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- Title
Cloning and detection of serpin-like protein encoding gene in Bifidobacterium longum strains.
- Authors
Nissen, Lorenzo; Pasini, Luca; Biavati, Bruno; Sgorbati, Barbara
- Abstract
Bifidobacterium longum has immunostimulant activity that in vitro can lead to apoptotic response of Caco-2 cell line. It secretes a hypothetical eukaryotic type serpin protein, which could be involved in this kind of deletion of damaged cells. We had previously characterised a protein that has homologies with the hypothetical serpin of B. longum (DD087853). In order to create Bifidobacterium serpin transformants, a B. longum cosmid library was screened with a PCR protocol using specific primers for serpin gene. After fragment extraction, the insert named S1 was sub-cloned into pRM2, an Escherichia coli — Bifidobacterium shuttle vector, to construct pRM3. Several protocols for B. longum transformation were performed and the best efficiency was obtained using MRS medium and raffinose. Finally bacterial cell supernatants were tested in a dot-blot assay to detect antigens presence against anti-antitrypsin polyclonal antibody. Our research study is aimed to generate transformants able to over express serpin encoding gene, so that we can have the tools for a further study on bacterial apoptotic induction of Caco-2 cell line.
- Publication
Annals of Microbiology, 2008, Vol 58, Issue 1, p127
- ISSN
1590-4261
- Publication type
Article
- DOI
10.1007/BF03179456