These two authors contributed equally to this study.
Unique secreted–surface protein complex of Lactobacillus rhamnosus, identified by phage display
Article first published online: 11 DEC 2012
© 2012 The Authors. MicrobiologyOpen published by Blackwell Publishing Ltd.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Volume 2, Issue 1, pages 1–17, February 2013
How to Cite
MicrobiologyOpen 2013; 2(1): 1–17
- Issue published online: 13 FEB 2013
- Article first published online: 11 DEC 2012
- Manuscript Accepted: 25 OCT 2012
- Manuscript Received: 17 OCT 2012
- Massey University
- Palmerston North Medical Research Foundation
- New Zealand Foundation for Research and Technology. Grant Number: C03X0701
- Fonterra Co-operative Group Ltd.
- Ministry of Science and Innovation
Figure S1. The plasmid profiles over three rounds of affinity-screening (panning) of the HN001 phage display library using HN001 as bait. Lanes: pYW01, replicative form (covalently closed circular double-stranded DNA; cccDNA) of the phagemid vector pYW01 used to construct the phage display library; Round I; Round II; Round III; purified pools of the recombinant library (cccDNA) after indicated rounds of panning. Three black arrows indicate the DNA bands of enriched phagemids.
Figure S2. Construction of the HN001 ΔspcA mutant. (A) Vector used for construction of double cross-over mutant. (B) Map of the spcA deletion mutant and the wild-type spcA locus in HN001. (C) Confirmation Southern blotting using upstream and downstream probes (indicated in B).
Figure S3. The plasmid profiles over four rounds of affinity-screening (panning) of the HN001 phage display library using purified MBP-SpcA fusion protein as bait. The panning was carried out over four rounds of affinity selection and amplification (I–IV) to enrich the specific binders. From the second to the fourth rounds of panning, MBP and BSA were also included as control baits. pYW01 represents the phagemid used to construct the phage display library.
Table S1. Bacterial strains.
Table S2. Plasmids and phage.
Table S3. Oligonucleotides.
Data S1. Experimental procedures.
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