Table 2 Efficiencies of pRKaraRed-mediated scarless modification to different targets
Target | Size (bp) | Positive colonies/Growing colonies (%)a | Overall efficiency (%) | |
|---|---|---|---|---|
Replacement using sacB-bla cassetteb | Deletion of sacB-bla cassettec | |||
A. Deletion of genes | ||||
rsm A | 186 | 43/44 (98%) | 19/20 (95%) | 93% |
las I | 606 | 53/54 (98%) | 20/20 (100%) | 98% |
gac A | 645 | 49/50 (98%) | 18/20 (90%) | 88% |
qsc R | 714 | 36/37 (97%) | 19/20 (95%) | 92% |
las R | 720 | 56/57(98%) | 20/20 (100%) | 98% |
rhl R | 762 | 59/61(97%) | 20/20 (100%) | 97% |
phz M | 1005 | 65/68 (96%) | 19/20 (95%) | 91% |
rpo S | 1005 | 46/47 (98%) | 20/20 (100%) | 98% |
phz S | 1209 | 70/72 (97%) | 20/20 (100%) | 97% |
phz H | 1833 | 68/69 (99%) | 19/20 (95%) | 89% |
rpo D | 1854 | 52/54 (96%) | 20/20 (100%) | 96% |
pts P | 2280 | 78/80 (98%) | 19/20 (95%) | 93% |
B. Single-point mutation | ||||
phz S | 1 | 24/26 (94%) | 19/20 (95%) | 89% |
(A761T) | ||||
C. Deletion of operons | ||||
phz A1-G1 | 6267 | 47/50 (94%) | 19/20 (95%) | 89% |
phz A2-G2 | 6273 | 61/63 (97%) | 20/20 (100%) | 97% |