*In our project Ompa-link is used as outer membrane anchor for our selection system
*In our project Ompa-link is used as outer membrane anchor for our selection system
*This brick contains our nonstandard restriction sites (NdeI and SacI) that allow 'scarless' cloning and easy creation of translation fusions
*This brick contains our nonstandard restriction sites (NdeI and SacI) that allow 'scarless' cloning and easy creation of translation fusions
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| [[Image:Ompalinker.jpg|200px]]
===Improvement by NEFU_China===
===Improvement by NEFU_China===
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Our team worked to improve the characterization of a signal peptide, BBa_K103006 , part from the iGEM Registry. This part originated from Univeristy of Warsaw 2008 iGEM team can be used as a outer-membrane-targeting anchor for selection system in E. coli.
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In order to make the production process and purification more efficiently, we added T7 promoter and His tag to the former of Lpp-ompa sequence and our another basic part (L-FABP) was fused to the back, constructing the composite parts (Lpp-ompa-L-FABP). Finally we linked the newly synthesized parts to the standard vector pSB1C3. The new plasmids can be used directly for IPTG induced expression in E. coli (DE3). See more details in composite part (BBa_K2302011)
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This biobrick (Lpp-ompA)(BBa_K2302003) is an improvement on the biobrick designed by the Warsaw 2008 iGEM team K103006 , and our improvement of this part is easy：
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1.The sequence that we have used is a synthetic gene with codon optimization, designed specifically for high level expression in E. coli. Thus it is shorter but with the same function as the other OmpA in the part registry.
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2. This part contains our nonstandard restriction sites (NdeI and Xhol) on the right and left separately of our newly Lpp-ompA that allows easy creation of translation fusion other interested protein.
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| [[Image:Ompalinker.jpg|200px]]
| [[Image:Ompalinker.jpg|200px]]
This protein structure was predicted using threading and protein fold prediction and may differ from actual one.
This protein structure was predicted using threading and protein fold prediction and may differ from actual one.
Revision as of 13:37, 26 October 2017
OmpA outer membrane protein A fused to linker; displays proteins on cell surface
One of our basic bricks used to create fusions attached to outer membrane
Usage and Biology
This part uses OmpA (outer membrane protein A [http://www.ncbi.nlm.nih.gov/pubmed/17559395?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum]) fragment (AAs 46-159) N-terminally fused with lipoprotein signal peptide fragment (AAs 1-9)
Proteins fused to Ompa-link are presented on cell's surface
Unstructuralized linker (Gly Ser Gly) allows proper folding of both fusion partners
In our project Ompa-link is used as outer membrane anchor for our selection system
This brick contains our nonstandard restriction sites (NdeI and SacI) that allow 'scarless' cloning and easy creation of translation fusions
Improvement by NEFU_China
This protein structure was predicted using threading and protein fold prediction and may differ from actual one.
