A cell-free system for functional centromere and kinetochore assembly

  title={A cell-free system for functional centromere and kinetochore assembly},
  author={Annika Guse and Colin J. Fuller and Aaron F Straight},
  journal={Nature Protocols},
This protocol describes a cell-free system for studying vertebrate centromere and kinetochore formation. We reconstitute tandem arrays of centromere protein A (CENP-A) nucleosomes as a substrate for centromere and kinetochore assembly. These chromatin substrates are immobilized on magnetic beads and then incubated in Xenopus egg extracts that provide a source for centromere and kinetochore proteins and that can be cycled between mitotic and interphase cell cycle states. This cell-free system… CONTINUE READING
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