Difference between revisions of "Part:BBa K243024"

Revision as of 14:07, 21 October 2009

Strep-FluA-Middle Linker-Fok_a

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 , 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 sterical interferences between the parts. If the linker is too long it might cause a instability of the whole construct.

Sequence and Features 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
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 1075

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 __NOTOC__
 <partinfo>BBa_K243024 short</partinfo>
 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
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 ===Usage and Biology===
-We used this combination , 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 decided to use the Middle Linker for this construct to prove the optimal distance between the parts.
+We applied the Streptavidine-tag to enable a , 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  sterical interferences between the parts. If the linker is too long it might cause a instability of the whole construct.
-This part is a linker, it can be used to connect two parts and add additional space between these parts. That can be necessary to avoid interactions between these parts. We used this part to connect our protein domains with our FluA and DigA domains, for our project the universal endonuclease.The sequence produced the aminoacids Gly-Gly-Ser-Gly-Gly-Gly-Ser-Gly .
-[edit] Usage and Biology
-Linker were used normally to connect two peptides, it is important for that connection,that the linker itself has no influence of the connected peptides. So the sequence of the linker is designed for amino acids which not interact with the environment. The amino acids glycine and serine are zwitterionic and hydrophile, these properties make them a good choice for the repetitive sequence of the linker.
-The length of the linker is important to guarantee the independent function of two connected parts. When the linker is too short there might be a sterical interference between the parts and when it is too long, it can cause an instability of the construct.Also it is important that the linker has a certain flexibility.
 Sequence and Features