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URL: http://www.addgene.org/103000
Proper Citation: RRID:Addgene_103000
Insert Name: PfSSB
Organism: Other
Bacterial Resistance: Ampicillin
Defining Citation: PMID:29466468
Vector Backbone Description: Vector Backbone:pET22B; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: The expressed protein is a Plasmodium falciparum single-stranded DNA-binding protein, PfSSB, for subsequent labeling at the single, native cysteine with one IDCC (N-[2-(iodoacetamido)ethyl]-7-diethylaminocoumarin-3-carboxamide) per subunit. This adduct is then used as a fluorescent single-stranded DNA (ssDNA) biosensor, having ~20-fold fluorescence increase on binding ssDNA with very high affinity. PfSSB was expressed without the N-terminal apicoplast localisation sequence (76 amino acids) but amino acids are numbered for the full-length protein, that is the first amino acid in the expressed protein is M77. Protocol See Hedgethorne and Webb (2012), which is also followed for PfSSB purification, except with the omission of the PEI precipitation step. Hedgethorne, K.; Webb, M. R., Fluorescent SSB as a reagentless biosensor for single-stranded DNA. Methods Mol. Biol. 2012, 922, 219-233.
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Source: Addgene
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