Tracking bacterial DNA replication forksin vivoby pulsed field gel electrophoresis

Abstract
The location of chromosomal DNA replication forks was identified in synhronously replicating Escherichia coli cultures by pulse labeling DNA at specific times with 14C-thymidine and following incorporation of radionucleotide into genomic Not I restriction fragments. This technique could be used to characterize chromosomal DNA replication, to characterize mutations which affect this process, to identify the location of DNA replication origins and termini as well as aid in the construction of macrorestriction maps. Here, we further characterize the DNA replication mutations divE and dnaK and preliminarily characterize the genomic organization E. coli isolate 15.

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