Part:BBa_K533003:Design
OmpA-HIV aspartyl protease
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 936
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We need a protease to release our binding cassette in our E. coli transportation system. HIV protease has a high specificity and a relatively simple structure, which suits our application well.
We want to build an E. coli strain, expressing protease on its surface so that when it comes across another protein with the protease site, the protease can make the cut. Therefore, we fused the protein to the C-terminal of part of OmpA protein.
The whole coding sequence is under T7 promoter and lacI repressor. When expressed in BL21 (DE3) strain, the best condition for expression is 0.5mM IPTG induced at 18 centigrade for 12 hours.
Source
HIV protease sequence is from former part BBa_I712667. OmpA sequence is from BL21(DE3) e. Coli strain.