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A colony color assay that measures nondisjunction and chromosome loss

A colony color assay that measures nondisjunction and chromosome loss. Hieter, Mann, Snyder, and Davis. What’s the question. Can a construct be made that will allow easy analysis of chromosome segregation?. What’s the approach?. SUP11 – ochre suppressor.

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A colony color assay that measures nondisjunction and chromosome loss

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  1. A colony color assay that measures nondisjunction and chromosome loss Hieter, Mann, Snyder, and Davis

  2. What’s the question Can a construct be made that will allow easy analysis of chromosome segregation?

  3. What’s the approach? • SUP11 – ochre suppressor. • Mutation in ade2 that is suppressed – if SUP11 is gone, cells are dark red. One copy – not so red. Two copies – closer to tan.

  4. What are the stabilities of various constructs?

  5. Does chromosome size affect stability? The more frequent occurrence of 1:0 events, probably indicates that replication is somewhat less dependable than chromosome separation

  6. What kinds of things could one do with this type of assay? • Complementation • What else?

  7. Lee and Nurse – cdc2 complementation • This paper was to show that human genes can be identified by their ability to complement yeast defects. • What would make that happen? • This is a paper someone could use for their research proposal. As are papers that are in 544 ERESERVES. • Before this, could identify related genes only by low-stringency hybridization (southern) or by cloning the gene in bacteria and looking for production of protein using antibodies.

  8. What have we covered? • Secretion- identification and ordering of mutants • GAL4 – • HSP70 • Chromosome segregation – mitosis and meiosis

  9. What things have you learned about? • Genetics, epistasis, conditional mutations, essential genes • Cloning, heterologous systems, reporter genes, controls • Analysis of multiple genes, gene replacement, use of regulatable promoters to create conditional lethals, biochemical approaches • Construction of artificial chromosomes to probe meiosis and mitosis, use of color to determine chromosome number or presence of a construct • Clone libraries • Biotechnology applications

  10. Exercise • What is a great question? • How will we approach it? • What types of biology will we use? • Mutant phenotypes • How do we determine all the proteins involved in the process? • How do we take a genomics approach?

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