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Title
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Flow cytometric testing of green fluorescent protein-tagged **Lactobacillus rhamnosus** GG for response to defensins
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Author
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Abstract
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| Lactobacillus rhamnosus GG is of general interest as a probiotic. Although L. rhamnosus GG is often used in clinical trials, there are few genetic tools to further determine its mode of action or to develop it as a vehicle for heterologous gene expression in therapy. Therefore, we developed a reproducible, efficient electroporation procedure for L. rhamnosus GG. The best transformation efficiency obtained was 104 transformants per μg of DNA. We validated this protocol by tagging L. rhamnosus GG with green fluorescent protein (GFP) using the nisin-controlled expression (NICE) system. Parameters for overexpression were optimized, which allowed expression of gfp in L. rhamnosus GG upon induction with nisin. The GFP+ strain can be used to monitor the survival and behavior of L. rhamnosus GG in vivo. Moreover, implementation of the NICE system as a gene expression switch in L. rhamnosus GG opens up possibilities for improving and expanding the performance of this strain. The GFP-labeled strain was used to demonstrate that L. rhamnosus GG is sensitive to human beta-defensin-2 but not to human beta-defensin-1. |
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Language
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English
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Source (journal)
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Applied and environmental microbiology. - Baltimore, Md
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Publication
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Baltimore, Md
:
2006
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ISSN
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0099-2240
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DOI
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10.1128/AEM.02605-05
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Volume/pages
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72
:7
(2006)
, p. 4923-4930
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ISI
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000238961000053
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Full text (Publisher's DOI)
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Full text (publisher's version - intranet only)
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