Difference between revisions of "Part:BBa K208002:Design"

Latest revision as of 16:23, 22 October 2009

Gene III Signal Peptide - Silver Fusion Compatible

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Design Notes

Silver-fusion compatible Gene III secretion tag in pSB3K3. There are no additional design parameters.

Source

To construct the GeneIII sequence, complimentary forward and reverse oligonucleotides were synthesized by Eurofins Operon. 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. This part was successfully constructed and sequenced.

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

@@ Line 10: / Line 10: @@
 ===Source===
-To construct the GeneIII sequence, complimentary forward and reverse oligonucleotides were synthesized by Eurofins Operon. 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 each sequence. These parts were then cut with EcoRI and SpeI and ligated into a BioBrick vector. Each of these parts were successfully constructed and sequenced.
+To construct the GeneIII sequence, complimentary forward and reverse oligonucleotides were synthesized by Eurofins Operon. 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. This part was successfully constructed and sequenced.
 ===References===