08 - MIC170 Lecture 8(Thursday 4/23 Introducing plasmid DNA...

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MIC170 – 4/23/09 1 Lecture 8 (Thursday, 4/23) Introducing plasmid DNA into yeast cells A. Yeast- E. coli shuttle vector = cloning vector that can replicate both in yeast and E. coli (so that plasmid can be efficiently amplified in E. coli to obtain enough amounts for experiments) Any DNA can be cloned into the Yeast-E. coli shuttle vector expression of foreign gene in yeast 1. Multi-copy vector (Fig. 4.32 from the textbook) five essential elements of multi-copy vector 1. ARS = replication origin in yeast 2. ori = replication origin in E. coli 3. Selectable marker in E. coli (eg. ampicillin resistance gene) 4. Selectable marker in Yeast e.g. LEU2 gene for leu2 mutant host strains in minimal medium (without leucine) 5. Cloning site Multi-copy plasmid often fails to be transmitted to daughter cell (Fig. 5.20, Textbook)
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MIC170 – 4/23/09 2 2. CEN vector --- behaves as mini-chromosome (see the figure above) elements 1~5 + CEN sequence Low copy (1~2) but segregates properly into daughter cells
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This note was uploaded on 06/11/2009 for the course MIC 170 taught by Professor Shiozaki during the Spring '09 term at UC Davis.

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08 - MIC170 Lecture 8(Thursday 4/23 Introducing plasmid DNA...

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