8/6/2019 ant Human RAD51 - OM-0215

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Potential uses of recombinant human RAD51

Biochemical and biological assays:• For DNA strand exchange• Protein-protein/protein-DNA interaction studies in vitro and in vivo • Western blots

Dr Luca Pellegrini and colleagues at Cambridge University have developed a methodfor preparing large quantities of purified recombinant full length human RAD51 whichis available for licensing.

RECOMBINANT HUMAN RAD51

For further information please contact:

Dr Rachel Atfield rachel.atfield@enterprise.cam.ac.uk

+44 (0)1223 760339

Cambridge Enterprise Limited, University of CambridgeHauser Forum, 3 Charles Babbage Road, Cambridge CB3 0GTUK

www.enterprise.cam.ac.uk

Case Ref: OM-0215

8/6/2019 ant Human RAD51 - OM-0215

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Figure 1: Recombinant human RAD51 Background

RAD51 is a 339 amino acid protein that is acritical component of the double-stranded breakpathway of DNA repair by homologousrecombination. Its important biological role derivesfrom its enzymatic ability to form nucleoproteinfilaments on single-stranded DNA that perform thestrand-exchange reactions of homologousrecombination. RAD51 is highly conserved inmost eukaryotes and is an essential protein invertebrates. Knockout mice are embryonic lethaland cells lacking functional RAD51 suffer

chromosome loss.

Technology

Dr Luca Pellegrini and colleagues in theDepartment of Biochemistry at the University ofCambridge have developed a method for rapidlypreparing recombinant full length human RAD51from bacteria. This enables the team to producelarge amounts of soluble, highly pure humanRAD51 (Figure 1). The team can typically producebetween 5 and 10 milligrams of pure RAD51 inone litre of bacterial culture using no protein tagssuch as His-tag, GST or MBP for the purificationprocess.

The recombinant human RAD51 produced by thismethod can be used in biochemical and biologicalassays for example for research into DNA repairby homologous recombination by performing DNAstrand-exchange assays for which RAD51 is anessential reagent. The RAD51 protein may alsoused in protein-protein and protein-DNAinteraction studies both in vitro (Figure 2) and in

vivo , as well as for use as a control in westernblots.

Commercialisation

We are seeking commercial partners for licensingsoluble recombinant human RAD51 protein.

OM-0215

DNA binding of RAD51 as shown by the change influorescence anisotropy of an Alexa488-labelled DNAmolecule in the presence of increasing amounts ofRAD51

SDS-PAGE analysis of purified, recombinanthuman RAD51

Figure 2: DNA-binding activity of recombinanthuman RAD51