TheCDC6Gene Is Required for Centromeric, Episomal, and 2-μm Plasmid Stability in the YeastSaccharomyces cerevisiae

Plasmid ◽  
1995 ◽  
Vol 34 (3) ◽  
pp. 184-197 ◽  
Author(s):  
Francesca Storici ◽  
Jacques Oberto ◽  
Carlo V. Bruschi
Keyword(s):  
Plasmid Stability ◽  
2 Μm Plasmid

Evidence for Darwinian selection of the 2-μm plasmid STB locus in Saccharomyces cerevisiae

Genome ◽  
1994 ◽  
Vol 37 (1) ◽  
pp. 12-18 ◽  
Author(s):  
G. H. Rank ◽  
W. Xiao ◽  
G. M. Arndt
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasmid Stability ◽  
Type I ◽  
Type Ii ◽  
Haploid Strain ◽  
Unique Region ◽  
Cis Acting ◽  
Plasmid Partition ◽  
Wine Strain ◽  
2 Μm Plasmid

The 2-μm plasmid of industrial and laboratory strains of Saccharomyces cerevisiae exists as two main polymorphic forms designated type I and type II. Polymorphism is restricted to the 3200-bp right unique region where types I and II show approximately 10% nucleotide divergence in trans-acting REP1 and RAF loci and 30% divergence in the cis-acting STB locus. In addition, the cis-acting STB plasmid partition locus of type II plasmids varies in sequence and copy number of a 125-bp repeat. We devised chimeric and 2-μm plasmid stability experiments to evaluate the effect of STB polymorphism on plasmid fitness in amphiploid industrial and haploid laboratory strains. Reciprocal experiments of type-II STB chimeric plasmids in type-I bakers' yeast or a type-I chimeric plasmid in type-II distillers', wine, or haploid strains showed similar partition efficiencies. However, chimeric and 2-μm plasmids carrying a 250-bp STB from a type-II haploid strain had reduced fitness in a type-II industrial wine strain. These results in conjunction with molecular analyses of 2-μm-like and 2-μm plasmids indicates the coevolution of STB with trans-acting plasmid and host-cell factors.Key words: Saccharomyces cerevisiae, 2-μm plasmid, STB adaption.

DNA insertions in the ‘silent’ regions of the 2 μm plasmid ofSaccharomyces cerevisiae influence plasmid stability

Yeast ◽  
1991 ◽  
Vol 7 (4) ◽  
pp. 347-356 ◽  
Author(s):  
J. F. M. Bijvoet ◽  
A. L. Van Der Zanden ◽  
N. Goosen ◽  
J. Brouwer ◽  
P. Van De Putte
Keyword(s):  
Plasmid Stability ◽  
2 Μm Plasmid

Overexpression of the dctA gene in Rhizobium meliloti: effect on transport of C4 dicarboxylates and symbiotic nitrogen fixation

1992 ◽  
Vol 38 (6) ◽  
pp. 555-562 ◽  
Author(s):  
Vipin Rastogi ◽  
Monika Labes ◽  
Turlough Finan ◽  
Robert Watson
Keyword(s):  
Nitrogen Fixation ◽  
Acetylene Reduction ◽  
Symbiotic Nitrogen Fixation ◽  
Plasmid Stability ◽  
Rhizobium Meliloti ◽  
Mrna Synthesis ◽  
Dicarboxylate Transport ◽  
Fusion Strain ◽  
Tryptophan Operon ◽  
Specific Mrna

Symbiotic nitrogen fixation may be limited by the transport of C4 dicarboxylates into bacteroids in the nodule for use as a carbon and energy source. In an attempt to increase dicarboxylate transport, a plasmid was constructed in which the Rhizobium meliloti structural transport gene dctA was fused to a tryptophan operon promoter from Salmonella typhimurium, trpPO. This resulted in a functional dctA gene that was no longer under the control of the dctBD regulatory genes, but the recombinant plasmid was found to be unstable in R. meliloti. To stably integrate the trpPO-dctA fusion, it was recloned into pBR325 and recombined into the R. meliloti exo megaplasmid in the dctABD region. The resultant strain showed constitutive dctA-specific mRNA synthesis which was about 5-fold higher than that found in fully induced wild-type cells. Uptake assays showed that [14C]succinate transport by the trpPO-dctA fusion strain was constitutive, and the transport rate was the same as that of induced control cells. Acetylene reduction assays indicated a significantly higher rate of nitrogen fixation in plants inoculated with the trpPO-dctA fusion strain compared with the control. Despite this apparent increase, the plants had the same top dry weights as those inoculated with control cells. Key words: acetylene reduction, genetic engineering, nodule, plasmid stability, promoter.

scholarly journals Plasmid stability analysis based on a new theoretical model employing stochastic simulations

PLoS ONE ◽  
2017 ◽  
Vol 12 (8) ◽  
pp. e0183512 ◽  
Author(s):  
Olesia Werbowy ◽  
Sławomir Werbowy ◽  
Tadeusz Kaczorowski
Keyword(s):  
Stability Analysis ◽  
Theoretical Model ◽  
Plasmid Stability ◽  
Stochastic Simulations

Linear plasmids in Klebsiella and other Enterobacteriaceae

2021 ◽  
Author(s):  
Jane Hawkey ◽  
Hugh Cottingham ◽  
Alex Tokolyi ◽  
Ryan R Wick ◽  
Louise M Judd ◽  
...  
Keyword(s):  
Plasmid Stability ◽  
Bacterial Species ◽  
Salmonella Typhi ◽  
Multidrug Resistant ◽  
Linear Plasmid ◽  
Extrachromosomal Dna ◽  
Linear Plasmids ◽  
Future Studies ◽  
And Function

Linear plasmids are extrachromosomal DNA that have been found in a small number of bacterial species. To date, the only linear plasmids described in the Enterobacteriaceae family belong to Salmonella, first found in Salmonella Typhi. Here, we describe a collection of 12 isolates of the Klebsiella pneumoniae species complex in which we identified linear plasmids. We used this collection to search public sequence databases and discovered an additional 74 linear plasmid sequences in a variety of Enterobacteriaceae species. Gene content analysis divided these plasmids into five distinct phylogroups, with very few genes shared across more than two phylogroups. The majority of linear plasmid-encoded genes are of unknown function, however each phylogroup carried its own unique toxin-antitoxin system and genes with homology to those encoding the ParAB plasmid stability system. Passage in vitro of the 12 linear plasmid-carrying Klebsiella isolates in our collection (which include representatives of all five phylogroups) indicated that these linear plasmids can be stably maintained, and our data suggest they can transmit between K. pneumoniae strains (including members of globally disseminated multidrug resistant clones) and also between diverse Enterobacteriaceae species. The linear plasmid sequences, and representative isolates harbouring them, are made available as a resource to facilitate future studies on the evolution and function of these novel plasmids.

Histone H1 expressed in Saccharomyces cerevisiae binds to chromatin and affects survival, growth, transcription, and plasmid stability but does not change nucleosomal spacing

1994 ◽  
Vol 14 (4) ◽  
pp. 2822-2835
Author(s):  
C Linder ◽  
F Thoma
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasmid Stability ◽  
Apparent Molecular Weight ◽  
Histone H1 ◽  
Expression Levels ◽  
Inhibition Of Growth ◽  
Gene Coding ◽  
Gal1 Promoter ◽  
Core Histones ◽  
Low Levels

Histone H1 is proposed to serve a structural role in nucleosomes and chromatin fibers, to affect the spacing of nucleosomes, and to act as a general repressor of transcription. To test these hypotheses, a gene coding for a sea urchin histone H1 was expressed from the inducible GAL1 promoter in Saccharomyces cerevisiae by use of a YEp vector for high expression levels (strain YCL7) and a centromere vector for low expression levels (strain YCL1). The H1 protein was identified by its inducibility in galactose, its apparent molecular weight, and its solubility in 5% perchloric acid. When YCL7 was shifted from glucose to galactose for more than 40 h to achieve maximal levels of H1, H1 could be copurified in approximately stoichiometric amounts with core histones of Nonidet P-40-washed nuclei and with soluble chromatin fractionated on sucrose gradients. While S. cerevisiae tolerated the expression of low levels of H1 in YCL1 without an obvious phenotype, the expression of high levels of H1 correlated with greatly reduced survival, inhibition of growth, and increased plasmid loss but no obvious change in the nucleosomal repeat length. After an initial induction, RNA levels for GAL1 and H1 were drastically reduced, suggesting that H1 acts by the repression of galactose-induced genes. Similar effects, but to a lower extent, were observed when the C-terminal tail of H1 was expressed.

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