Difference between revisions of "Part:BBa K243010"

Revision as of 20:06, 21 October 2009

His-FluA-Split Linker-Fok_i

This part unites some of our basic parts. The combination of a Histidin-Tag, an Anticalin-Tag and the linked protein domain Fok_a makes it possible to purify and detect the function of the protein. The fluoresceinA tag allows the measurement by quenching and the coupling to a fluorescein linked oligo.

Usage and Biology

This composite part is one part of our universal endonuclease and it needs another composite part like BBa_K243017 to built to a functional heterodimer. The FluA tag leads the part to DNA which is hybridized with a fluorescein labeled oligonucleotide. The Split Linker creates a distance of 51bp between the FluA and the linked Fok_i protein domain. The HisTag serves as a purification tag for Histidin column purification.

Purification

We purified this part of our universal endonuclease by Histidin column purification and proved the expression of the protein with a following Western Blot.

Western Blot: His-FluA-SplitLi-Fok_i in pEx;
lanes: NEB prestained marker broad range,
pool of elution fractions 2-5, empty lane, 3 positive controls

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 272
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K243010 short</partinfo>
-This part unites some of our basic parts. The combination of a [https://parts.igem.org/wiki/index.php?title=Part:BBa_K157011 Histidin-Tag], a [https://parts.igem.org/wiki/index.php?title=Part:BBa_K157004 Anticalin-Tag] and the [https://parts.igem.org/wiki/index.php?title=Part:BBa_K157009 linked] protein domain Fok_a, makes it possible to purifier and detect the function of the protein. The [https://parts.igem.org/wiki/index.php?title=Part:BBa_K157004 fluoresceinA tag] allows the measurement by quenching and the coupling to an flurescein linked oligo.<br>
+This part unites some of our basic parts. The combination of a [https://parts.igem.org/wiki/index.php?title=Part:BBa_K157011 Histidin-Tag], an [https://parts.igem.org/wiki/index.php?title=Part:BBa_K157004 Anticalin-Tag] and the [https://parts.igem.org/wiki/index.php?title=Part:BBa_K157009 linked] protein domain Fok_a makes it possible to purify and detect the function of the protein. The [https://parts.igem.org/wiki/index.php?title=Part:BBa_K157004 fluoresceinA tag] allows the measurement by quenching and the coupling to a fluorescein linked oligo.<br>
 ===Usage and Biology===
-This composite part is one part of our universal endonuclease and it needs another composite part like BBa_K243017 to built to a functional heterodimer. The FluA tag leads the part to DNA which is hybridized with an flurescein labeled oligonucleotide. The Split Linker mades a distance from 51bp between the FluA and the linked Fok_i protein domain. The HisTag serve as purification tag for Histidin column purification.
+This composite part is one part of our universal endonuclease and it needs another composite part like BBa_K243017 to built to a functional heterodimer. The FluA tag leads the part to DNA which is hybridized with a fluorescein labeled oligonucleotide. The Split Linker creates a distance of 51bp between the FluA and the linked Fok_i protein domain. The HisTag serves as a purification tag for Histidin column purification.
 ===Purification===
-We purified this part of our universal endonuclease by a Histidin column purification and proved the expression of the protein with a following Western Blot.<br>
+We purified this part of our universal endonuclease by Histidin column purification and proved the expression of the protein with a following Western Blot.<br>
 <br>
 [[Image:WB09.10.02jpg.jpg]]<br>