Bacteriophage recombineering in the lytic state using the lambda red recombinases
Article first published online: 13 SEP 2011
© 2011 The Authors. Microbial Biotechnology © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd
Volume 5, Issue 4, pages 466–476, July 2012
How to Cite
Fehér, T., Karcagi, I., Blattner, F. R. and Pósfai, G. (2012), Bacteriophage recombineering in the lytic state using the lambda red recombinases. Microbial Biotechnology, 5: 466–476. doi: 10.1111/j.1751-7915.2011.00292.x
- Issue published online: 7 JUN 2012
- Article first published online: 13 SEP 2011
- Received 8 April, 2011; revised 15 July, 2011; accepted 18 July, 2011.
Fig. S1. Results of PCR reactions screening for ISes within the P1vir (A) and P1virΔIS (B) genomes, using the White Glove IS Detection Kit. In both panels, lanes contain primers specific for the following elements: lane 1: IS1, lane 2: IS2, lane 3: IS3 (ISEc17), lane 4: IS4, lane 5: IS5, lane 6: IS10, lane 7: IS30D, lane 8: IS150, lane 9: IS186, lane 10: IS600 (ISsd1), lane 11: IS609, lane 12: IS911, lane 13: ISEc1,3,5, lane 14: ISEc4, lane 15: RhsA,B,C, lane 16: RhsD,E. Lane 17 contained primers specific for P1 C1 gene. M: 1 kb DNA ladder (Fermentas).
Fig. S2. PCR verification of the position and orientation of IS1P1vir using primers IS1A1 and P1D.
Fig. S3. Dependence of phage titre on multiplicity of infection (moi).
Fig. S4. Comparison of plaque morphology. Left: P1vir. Right: P1virΔIS.
Table S1. Details of attempts to engineer P1virΔIS using BRED. Note that multiple batches of arabinose-induced electrocompetent MG1655/pKD46 cells were used during the optimization, which could cause inconsistencies.
Table S2. Primers used in this study.
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