Difference between revisions of "Part:BBa K243015"

Latest revision as of 02:15, 22 October 2009

Strep-DigA-Split Linker-Fok_i

This combination uses the benefits of a Streptavidin-tag for purification. It is also linked with a DigoxigeninA 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_i.

Usage and Biology

This composite part is one part of our universal endonuclease and it needs another composite part like BBa_K243024 to build a functional heterodimer. The DigA tag guides the part to DNA which is hybridized with a Digoxigenin labeled oligonucleotide. The Split Linker creates a distance of 51bp between the DigA and the linked Fok_i protein domain. The Strep Tag serves as a purification tag for Streptavidin column purification.

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 2: / Line 2: @@
 <partinfo>BBa_K243015 short</partinfo>
-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_i.
+This combination uses the benefits of a Streptavidin-tag for purification. It is also linked with a DigoxigeninA 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_i.
 ===Usage and Biology===
-This composite part is one part of our universal endonuclease and it needs another composite part like BBa_K243024 to built to a functional heterodimer. The DigA tag leads the part to DNA which is hybridized with an Digoxigenin labeled oligonucleotide. The split linker makes a distance from 51bp between the DigA and the linked Fok_i protein domain. The Strep-Tag serve as purification tag for Streptavidin column purification.
+This composite part is one part of our universal endonuclease and it needs another composite part like BBa_K243024 to build a functional heterodimer. The DigA tag guides the part to DNA which is hybridized with a Digoxigenin labeled oligonucleotide. The Split Linker creates a distance of 51bp between the DigA and the linked Fok_i protein domain. The Strep Tag serves as a purification tag for Streptavidin column purification.
-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 DigA-tag allows the coupling to an Digoxigenin linked oligo. Emanating from our 3D modeling this combination of DigA tagged oligo and the construct containing the protein domain Fok_i is not that efficiently than the use of a combination of FluA tagged oligo with a construct containing Fok_i. To avoid interactions between the DigA-tag with the connected protein domain Fok_i we applied the split Linker. This Linker is an improved part of the team Freiburg08 and it is a suitable linker for fusion proteins.
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