Difference between revisions of "Part:BBa K165007:Experience"
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== SDS-page and Western blot == | == SDS-page and Western blot == | ||
| − | [[Image:Gli1CFPproduction.PNG]] | + | [[Image:Gli1CFPproduction.PNG|left|x327px]] |
===Applications of BBa_K165007=== | ===Applications of BBa_K165007=== | ||
Revision as of 16:17, 26 October 2010
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Gli1 belongs to zinc finger family of proteins. Zinc fingers are small protein structural motifs that can coordinate one or more zinc ions to help stabilize their folds. They can be classified into several different structural families and typically function as DNA binding proteins. Zinc fingers coordinate zinc ions with a combination of cysteine and histidine residues and can be classified by the type and order of these zinc coordinating residues. Gli1 is composed of five tandem fingers that bind to DNA sequence: GAC CAC CCA AGA CGA. For isolation, purification and other experiments we added HIS tag to N-terminal side of Gli1 sequence and N-terminal split CFP on C-terminal part of Gli1 after an extension linker. We deposited modified variant of Gli1 as Gli1_link_nCFP (BBa_K323016) and additionally characterized it with several methods, including SDS-page, Western Blot and circular dichroism spectroscopy. In vitro binding of Gli1 to target DNA sequence was determined with surface plasmon resonance (SPR) and in vivo with betagalactosidase assay as a reporter.
SDS-page and Western blot
Applications of BBa_K165007
User Reviews
UNIQc3d5e8a6c42ae1f0-partinfo-00000000-QINU UNIQc3d5e8a6c42ae1f0-partinfo-00000001-QINU