Association Kinetics of DNA Oligomers

The general usefulness of RET in investigations of asso-ciadon reactions is illustrated by studies of donor and acc or-labeled DNA oligomas that have a conqilemai- 

RET has also been used to n asure the catalytic activity of restriction enzymes, which cleave DNA at specific sites in the sequence. One example of this applicatitm is on investigation of thePaeR endoom ease. For this lestiic tion enzyme the fecognftion file consists of a CTCOAG 

Flgure13.20. Prindiite of the RET endonucteMe assay. The emiteiiin apecOfumofa I6 iiKr loiioMiniid(—)ie quenched upon addition of its 

The concept used in the endonuclease assay was the dimination of RET by enzymatic breakage of a covalently linked D-A pair. This concept has been applied in avariety of other circumstances, including assay of an HIV-1 endonuclease, which is involved in integrating the HIV into the host DNA. RET has also been used to measure the activity of an HIV protease, proteolytic cleavage of linked derivatives of GEP, and the activity of ribozymes in cleaving RNA and as the basis of a general assay for DNA cleavage.  

Calculation of the transfer effidency from the enhanced acceptor emission requires careflil consideration of all the intenelaled intensities. Assuming that the donor does not emit at die acceptor wavelength, the eificieocy of transfer is given by 

---