Journal: Molecular Biology and Evolution

Article Title: Adaptive Capacity of a DNA Polymerase Clamp-loader ATPase Complex

doi: 10.1093/molbev/msae013

Figure Lengend Snippet: A phage-propagation assay to measure the fitness of clamp-loader variants in a T4 bacteriophage. a) A schematic diagram of the clamp-loading cycle, from left to right, showing key steps of loading the sliding clamp around a primer-templated DNA. b) A schematic of the phage-propagation assay in which T4 del infects host cells, each of which carries a plasmid-borne copy of variant genes of the T4 clamp loader. Upon infection, the variant genes recombine into the genome of T4 del , and this genome is replicated using the proteins encoded by the variant genes. The number of copies of the recombinant genomes (and the recombinant phage) is proportional to the activity of the clamp-loader variant in the host cell. Inset: A method to generate T4 del from the wild-type T4 bacteriophage using CRISPR-based genome engineering. c) A schematic diagram showing the construction of chimeric clamp-loader variants. d) The relative fitness of the chimeric clamp-loader variants from the phage-propagation assay.

Article Snippet: Recombinant DNA reagent , CRISPR plasmid , Previous work ( ) , ... , https://benchling.com/s/seq-abaKV7JgTgAghRyR9ZIY.

Techniques: Plasmid Preparation, Variant Assay, Infection, Recombinant, Activity Assay, CRISPR