Natural transformation of Vibrio fischeri requires tfoX and tfoY
Article first published online: 1 JUN 2010
© 2010 Society for Applied Microbiology and Blackwell Publishing Ltd
Special Issue: Symbiosis. Editors: Professors Paola Bonfante, Karen Visick, and Moriya Ohkuma
Volume 12, Issue 8, pages 2302–2311, August 2010
How to Cite
Pollack-Berti, A., Wollenberg, M. S. and Ruby, E. G. (2010), Natural transformation of Vibrio fischeri requires tfoX and tfoY. Environmental Microbiology, 12: 2302–2311. doi: 10.1111/j.1462-2920.2010.02250.x
- Issue published online: 4 AUG 2010
- Article first published online: 1 JUN 2010
- Received 26 January, 2010; accepted 5 April, 2010.
Fig. S1. PCR screen confirming ainS gene replacement. Following transformation of ΔainS chromosomal DNA into wild-type recipient cells, 10 putative transformants were analysed by PCR for evidence of gene replacement of intact ainS with ΔainS::CmR. The primers used were ainScheckRev1 and camRcheckRev1 (Table S2), which will amplify a product only if the CmR cassette is within ainS. Lanes 1, 12 and 13: ladder (1KB DNA Ladder, Promega). Lanes 2–11, 14 and 15: PCR fragments. ES114 (wild-type) (lane 2), CL21 (ΔainS) (lane 3), 10 putative transformants (lanes 4–11, 14 and 15). The predicted position (∼1500 bp) of the PCR product containing an insertion of the CmR cassette into ainS is indicated by an arrow.
Fig. S2. Luminescence screen confirming ainS gene replacement. Following transformation of ΔainS chromosomal DNA into wild-type recipient cells, the luminescence of ten transformants (1–10) was measured to confirm gene replacement of the intact ainS (bright in culture) with ΔainS::CmR (dark in culture) phenotype. The luminescence of the transformants was compared with ES114 (wild-type) and CL21 (ΔainS) over a range of OD measurements.
Fig. S3. PCR analysis confirming the tfoXVF internal deletion. The replacement of the intact V. fischeri tfoX gene with ΔtfoX was confirmed by PCR using primers tfoXconfirmFor4 and tfoXconfirmRev1 (Table S2). The intact tfoXVF gene yields a ∼3400 bp PCR product; ΔtfoX yields a ∼2900 bp PCR product. 1 kb DNA Ladder, Promega (lane 1); AGP200 (ΔtfoX) (lane 2); ES114 (wild-type) (lane 3).
Table S1. Loci used for phylogenetic reconstruction.
Table S2. Oligonucleotides used in this study.
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