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Nucleic Acids Research, 1982, Vol. 10, No. 7 2399-2414
© 1982

MOLECULAR BIOLOGY

Laser crosslinking of E.coli RNA polymerase and T7 DNA

Catherine A. Harrison*, Douglas H. Turner* and David C. Hinkle+

*Department of Chemistry, University of Rochester Rochester, NY 14627, USA +Department of Biology, University of Rochester Rochester, NY 14627, USA

Received February 1, 1982. Accepted March 3, 1982.

The first photochemical crosslinking of a protein to a nucleic acid using laser excitation is reported. A single, 120 mJ, 20 ns pulse at 248 nm crosslinks about 10% of bound E. coli RNA polymerase to T7 DNA under the conditions studied. The crosslinking yield depends on mercaptoethanol concentration, and is a linear function of laser intensity. The protein subunits crosslinked to DNA are ß, ß' and {delta}.


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