Difference between revisions of "Part:BBa K243022"
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<partinfo>BBa_K243022 short</partinfo> | <partinfo>BBa_K243022 short</partinfo> | ||
| − | This combination uses the benefits of an Streptavidin-tag for purification. It is also linked with a | + | This combination uses the benefits of an Streptavidin-tag for purification. It is also linked with a DigoxigenineA-tag [https://parts.igem.org/wiki/index.php?title=Part:BBa_K243003 DigA]. The Split Linker connects the parts and adds additional space between them to guarantee the independent function of DigA-tag and the protein domain Fok_a.The DsbA tag is used to transfer the produced protein into the periplasm. |
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | + | We applied the Streptavidine-tag to enable a simultaneous purification of constructs with a His-tag. Strep-tag shows also a higher affinity towards Strep-Tactin than His-tag. For that the purification with Strep-tag is more specific. The used DigoxigeninA-tag allows the coupling to an flurescein linked oligo. Emanating from our 3D modeling this combination of DigA tagged oligos and the construct containing the protein domain Fok_a is more efficiently than the use of a combination of DigA tagged oligos with a construct containing Fok_i. To avoid interactions between the DigA-tag with the connected protein domain Fok_a we applied the Split Linker. The Linker itself has no influence of the connected parts. We decided to use the Split Linker for this construct to get a longer distance between DigA-tag and Fok_a to guarantee the independent function of both parts. this Linker is an improved part ove the optimal distance between the parts. It is important that the used linker has a certain flexibility and is long enough to avoid sterical interferences between the parts. If the linker is too long it might cause a | |
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Revision as of 19:44, 21 October 2009
DsbA-Strep-DigA-Split Linker-Fok_a
This combination uses the benefits of an Streptavidin-tag for purification. It is also linked with a DigoxigenineA-tag DigA. The Split Linker connects the parts and adds additional space between them to guarantee the independent function of DigA-tag and the protein domain Fok_a.The DsbA tag is used to transfer the produced protein into the periplasm.
Usage and Biology
We applied the Streptavidine-tag to enable a simultaneous purification of constructs with a His-tag. Strep-tag shows also a higher affinity towards Strep-Tactin than His-tag. For that the purification with Strep-tag is more specific. The used DigoxigeninA-tag allows the coupling to an flurescein linked oligo. Emanating from our 3D modeling this combination of DigA tagged oligos and the construct containing the protein domain Fok_a is more efficiently than the use of a combination of DigA tagged oligos with a construct containing Fok_i. To avoid interactions between the DigA-tag with the connected protein domain Fok_a we applied the Split Linker. The Linker itself has no influence of the connected parts. We decided to use the Split Linker for this construct to get a longer distance between DigA-tag and Fok_a to guarantee the independent function of both parts. this Linker is an improved part ove the optimal distance between the parts. It is important that the used linker has a certain flexibility and is long enough to avoid sterical interferences between the parts. If the linker is too long it might cause a Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 338
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1186