Part:BBa_K1991009
Pcons-RBS-LO-AOX2-His
DISPLAYING AOX ENZYME ON THE CELL SURFACE OF E. COLI
Protein induction and purification may cost much time up to 12 hours and need tricky technical skills. To simplify the procedure of AOX enzyme application, we obtained the idea from the project of an iGEM Team – [http://2015.igem.org/Team:NCTU_Formosa/Design NCTU_Formosa] in 2015. A protein can be displayed on the cell surface of E. coli by fusing to Lpp-OmpA. Lipoprotein ([http://www.uniprot.org/uniprot/P69776 Lpp]) is a major outer membrane of E. coli which interacts with the peptidoglycan to maintain the structure and function of cell membrane. Another transmembrane protein called outer membrane protein A ([http://www.uniprot.org/uniprot/P0A910 OmpA]) is involved in bacterial conjugation and phage infection. A study showed that Lpp-OmpA hybrid can direct the heterologous protein GFP to the external surface of E. coli (Enzyme Microb Technol. 2001). Bacillus lipase (J Microbiol. 2014) and Fungi xylanase (Curr Microbiol. 2015) were demonstrated to be displayed on the cell surface of E. coli and maintained the functional enzyme activities. We’d like to display the AOX enzyme on bacterial surface by fusing with Lpp-OmpA and apply to the electrochemical analyzer by depositing on the test strip.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NotI site found at 2498 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 497
Illegal XhoI site found at 2507 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 452
- 1000COMPATIBLE WITH RFC[1000]
| None |