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Table 1 Efficiencies of pRKaraRed-mediated recombination under different conditions

From: Scarless and sequential gene modification in Pseudomonas using PCR product flanked by short homology regions

Conditions Positive colonies/Growing colonies (%)a Overall efficiency (%)
  Replacement with marker genesb Deletion of marker genesc  
A. L-arabinose concentration
0.05% 10/19 (53%) 9/20 (45%) 24%
0.1% 31/43 (72%) 17/20 (85%) 61%
0.2% 67/68 (99%) 20/20 (100%) 99%
0.4% 62/63 (98%) 20/20 (100%) 98%
0.8% 70/73 (96%) 20/20 (100%) 96%
1.0% 59/61 (97%) 19/20 (95%) 92%
B. Length of homology regions
50 bp 66/67 (99%) 20/20 (100%) 99%
60 bp 72/73 (99%) 20/20 (100%) 99%
100 bp 79/80 (99%) 20/20 (100%) 99%
C. Induction time
1 hours 33/39 (85%) 17/20 (85%) 72%
3 hours 63/64 (98%) 20/20 (100%) 98%
6 hours 56/57 (98%) 20/20 (100%) 98%
12 hours 48/49 (98%) 19/20 (95%) 93%
  1. phzS gene was used as target. Conditions: A, 50 bp homology region, induction of cells with different concentration of L-arabinose during 3 hours; B, different lengths of homology regions, induction of cells with 0.2% L-arabinose during 3 hours; C, 50 bp homology region, induction of cells with 0.2% L-arabinose during different time.
  2. a. Determined by PCR amplification and DNA sequencing
  3. b. Screening of CarbRSucS colonies
  4. c. Screening of CarbSSucR colonies