These plasmids are employed to delete genes commonly used in selection after transformation of Saccharomyces cerevisiae strains. Complete deletion or nearly complete deletion of the gene reduces background that could result from marker locus conversion between commonly transformed pRS plasmids and residual gene sequences in the yeast host genome. Each delta0 deleter plasmid completely deletes LEU2, LYS2, MET 15, TRP1 or URA3. For ADE2, use of pdeltaADE2 deletes all but 6 C-terminal amino acid codons. By means of a one-(URA3) or two-step (other markers) transformation and selection process, sequences in common with the plasmid-contained marker genes are completely or almost completely removed from the strain. The resultant strain will exhibit no background after transformation due to marker locus conversion. Deletion of LEU2, LYS2, MET15, TRP1 or ADE2 requires a ura3 mutation to be present in the starting strain, as plasmid integration/excision requires sequential Ura+/FoaR selections. If your strain is not ura3, first use pJL164 to delete the URA3 gene.

 

Marker	Designation	ATCC number	Depositor	References
ADE2	pdeltaADE2	99604	J. Boeke	RF31850, RF4356
LEU2	pAD1	87468	J. Boeke	RF32075
LYS2	pAD2	87469	J. Boeke	RF32075
MET17	pAD4	87470	J. Boeke	RF32075
TRP1	pNKY1009	87624	E. Alani	RF4356
URA3	pJL164	87471	J. Boeke	RF32075

References