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Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM
The slpB gene of Lactobacillus acidophilus NCFM, which differs from the slpA gene and is silent under normal conditions, was successfully amplified and ligated to the corresponding available sites on a recombinant pET-28a vector. Then the pET-28a-slpB vector was transformed into Escherichia coli DH...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6015585/ https://www.ncbi.nlm.nih.gov/pubmed/29936673 http://dx.doi.org/10.1186/s13568-018-0631-2 |
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author | Guo, Yuxing Li, Xiangyue Yang, Yao Wu, Zhen Zeng, Xiaoqun Nadari, Fawze Pan, Daodong |
author_facet | Guo, Yuxing Li, Xiangyue Yang, Yao Wu, Zhen Zeng, Xiaoqun Nadari, Fawze Pan, Daodong |
author_sort | Guo, Yuxing |
collection | PubMed |
description | The slpB gene of Lactobacillus acidophilus NCFM, which differs from the slpA gene and is silent under normal conditions, was successfully amplified and ligated to the corresponding available sites on a recombinant pET-28a vector. Then the pET-28a-slpB vector was transformed into Escherichia coli DH (DE3) and the fusion His-slpB protein was expressed by induction with 1 mM IPTG for 14 h at 37 °C. The resulting His-slpB protein (S(B)) had a relative molecular weight of 48 kDa. It was purified using a Ni-NTA column and was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot contrastive analysis. The slpA protein (S(A)) from L. acidophilus NCFM was extracted and purified. It had a relative molecular weight of 46 kDa. Circular dichroism measurements suggested that the two S-layer proteins had a high β-sheet content and a low α-helix structure content. In an adhesion experiment, S(A) displayed higher adhesive capability towards Caco-2 cells than did S(B). The results suggest that these two S-layer proteins could have biotechnological applications. |
format | Online Article Text |
id | pubmed-6015585 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-60155852018-07-06 Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM Guo, Yuxing Li, Xiangyue Yang, Yao Wu, Zhen Zeng, Xiaoqun Nadari, Fawze Pan, Daodong AMB Express Original Article The slpB gene of Lactobacillus acidophilus NCFM, which differs from the slpA gene and is silent under normal conditions, was successfully amplified and ligated to the corresponding available sites on a recombinant pET-28a vector. Then the pET-28a-slpB vector was transformed into Escherichia coli DH (DE3) and the fusion His-slpB protein was expressed by induction with 1 mM IPTG for 14 h at 37 °C. The resulting His-slpB protein (S(B)) had a relative molecular weight of 48 kDa. It was purified using a Ni-NTA column and was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot contrastive analysis. The slpA protein (S(A)) from L. acidophilus NCFM was extracted and purified. It had a relative molecular weight of 46 kDa. Circular dichroism measurements suggested that the two S-layer proteins had a high β-sheet content and a low α-helix structure content. In an adhesion experiment, S(A) displayed higher adhesive capability towards Caco-2 cells than did S(B). The results suggest that these two S-layer proteins could have biotechnological applications. Springer Berlin Heidelberg 2018-06-23 /pmc/articles/PMC6015585/ /pubmed/29936673 http://dx.doi.org/10.1186/s13568-018-0631-2 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Article Guo, Yuxing Li, Xiangyue Yang, Yao Wu, Zhen Zeng, Xiaoqun Nadari, Fawze Pan, Daodong Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM |
title | Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM |
title_full | Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM |
title_fullStr | Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM |
title_full_unstemmed | Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM |
title_short | Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM |
title_sort | molecular cloning, expression and adhesion analysis of silent slpb of lactobacillus acidophilus ncfm |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6015585/ https://www.ncbi.nlm.nih.gov/pubmed/29936673 http://dx.doi.org/10.1186/s13568-018-0631-2 |
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