Difference between revisions of "Part:BBa K243025"

Revision as of 14:27, 21 October 2009

Strep-FluA-Middle Linker-Fok_i

This combination of parts use the benefits of an Streptavidin-tag for purification. It is also linked with a FluroescineA-tag. The Middle Linker (GlySerGlyGly)x2 connects the parts and adds additional space between them to guarantee the independent function of FluA tag and the protein domain Fok_a.

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 FluroescineA-tag allows the measurement by quenching and the coupling to an flurescein linked oligo. To avoid interactions between the FluA-tag with the connected protein domain Fok_a we applied the Middel Linker. The Linker itself has no influence of the connected parts. We decided to use the Middle Linker for this construct to prove the optimal distance between the parts. It is important that the used linker has a certain flexibility and is long enough to avoid steric interferences between the parts. If the linker is too long it might cause a instability of the whole construct. Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 278
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K243025 short</partinfo>
-This combination of parts use the benefits of an Strep-tag and is linked with a FluroescineA tag. The MiddleLinker (GlySerGlyGly)x2 produce a certain distance between the FluA tag and the protein domain Fok_i.
-<!-- Add more about the biology of this part here
+This combination of parts use the benefits of an Streptavidin-tag for purification. It is also linked with a FluroescineA-tag. The Middle Linker (GlySerGlyGly)x2 connects the parts and adds additional space between them to guarantee the independent function of FluA tag and the protein domain Fok_a.
 ===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 FluroescineA-tag allows the measurement by quenching and the coupling to an flurescein linked oligo. To avoid interactions between the FluA-tag with the connected protein domain Fok_a we applied the Middel Linker. The Linker itself has no influence of the connected parts. We decided to use the Middle Linker for this construct to prove the optimal distance between the parts. It is important that the used linker has a certain flexibility and is long enough to avoid steric interferences between the parts. If the linker is too long it might cause a instability of the whole construct.
 <!-- -->
 <span class='h3bb'>Sequence and Features</span>