Difference between revisions of "Part:BBa K1991009"
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<partinfo>BBa_K1991009 short</partinfo> | <partinfo>BBa_K1991009 short</partinfo> | ||
| − | + | === DISPLAYING AOX ENZYME ON THE CELL SURFACE OF E. COLI === | |
<p>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 ([https://www.ncbi.nlm.nih.gov/pubmed/11118595 Enzyme Microb Technol. 2001]). Bacillus lipase ([https://www.ncbi.nlm.nih.gov/pubmed/25163839 J Microbiol. 2014]) and Fungi xylanase ([https://www.ncbi.nlm.nih.gov/pubmed/25691337 Curr Microbiol. 2015]) were demonstrated to be displayed on the cell surface of E. coli and maintained the functional enzyme activities. In iGEM 2016 MINGDAO's project, AOX (alcohol oxidase) was displayed on bacterial surface by fusing with Lpp-OmpA, which was proved by protein analysis, enzyme activity assay and TMB assay</p> | <p>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 ([https://www.ncbi.nlm.nih.gov/pubmed/11118595 Enzyme Microb Technol. 2001]). Bacillus lipase ([https://www.ncbi.nlm.nih.gov/pubmed/25163839 J Microbiol. 2014]) and Fungi xylanase ([https://www.ncbi.nlm.nih.gov/pubmed/25691337 Curr Microbiol. 2015]) were demonstrated to be displayed on the cell surface of E. coli and maintained the functional enzyme activities. In iGEM 2016 MINGDAO's project, AOX (alcohol oxidase) was displayed on bacterial surface by fusing with Lpp-OmpA, which was proved by protein analysis, enzyme activity assay and TMB assay</p> | ||
Revision as of 19:18, 19 October 2016
Pcons-RBS-LO-AOX2-His
DISPLAYING AOX ENZYME ON THE CELL SURFACE OF E. COLI
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. In iGEM 2016 MINGDAO's project, AOX (alcohol oxidase) was displayed on bacterial surface by fusing with Lpp-OmpA, which was proved by protein analysis, enzyme activity assay and TMB assay
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]