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Sears, Hegemann, and Heiter

Meiotic recombination and segregation of human-derived artificial chromosomes in Saccharomyces cerevisiae. Sears, Hegemann, and Heiter. What’s the question. Is it possible to make an artificial chromosome?

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Sears, Hegemann, and Heiter

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  1. Meiotic recombination and segregation of human-derived artificial chromosomes in Saccharomyces cerevisiae Sears, Hegemann, and Heiter

  2. What’s the question • Is it possible to make an artificial chromosome? • Can these be used to study nondisjunction and recombination and the relationship between these two events?

  3. What’s the approach? • Molecular biology with genetics analysis

  4. For more information, we have to go back to the first YAC paper What do we need to know to read this paper? They have already made YACs. How was this done?

  5. YAC construction • Make plasmid with telomeric region (Y’) • Add MCS • Add Alu regions for recombination sites (don’t have these sequences in yeast DNA) Couldn’t study nondisjunction of these because there are essential genes on every chromosome.

  6. What did they find? What is the problem? What is their solution? What did they find?

  7. Deleting a centromere Used a Hind III fragment

  8. What is the genetic analysis?

  9. What do we need to know? PD, NPD, TT used to determine linkage. http://dbb.urmc.rochester.edu/labs/sherman_f/yeast/7.html

  10. Now, what does the data show? High degree of linkage No double crossovers Absolute linkage, no recombination

  11. Conclusion • Can use these constructs to study recombination and linkage

  12. Using these constructs to identify non-disjunction: Work through these with drawings Homologous chromosomes Meiosis 1 Meiosis 2

  13. What would they see if there were non-disjunction?

  14. Does pairing/recombination affect nondisjunction?

  15. What was the outcome of this work? • YACs became the basis for much of the human genome sequencing effort • They were eventually retired in favor of BACs, which were easier to isolate and generally more stable.

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