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DEVELOPMENT OF A PLAQUE REDUCTION ASSAY AND APPLICATION TO THE STUDY OF PSORALEN‐DAMAGED DNA
Author(s) -
Yokobata Kathy E.,
Jordan John M.,
Chapman Orvrlle,
Krerl Curt
Publication year - 1986
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1986.tb05620.x
Subject(s) - psoralen , virus quantification , chemistry , dna , bacteriophage , transfection , covalent bond , microbiology and biotechnology , biochemistry , biology , cell culture , organic chemistry , escherichia coli , genetics , gene
— We have developed a procedure called a plaque reduction assay to assess the biological activity of duplex circular DNA modified by covalent adduct formation with psoralen derivatives. The replicating form (RF) of bacteriophage DNA modified by photochemical addition of a psoralen derivative was introduced into bacterial cells using the CaCI 2 transfection method. The transfected cells. plated upon a confluent lawn of cells permissive for the bacteriophage in the inoculum, provided a measure of the reduction in infectivity of the RF DNA which resulted from its covalent modification. Use of this assay is illustrated in studies which screened and compared the activities of several recently synthesized psoralen derivatives. We describe two new compounds. β‐(8‐psoralenoxy)‐ethanol and β‐(8‐psoralenoxy)ethylamine that are significantly more active than either 8‐methoxypsoralen or trioxsalen in the biological assay