted into the extracellular medium using this part alone, or be displayed on the microbial cell surface, when fused with EstA membrane protein (BBa_K63001).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1908
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 721
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 585
Illegal AgeI site found at 228
Illegal AgeI site found at 644
Illegal AgeI site found at 1473
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K633002 short</partinfo>
-We used Rybosome Binding Site BBa_B0034 designed by by Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr, evaluated by Warsaw's 2010 team. we added membrane protein signal peptide phoA and cellulase immediately next to it
+This part consists of a ribosome binding site (BBa_0034), along with a gene segment encoding the leader sequence of alkaline phosphatase, phoA, and the cellulase CelD (BBa_K63000).
+Since the objective of our project was to display chimeric proteins in the cell surface of Escherichia coli, a signal peptide was needed to translocate the protein into the periplasm. Alkaline phosphatase PhoA normally cannot fold properly in the cytoplasm due to a necessity of disulfide formation (Richeter et al., 2005), so the signal sequence phoA can be used to send bacterial periplasmic proteins across the membrane. Becker et al. (2005) reported that an efficient translocation of the protein membrane EstA through the E. Coli cytoplasmic membrane was achieved by replacing the native EstA sequence with phoA. It is expected then that CelD will be excreted into the extracellular medium using this part alone, or be displayed on the microbial cell surface, when fused with EstA membrane protein (BBa_K63001).
 <!-- Add more about the biology of this part here