scholarly journals Enhancement of plasmid DNA transformation efficiencies in early stationary-phase yeast cell cultures

Yeast ◽  
2013 ◽  
Vol 30 (5) ◽  
pp. 191-200 ◽  
Author(s):  
Jennifer DeMars Tripp ◽  
Jennifer L. Lilley ◽  
Whitney N. Wood ◽  
L. Kevin Lewis
Keyword(s):  
Stationary Phase ◽  
Yeast Cell ◽  
Cell Cultures ◽  
Plasmid Dna ◽  
Dna Transformation ◽  
Early Stationary Phase

scholarly journals The rye Mutants Identify a Role for Ssn/Srb Proteins of the RNA Polymerase II Holoenzyme During Stationary Phase Entry in Saccharomyces cerevisiae

Genetics ◽  
2001 ◽  
Vol 157 (1) ◽  
pp. 17-26 ◽  
Author(s):  
Ya-Wen Chang ◽  
Susie C Howard ◽  
Yelena V Budovskaya ◽  
Jasper Rine ◽  
Paul K Herman
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Growth ◽  
Rna Polymerase ◽  
Stationary Phase ◽  
Yeast Cell ◽  
Rna Polymerase Ii ◽  
Transcriptional Response ◽  
Nutrient Deprivation ◽  
Ras Signaling ◽  
Rna Polymerase Ii Holoenzyme

Abstract Saccharomyces cerevisiae cells enter into a distinct resting state, known as stationary phase, in response to specific types of nutrient deprivation. We have identified a collection of mutants that exhibited a defective transcriptional response to nutrient limitation and failed to enter into a normal stationary phase. These rye mutants were isolated on the basis of defects in the regulation of YGP1 expression. In wild-type cells, YGP1 levels increased during the growth arrest caused by nutrient deprivation or inactivation of the Ras signaling pathway. In contrast, the levels of YGP1 and related genes were significantly elevated in the rye mutants during log phase growth. The rye defects were not specific to this YGP1 response as these mutants also exhibited multiple defects in stationary phase properties, including an inability to survive periods of prolonged starvation. These data indicated that the RYE genes might encode important regulators of yeast cell growth. Interestingly, three of the RYE genes encoded the Ssn/Srb proteins, Srb9p, Srb10p, and Srb11p, which are associated with the RNA polymerase II holoenzyme. Thus, the RNA polymerase II holoenzyme may be a target of the signaling pathways responsible for coordinating yeast cell growth with nutrient availability.

scholarly journals 6S RNA regulation of relA alters ppGpp levels in early stationary phase

Microbiology ◽  
2010 ◽  
Vol 156 (12) ◽  
pp. 3791-3800 ◽  
Author(s):  
Amy T. Cavanagh ◽  
Pete Chandrangsu ◽  
Karen M. Wassarman
Keyword(s):  
Amino Acid ◽  
Stationary Phase ◽  
Regulation Of Transcription ◽  
Early Stationary Phase ◽  
Rna Regulation ◽  
Global Regulators ◽  
Expression Of Genes ◽  
Non Coding Rna ◽  
6S Rna ◽  
Altered Regulation

6S RNA is a small, non-coding RNA that interacts directly with σ 70-RNA polymerase and regulates transcription at many σ 70-dependent promoters. Here, we demonstrate that 6S RNA regulates transcription of relA, which encodes a ppGpp synthase. The 6S RNA-dependent regulation of relA expression results in increased ppGpp levels during early stationary phase in cells lacking 6S RNA. These changes in ppGpp levels, although modest, are sufficient to result in altered regulation of transcription from σ 70-dependent promoters sensitive to ppGpp, including those promoting expression of genes involved in amino acid biosynthesis and rRNA. These data place 6S RNA as another player in maintaining appropriate gene expression as cells transition into stationary phase. Independent of this ppGpp-mediated 6S RNA-dependent regulation, we also demonstrate that in later stationary phase, 6S RNA continues to downregulate transcription in general, and specifically at a subset of the amino acid promoters, but through a mechanism that is independent of ppGpp and which we hypothesize is through direct regulation. In addition, 6S RNA-dependent regulation of σ S activity is not mediated through observed changes in ppGpp levels. We suggest a role for 6S RNA in modulating transcription of several global regulators directly, including relA, to downregulate expression of key pathways in response to changing environmental conditions.

scholarly journals Plasmid DNA Supercoiling and Gyrase Activity in Escherichia coli Wild-Type and rpoS Stationary-Phase Cells

2003 ◽  
Vol 185 (3) ◽  
pp. 1097-1100 ◽  
Author(s):  
Yazmid Reyes-Domínguez ◽  
Gabriel Contreras-Ferrat ◽  
Jesús Ramírez-Santos ◽  
Jorge Membrillo-Hernández ◽  
M. Carmen Gómez-Eichelmann
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Plasmid Dna ◽  
Bimodal Distribution ◽  
Dna Supercoiling ◽  
Wild Type

ABSTRACT Stationary-phase cells displayed a distribution of relaxed plasmids and had the ability to recover plasmid supercoiling as soon as nutrients became available. Preexisting gyrase molecules in these cells were responsible for this recovery. Stationary-phase rpoS cells showed a bimodal distribution of plasmids and failed to supercoil plasmids after the addition of nutrients, suggesting that rpoS plays a role in the regulation of plasmid topology during the stationary phase.

scholarly journals Chitosan Elicitation for Enhancing of Vincristine and Vinblastine Accumulation in Cell Culture of Catharanthus roseus (L.) G. Don

2018 ◽  
Vol 10 (12) ◽  
pp. 287
Author(s):  
Pawnpirun Pliankong ◽  
Padungsak Suksa-Ard ◽  
Surawit Wannakrairoj
Keyword(s):  
Molecular Weight ◽  
Cell Culture ◽  
Stationary Phase ◽  
Liquid Medium ◽  
Cell Cultures ◽  
Catharanthus Roseus ◽  
Dry Weight ◽  
Shrimp Shell ◽  
Anti Cancer ◽  
Herbal Plant

Catharanthus roseus (L.) G. Don is an important herbal plant. There are two important alkaloids, vinblastine and vincristine, use in anti-cancer drugs. In this study production of the two alkaloids was enhanced in C. roseus cell cultures, in a Murashige and Skoog (MS) liquid medium supplemented with 1.5 mg/L 2,4-D, 0.5 mg/L kinetin and 30 g/L sucrose, by adding 0, 50, 100, 250 or 500 mg/L medium molecular weight chitosan or chitosan derived from shrimp shell. After 14 days of culture, the cell suspension at stationary phase in the 100 mg/L medium molecular weight chitosan could produce the highest amounts of vinblastine and vincristine at 4.15 and 5.48 µg/mg cell dry weight, respectively. At the same time, the controls (0 mg/L chitosan) produced the two alkaloids at only 2.43 and 2.49 µg/mg cell dry weight, respectively. For chitosan from shrimp shell, it was found that 100 mg/L chitosan could lead to the highest quantity of 4.09 µg vinblastine/mg cell dry weight. The highest amount of 5.47 µg vincristine/mg cell dry weight was obtained when 250 mg/L chitosan was added.

Inducing Yeast Cell Synchrony: Dilution of Stationary-Phase Cultures

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot4176
Author(s):  
David C. Amberg ◽  
Daniel J. Burke ◽  
Jeffrey N. Strathern
Keyword(s):  
Stationary Phase ◽  
Yeast Cell ◽  
Cell Synchrony

The effect of sublethal heating on Staphylococcus aureus at different physiological ages

1974 ◽  
Vol 20 (5) ◽  
pp. 765-768 ◽  
Author(s):  
Andre Hurst ◽  
Ashton Hughes ◽  
David L. Collins-Thompson
Keyword(s):  
Staphylococcus Aureus ◽  
Stationary Phase ◽  
Heat Resistance ◽  
Growth Cycle ◽  
High Salt ◽  
Early Stationary Phase ◽  
Heat Injury ◽  
The Difference

Sublethal heat injury was measured by the difference in the numbers of colonies developing on trypticase soy agar and trypticase soy agar containing 7.5% NaCl. This difference was largest with late logarithmic and early stationary phase cells because, at this stage, cells had a greatly increased heat resistance. In contrast, the ability to form colonies on high salt agar after sublethal heating varied little during the growth cycle.

Physiological effects of plasmid DNA transformation on Azotobacter vinelandii

1986 ◽  
Vol 32 (2) ◽  
pp. 145-148 ◽  
Author(s):  
Baernard R. Glick ◽  
Heather E. Brooks ◽  
J. J. Pasternak
Keyword(s):  
Plasmid Dna ◽  
Azotobacter Vinelandii ◽  
Genetic Manipulation ◽  
A Priori ◽  
Broad Host Range ◽  
Physiological Effects ◽  
Dna Transformation ◽  
Physiological Changes ◽  
Physiological Processes ◽  
Broad Host Range Plasmid

Genetic transformation of Azotobacter vinelandii by the introduction of broad-host-range plasmid DNA (i.e., pRK2501, RSF1010, or pGSS15) causes a number of physiological changes. As shown here, the capacity for nitrogen fixation, mean cell size, and synthesis of siderophores are decreased, whereas the production of capsular slime is enhanced. These findings suggest that the presence of plasmid DNA imposes a "metabolic load" on Azotobacter vinelandii. Therefore, it cannot be assumed a priori that the introduction of plasmid DNA into Azotobacter vinelandii will not disrupt some normal physiological processes. The implications of these findings are discussed, specifically in the context of developing Azotobacter vinelandii as an effective bacterial fertilizer by genetic manipulation.

Transformation methods for halophilic archaebacteria

1989 ◽  
Vol 35 (1) ◽  
pp. 148-152 ◽  
Author(s):  
Steven W. Cline ◽  
Wan L. Lam ◽  
Robert L. Charlebois ◽  
Leonard C. Schalkwyk ◽  
W. Ford Doolittle
Keyword(s):  
Molecular Weight ◽  
Plasmid Dna ◽  
Genomic Dna ◽  
Halobacterium Halobium ◽  
Dna Transformation ◽  
Dna Transfection ◽  
Phage Dna ◽  
Halophilic Archaebacteria ◽  
Transformation Methods ◽  
Halobacterium Volcanii

We present a practical description of polyethylene glycol mediated spheroplast transformation of Halobacterium halobium and Halobacterium volcanii. This method has been applied to phage DNA transfection, plasmid DNA transformation, and transformation with linear fragments of high molecular weight genomic DNA. Efficient spheroplast regeneration allows uncomplicated recovery of transformed progeny. Transformations can be performed equally well using fresh or frozen cell preparations. These methods should find application in molecular cloning, genetic fine mapping, and strain construction.Key words: archaebacteria, Halobacterium, transformation methods, spheroplast.

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