SacB selections

. .
Thu Jan 18 15:41:07 EST 1996


We are trying to do allele replacements using a ts replication vector
with sacB counter selection in Serratia marcescens. We select plasmid
integrants with chloramphenicol at 43oC then pop plasmid out by growing
at 30 and select for plasmid loss on sucrose because plasmid carries
sacB. The selection doesn;t seem to work in Serratia, in fact the cells
are now able to grow on sucrose as the sole carbon source when carrying
sacB (although not in the absence of the plasmid). Can any one explain
to me what might be happening and / or suggest any way to make the sacB
selection work.

Thanks


Michael Benedik               benedik at uh.edu
Biochemical Sciences
University of Houston
Houston, TX 77204-5934



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