Construction of Compatible Wide-Host-Range Shuttle Vectors for Lactic Acid Bacteria and Escherichia coli

A new collection of shuttle cloning vectors has been constructed that can be used in a broad host range, because they carry replication origins which are functional in Escherichia coli (p15A, pWV01, ColE1), Lactococcus lactis, lactobacilli, and Bacillus subtilis (pAMβ1, pWV01). These plasmids contai...

Full description

Saved in:
Bibliographic Details
Published inPlasmid Vol. 46; no. 2; pp. 106 - 116
Main Authors Perez-Arellano, I, Zuniga, M, Perez-Martinez, G
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.09.2001
Subjects
Bacillus subtilis
Bacillus subtilis - genetics
Chromosome Segregation - genetics
compatible plasmids
DNA Replication - genetics
Electrophoresis, Agar Gel
Electroporation
erythromycin transacetylase
Escherichia coli
Escherichia coli - genetics
Genetic Vectors - biosynthesis
Genetic Vectors - genetics
Kinetics
Lactobacillus
Lactobacillus - genetics
Lactobacillus casei
Lactococcus casei
Lactococcus lactis
Lactococcus lactis - genetics
quantitative PCR
Replication Origin - genetics
replication origins
Restriction Mapping
shuttle cloning vectors
Species Specificity
Transformation, Bacterial - genetics
Lactobacillus casei
compatible plasmids
quantitative PCR
shuttle cloning vectors
Online AccessGet full text

Cover

More Information
Summary:A new collection of shuttle cloning vectors has been constructed that can be used in a broad host range, because they carry replication origins which are functional in Escherichia coli (p15A, pWV01, ColE1), Lactococcus lactis, lactobacilli, and Bacillus subtilis (pAMβ1, pWV01). These plasmids contain the lacZ-T1T2 cassette from pJDC9, which allows the X-gal selection and cloning of DNA fragments that could cause plasmid instability in E. coli. In addition, they have been proved to be structurally and segregationally stable in Lactobacillus casei, in which their copy number has been determined by real-time quantitative PCR. Furthermore, the antibiotic resistance markers (β-lactamase, chloramphenicol acetyl transferase, and erythromycin transacetylase) and the theta and rolling circle replicating origins have been combined to obtain this set of compatible plasmids (pIA family) that can be cotransformed, both in lactic acid bacteria and in E. coli.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
content type line 23
ObjectType-Article-1
ObjectType-Feature-2
ISSN:0147-619X
1095-9890
DOI:10.1006/plas.2001.1531