Table 3 Effect of terminator elements ahead of the I-SceI cleavage site on introduction of sequence encoding a C-terminal 3XFLAG tag on GapA in E. coli K-12 MG1655 and S. enterica Typhimurium SL1344
E. coliK-12 MG1655 | S. entericaTyphimurium SL1344 | |||||
|---|---|---|---|---|---|---|
Number of terminators in front of the I-SceI recognition site | 0 | 1 | 2 | 0 | 1 | 2 |
Fraction of colonies that lost the selection marker after double-strand cleavage of the mutation cassette by I-SceI | 0/100 | 2/40 | 5/11 | 0/100 | 1/40 | 4/7 |
Fraction of colonies that lost the selection marker for which a fragment of the correct size was amplified by colony PCR using primers flanking the gene of interest | NA* | 1/2 | 1/2 | NA* | 1/1 | 2/2 |
Fraction of colonies for which the correct PCR product was obtained that contained the desired mutation | NA* | 1/1 | 1/1 | NA* | 1/1 | 2/2 |