The mode of replication is a major factor in segregational plasmid instability in Lactococcus lactis

The effects of the rolling-circle and theta modes of replication on the maintenance of recombinant plasmids in Lactococcus lactis were studied. Heterologous Escherichia coli or bacteriophage lambda DNA fragments of various sizes were inserted into vectors based on either the rolling-circle-type plas...

Full description

Saved in:
Bibliographic Details
Published in:Applied and Environmental Microbiology 1993-02, Vol.59 (2), p.358-364
Main Authors: Kiewiet, R. (Centre of Biological Sciences, Haren, The Netherlands), Kok, J, Seegers, J.F.M.L, Venema, G, Bron, S
Format: Article
Language:English
Subjects:
ADN
Bacteria
Biological and medical sciences
Biotechnology
Cellular biology
Deoxyribonucleic acid
DIMENSION
DIVISION CELLULAIRE
DIVISION CELULAR
DNA
Food
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Genetics and Molecular Biology
Lactococcus lactis
Methods. Procedures. Technologies
PLASMIDE
PLASMIDIOS
SEGREGACION
SEGREGATION
Vectors (cloning, transfer, expression). Insertion sequences and transposons
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The effects of the rolling-circle and theta modes of replication on the maintenance of recombinant plasmids in Lactococcus lactis were studied. Heterologous Escherichia coli or bacteriophage lambda DNA fragments of various sizes were inserted into vectors based on either the rolling-circle-type plasmid pWV01 or the theta-type plasmid pAM-beta-1. All pAM-beta-1 derivatives were stably maintained. pWV01 derivatives, however, showed size-dependent segregational instability, in particular when large DNA fragments were inserted. All recombinant pWV01 derivatives generated high-molecular-weight plasmid multimers (HMW) in amounts that were positively correlated with plasmid size and inversely correlated with the copy numbers of the monomeric plasmid forms. Formation of HMW or reductions in copy numbers were not observed with pAM-beta-1 derivatives. The results indicate that HMW formation and/or reduction in plasmid copy numbers is an important factor in the maintenance of pWV01 derivatives. It is concluded that theta-type plasmids are superior to rolling-circle-type plasmids for cloning in lactococci
ISSN:0099-2240
1098-5336
DOI:10.1128/aem.59.2.358-364.1993