Difference between revisions of "Part:BBa K323042"

Revision as of 09:14, 27 October 2010

cYFP link ZNF HIVC under T7 promoter

[http://2010.igem.org/Team:Slovenia Team Slovenia 2010] further characterized Zif268-HIV DNA-binding domain from Registry. Check results on parts original page under Experience. Zif268-HIV DNA- binding protein 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. We renamed this part into zinc finger HivC (a mutation variant of Zif268 zinc finger) according to work done by Reynolds and co. (2003). HivC is composed of three fingers that bind to DNA sequence: GAT GCT GCA. cCFP_link_HivC was deposited as BBa_K323071 and cloned behind T7 promoter to ensure production in E.coli BL21(DE3)pLysS strain. Its binding characteristics were analysed with several methods, including mobility shift analysis, surface plasmon resonance (SPR) and in vivo test of binding using betagalactosidase as a reporter. All results were deposited back to original page of HivC.

Sequence and Features