engleski

The inactivation of free phages in UV-irradiated Escherichia coli

Background and purpose: The described method is based on the ability of bacteriophages to infect a UV irradiated host cell, but then fail to multiply inside the host. The host's capacity for phage multiplication as well as for its own reproduction was destroyed by very high doses of UV light. The aim of this work was to compare the efficiency of the new method with the commonly used method of phage inactivation with polyclonal rabbit anti-lambda serum. To prove the broad use of this method, three different types of E. coli phages were tested. Material and methods: Standard microbiological methods for determination of number of viable bacteria and number of viable phage particles were used. E. coli AB1157 and its derivatives were used in experiments with lambda and T4 phage, and E. coli JM101 in experiments with M13mp18. Anti-lambda serum was prepared by standard method of immunization of rabbits. Results: During incubation with anti- lambda serum (KD=193), up to 98-99 %of lambda free phages were inactivated. Almost the same results were obtained with our method of phage inactivation by UV irradiated cells. Once prepared, a suspension of irradiated cells, stored at 4C, could be used up to 10 days without significant change in inactivation ability. Comparable results were also obtained with T4 and M13mp18 phages. Conclusion: We could recommend this method because it is efficient, simple, fast and inexpensive. We presume it could be adapted for use in many experimental systems involving other types of viruses and host cells.

Escherichia coli ; bacteriophages: lambda ; T4 and M13 mp18 ; inactivation ; anti-lambda serum ; irradiation ; multiplication

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