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URL: http://www.addgene.org/78202
Proper Citation: RRID:Addgene_78202
Insert Name: parM (K33A D63C T174A T175N D224C C287A)
Organism: E. coli
Bacterial Resistance: Ampicillin
Defining Citation: PMID:20158267
Vector Backbone Description: Vector Backbone:pHis17; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: The plasmid is used to express the plasmid segregation protein, ParM, from E. col. The protein is modified for use as the scaffold of an ADP biosensor, ATR-ParM. Two cysteine mutations are for label attachment (D63C and D224C); there is a mutation to inhibit filament formation (K33A), two mutations to weaken ATP binding (T174A and T175N), a mutation to change the exposed cysteine (C287A) and a C-terminal His6 tag to aid purification. When labelled with two iodoacetamidotetramethylrhodamine (IATR) fluorophores, this biosensor has a fluorescence intensity change of up to 20-fold and can be used to measure ADP in the range of hundreds of nanomolar to ~100 μM, in the presence of up to millimolar ATP. The biosensor can also be used for measurement of other nucleoside diphosphates e.g. GDP, UDP and others.
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Source: Addgene
