Difference between revisions of "Part:BBa K2450401"
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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 [Alves et al 2016]. For an example of a part designed to be targeted to the outer membrane in this way see part [BBa_K2450501]. | 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 [Alves et al 2016]. For an example of a part designed to be targeted to the outer membrane in this way see part [BBa_K2450501]. | ||
| + | For more information on how this part was used in our project, please see our wiki: http://2017.igem.org/Team:Oxford | ||
| − | + | ===Sequence and Features=== | |
<partinfo>BBa_K2450401 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2450401 SequenceAndFeatures</partinfo> | ||
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| − | |||
===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K2450401 parameters</partinfo> | <partinfo>BBa_K2450401 parameters</partinfo> | ||
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Latest revision as of 23:00, 1 November 2017
OmpA SpyTag sfGFP label
OmpA with SpyTag Labelled with sfGFP
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. Its usual function is as an ion channel in the outer membrane.
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 [Alves et al 2016]. For an example of a part designed to be targeted to the outer membrane in this way see part [BBa_K2450501].
For more information on how this part was used in our project, please see our wiki: http://2017.igem.org/Team:Oxford
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 4
- 1000COMPATIBLE WITH RFC[1000]