Journal Article PreJuSER-57849

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Versatile Expression and Secretion Vectors for Bacillus subtilis

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2006
Springer New York, NY

Current microbiology 52, 143 - 148 () [10.1007/s00284-005-0231-7]

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Abstract: Most expression systems are based on Escherichia coli as the host strain because of the large availability of all kinds of vector plasmids. However, aside from the obvious advantages of E. coli systems, serious problems can occur during the process of heterologous gene expression and purification. Therefore, low expression rates, formation of inclusion bodies, improper protein-folding, and/or toxicity problems might enforce changing the expression host. Here we describe the construction of two new vectors, pBSMuL1 and pBSMuL2, for overexpression and secretion of heterologous proteins in Bacillus subtilis as an alternative expression host. The new plasmids combine several advantages in comparison to available Bacillus expression systems: an appropriate multiple cloning site consisting of 13 unique restriction sites, one (pBSMuL1) or two (pBSMuL2) strong constitutive promoters, a high efficient signal sequence for protein secretion, and the possibility to express proteins as His-tagged fusions for easy detection and purification. We have demonstrated the applicability of the novel vector plasmids for the production and purification of the heterologous cutinase from Fusarium solani pisi.

Keyword(s): Bacillus subtilis: metabolism (MeSH) ; Base Sequence (MeSH) ; Carboxylic Ester Hydrolases: biosynthesis (MeSH) ; Carboxylic Ester Hydrolases: genetics (MeSH) ; Fusarium: enzymology (MeSH) ; Genetic Vectors: genetics (MeSH) ; Molecular Sequence Data (MeSH) ; Plasmids (MeSH) ; Protein Engineering: methods (MeSH) ; Recombinant Proteins: biosynthesis (MeSH) ; Recombinant Proteins ; Carboxylic Ester Hydrolases ; cutinase ; J

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  1. ohne FE (ohne FE)

Appears in the scientific report 2006
Notes: Nachtrag
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 Record created 2012-11-13, last modified 2018-02-11



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