A successful use of a new shuttle cloning vector pA13 for the cloning of the bacteriocins BacSJ and acidocin 8912
| Autor: | Milan Kojic, Branko Jovcic, L. Topisirovic, Ivana Strahinic, Jelena Lozo, D. Fira |
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| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Cloning
Genetics Lactococcus bacteriocin BacSJ Cloning vector food and beverages Biology Molecular cloning biology.organism_classification General Biochemistry Genetics and Molecular Biology DNA sequencing Shuttle cloning vector Open reading frame Plasmid Bacteriocin lcsh:Biology (General) plasmid pSJ2-8 General Agricultural and Biological Sciences lcsh:QH301-705.5 |
| Zdroj: | Archives of Biological Sciences, Vol 62, Iss 2, Pp 231-243 (2010) |
| ISSN: | 0354-4664 |
| Popis: | The aim of this paper was to research the molecular cloning of genes encoding the novel bacteriocin BacSJ from Lactobacillus paracasei subsp. paracasei BGSJ2-8 by using a newly constructed shuttle cloning vector pA13. A new shuttle-cloning vector, pA13, was constructed and successfully introduced into Escherichia coli, Lactobacillus and Lactococcus strains, showing a high segregational and structural stability in all three hosts. The natural plasmid pSJ2-8 from L. paracasei subsp. paracasei BGSJ2-8 was cloned in the pA13 using BamHI, obtaining the construct pB5. Sequencing and in silico analysis of the pB5 revealed 15 open reading frames (ORF). Plasmid pSJ2-8 harbors the genes encoding the production of two bacteriocins, BacSJ and acidocin 8912. The combined N-terminal amino acid sequencing of BacSJ in combination with DNA sequencing of the bacSJ2-8 gene enabled the determination of the primary structure of a bacteriocin BacSJ. The production and functional expression of BacSJ in homologous and heterologous hosts suggest that bacSJ2-8 and bacSJ2-8i together with the genes encoding the ABC transporter and accessory protein are the minimal requirement for the production of BacSJ. Biochemical and genetic analyses showed that BacSJ belongs to the class II bacteriocins. The shuttle cloning vector pA13 could be used as a tool for genetic manipulations in lactobacilli and lactococci. withdrawn; due to a printing error. Link to the Editorial Decision 10.2298/ABS1004251U |
| Databáze: | OpenAIRE |
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