a strategy for gene replacements

Susan L Forsburg susan_forsburg at qm.salk.edu
Wed Mar 16 16:16:11 EST 1994


In article <ghJaBRK00iV803_n8=@andrew.cmu.edu> Morris F. Manolson,
mm6y+ at andrew.cmu.edu writes:
>and if for any reason you can not use URA3, you can always use
LYS2.  
>You can select against LYS2 by growing the yeast on plates
containing
>DL-alpha-aminoadipate.  There are similar schemes for the CAN1 and
CYH2
>genes.  ..........  hope this helps, Morrie


The lys trick doesnt work in pombe :-(
Apparently wildtype pombe are quite happy on aminoadipate 
lys1+ encodes the homologue of LYS2
The S. cerevisiae CAN1 gene can be used with a pombe can1-1 marker,
in the same way as in cerevisiae (Ekwall and Ruusala, NAR 19:1150,
1991).  I have used this CAN system for a test of plasmid shuffling,
with success.

susan

------------------------------
Susan L Forsburg
Molecular Biology and Virology Laboratory
The Salk Institute for Biological Studies
email:  susan_forsburg at qm.salk.edu
        formerly forsburg at molbiol.ox.ac.uk



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