Part:BBa_K208004:Design
PelB Signal Peptide - Silver Fusion Compatible
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 54
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
PelB Silver-fusion compatible secretion tag in pSB1A2. This part has been sequenced.
Another PelB BioBrick is available in the registry as BBa_J32015. However, we were unable to determine whether this part was Silver-fusion compatible. It was decided that a new version of this Biobrick be made to ensure Silver-fusion compatibility. Please refer to this website for more information: http://www.openwetware.org/wiki/Silver:_BB_Strategy
Source
To construct the OmpA sequence, complimentary forward and reverse oligonucleotides were synthesized. These strands were then annealed together. The oligonucleotides were designed so that the Silver fusion prefix and suffix sequences were appended onto the end of the sequence. This parts was then cut with EcoRI and SpeI and ligated into a BioBrick vector.
References
1. Choi JH, Lee SY (2004) Secretory and extracellular production of recombinant proteins using Escherichia coli Appl Microbiol Biotechnol 64:625-635
2. Luirink J, Oudega B (2004) Protein targeting to the inner membrane. In: Oudega B, editor. Kluwer Academic Publishers, pp 1-21
3. Molhoj M (2004) Leader sequences are not signal peptides. Nat Biotech 22:1502
4. Pugsley AP (1993) The complete general secretory pathway in gram-negative bacteria. Microbiol Rev 57:50-108
5. van der Does C, Noewen N, Driessen AJM (2004) The Sec translocase. In: Oudega B, editor. Kluwer Academic Publishers, pp:23-49
6. Veenendaal AKL, van der Does C, Driessen AJM (2004) The protein-conducting channel SecYEG. Biochimica et Biophysica Act 1694:81-95
7. von Heijne G (1986) Net N-C charge imbalance may be important for signal sequence function in bacteria. J Mol Biol 192:287-290