The purpose of plasmid shuffle is to start with one plasmid in a cell, have two plasmids in the same cell, and eventually wind up with only the second plasmid in the cell.(see original paper)
1) At the beginning, there is a mutant strain of cells that is his- leu- and must be grown in media that contains both histadine and leucine. There are two plasmids that will be introduced into the strain as well.
2) Next, transform the bait plasmid (called pGAL4-bind) that provides the GAL4 binding domain and a his+ allele.
3) Now transform in the prey plasmid (pGAD) and select for His+ Leu+ cells.
4) In the yeast two-hybrid system, screen for cells that turn blue.
5) Now grow the cells in complete media for a few hours and then plate the cells on complete media.
6) Screen for cells that can grow in leu- media but cannot grow in his- media. These cells must contain only the pGAD plasmid:
7) Isolate the pGAD plasmid and transform into bacteria using ampicillin as the antibiotic.
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