Difference between revisions of "Part:BBa K2450401"

Revision as of 11:22, 21 October 2017

OmpA SpyTag sfGFP label

OmpA with SpyTag Labelled with sfGFP

Sequence and Features

Assembly Compatibility:

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 <!-- Add more about the biology of this part here
 ===Usage and Biology===
+This part was designed to target proteins to the outer membrane and then outer membranes. As pathways targeting proteins to the outer membrane are not well understood, proteins must be bound to the outer membrane by other methods. The protein OmpA is abundant in the membrane, and efficiently targeted there by the Omp leader sequence.
+The protein being targeted to the outer membrane is expressed as a fusion protein with spycatcher to bond it to OmpA in the outer membrane. The spytag/spycatcher system has been used to target proteins to the outer membrane by creating an isopeptide bond between the spytag and spycatcher domains. As spytag is very short it does not disrupt membrane proteins to the extent that a large fusion protein would, allowing it to fold properly in the membrane. This method has been shown to work by Alves et al in 'Protecting enzymatic function through directed packaging into bacterial outer membrane vesicles' 2016.
+sfGFP is used to show the protein has been successfully incorporated to the outer membrane. sfGFP is required as it is being targeted to the periplasm where GFP cannot fold.
+As this is being targeted to the outer membrane,
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 <span class='h3bb'>Sequence and Features</span>