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LIQUID‐HOLDING EFFECTS IN U.V.‐IRRADIATED PHAGE T3 *
Author(s) -
MARTIGI S.,
HARM W.
Publication year - 1972
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.1751-1097.1972.tb06274.x
Subject(s) - chloramphenicol , irradiation , dna , bacteriophage , titer , chemistry , bacteria , strain (injury) , lambda phage , microbiology and biotechnology , biophysics , biology , antibiotics , escherichia coli , biochemistry , genetics , antibody , physics , anatomy , gene , nuclear physics
Abstract— Holding complexes of u.v.‐irradiated (254 nm) T3 phage in E. coli B/r cells for several hours at 37°C in buffer, or broth with chloramphenicol, affects the phage survival in at least two different ways: (1) by enhancing excision repair, resulting under certain conditions in liquid‐holding recovery (LHR), and (2) by destroying the phage ( holding inactivation ). LHR is most apparent in buffer containing 20 μg ml ‐1 chloramphenicol (CAP). It is expressed by as much as a 10–fold increase in the fraction of complexes that display host‐cell reactivation (resulting from excision repair), but the percentage of u.v. lesions repaired within repair‐proficient complexes is slightly decreased. LHR is not observed if T3 infects the repair‐deficient strain B s‐1 . Holding inactivation is readily observed with unirradiated phage complexes in broth containing CAP. The response of irradiated‐phage complexes to liquid‐holding conditions is more complex: holding inactivation is less effective for irradiated than for unirradiated phage DNA (i.e. the irradiated DNA is to some extent ‘protected’), and processes leading to LHR are superimposed. Thus under certain holding conditions one observes the paradoxical phenomenon that the viable titer of irradiated phage is several times higher than that of unirradiated phage. The nature of holding inactivation is not known, nor is the mechanism by which irradiated DNA is partially protected against it. Holding inactivation does not require protein synthesis; it is rather enhanced at high CAP concentration and seems to be favored by otherwise active cell metabolism. At high CAP concentrations (200–400 μg ml ‐1 , as compared to 20 μg ml ‐1 ) irradiated‐phage complexes show neither LHR nor protection against holding inactivation. Likewise they fail to undergo some step by which the phage DNA becomes insensitive to repair inhibition by caffeine.