Effect of growth rate on stability and gene expression of a recombinant plasmid during continuous culture ofEscherichia coli in a non-selective medium

1992 ◽  
Vol 14 (8) ◽  
pp. 643-648 ◽  
Author(s):  
Nabil Nancib ◽  
Joseph Boudrant
1961 ◽  
Vol 7 (4) ◽  
pp. 491-505 ◽  
Author(s):  
Andreas H. W. Hauschild ◽  
Hilliard Pivnick

An apparatus is described for the continuous growth of bacteria. Brucella abortus S.19 has been grown in continuous culture for periods up to 3 weeks with populations up to 2 × 1011viable cells per ml and without the establishment of nonsmooth variants.Concentrations between 3 × 109and 2 × 1011cells per ml could be maintained as a function of the dilution rate without the requirement of a known limiting factor in the medium. In a series of steady-state conditions, the specific growth rate increased steadily up to 0.28 hour−1with decreasing population levels.Incidence of mutants was governed by the dilution rate and could also be reduced by various chelating substances.In continuous growth combined with continuous dialysis, population levels were approximately twice those obtained in continuous growth without dialysis. The effect of dialysis appears to be the continuous removal of growth-limiting metabolic products.


1999 ◽  
Vol 19 (1) ◽  
pp. 899-908 ◽  
Author(s):  
Perry Kannan ◽  
Michael A. Tainsky

ABSTRACT ras oncogene-transformed PA-1 human teratocarcinoma cells have abundant AP-2 mRNA but, paradoxically, little AP-2 transcriptional activity. We have previously shown that overexpression of AP-2 in nontumorigenic variants of PA-1 cells results in inhibition of AP-2 activity and induction of tumorigenicity similar to that caused by ras transformation of PA-1 cells. Evidence indicated the existence of a novel mechanism of inhibition of AP-2 activity involving sequestering of transcriptional coactivators. In this study, we found that PC4 is a positive coactivator of AP-2 and can restore AP-2 activity in ras-transformed PA-1 cells. Relative to vector-transfected ras cell lines,ras cell lines stably transfected with and expressing the PC4 cDNA have a diminished growth rate and exhibit a loss of anchorage-independent growth, and they are unable to induce the formation of tumors in nude mice. These data suggest that a transcriptional coactivator, like a tumor suppressor, can have a growth-suppressive effect on cells. Our experiments are the first to show that ras oncogenes and oncogenic transcription factors can induce transformation through effects on the transcription machinery rather than through specific programs of gene expression.


2004 ◽  
Vol 70 (5) ◽  
pp. 2928-2934 ◽  
Author(s):  
Maria Lövenklev ◽  
Ingrid Artin ◽  
Oskar Hagberg ◽  
Elisabeth Borch ◽  
Elisabet Holst ◽  
...  

ABSTRACT The effects of carbon dioxide, sodium chloride, and sodium nitrite on type B botulinum neurotoxin (BoNT/B) gene (cntB) expression in nonproteolytic Clostridium botulinum were investigated in a tryptone-peptone-yeast extract (TPY) medium. Various concentrations of these selected food preservatives were studied by using a complete factorial design in order to quantitatively study interaction effects, as well as main effects, on the following responses: lag phase duration (LPD), growth rate, relative cntB expression, and extracellular BoNT/B production. Multiple linear regression was used to set up six statistical models to quantify and predict these responses. All combinations of NaCl and NaNO2 in the growth medium resulted in a prolonged lag phase duration and in a reduction in the specific growth rate. In contrast, the relative BoNT/B gene expression was unchanged, as determined by the cntB-specific quantitative reverse transcription-PCR method. This was confirmed when we measured the extracellular BoNT/B concentration by an enzyme-linked immunosorbent assay. CO2 was found to have a major effect on gene expression when the cntB mRNA levels were monitored in the mid-exponential, late exponential, and late stationary growth phases. The expression of cntB relative to the expression of the 16S rRNA gene was stimulated by an elevated CO2 concentration; the cntB mRNA level was fivefold greater in a 70% CO2 atmosphere than in a 10% CO2 atmosphere. These findings were also confirmed when we analyzed the extracellular BoNT/B concentration; we found that the concentrations were 27 ng · ml−1 · unit of optical density−1 in the 10% CO2 atmosphere and 126 ng · ml−1 · unit of optical density−1 in the 70% CO2 atmosphere.


2015 ◽  
Vol 17 (4) ◽  
pp. 785-790 ◽  
Author(s):  
Fuentes-Silva Carlos ◽  
G.M. Soto-Zarazúa ◽  
I. Torres-Pacheco ◽  
R.G. Guevara-González ◽  
J.F. García-Trejo ◽  
...  

Animals ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2585
Author(s):  
Fang Jiang ◽  
Yan Lin ◽  
Linghong Miao ◽  
Jingyuan Hao

The ability of bamboo charcoal to reduce the negative effects of high dietary selenium (Se) concentrations was assessed by feeding juvenile blunt snout bream (Megalobrama amblycephala) one of five Se-rich diets (1.5 mg/kg Se; 36% protein, 8.7% lipid) containing graded levels (0–4 g/kg) of bamboo charcoal powder for eight weeks. There were four tanks (350 L) of fish (initial weight 16.0 ± 0.5 g) for each treatment, and the fish were fed to satiation four times each day. At the end of the feeding trial, all of the fish from each tank were weighed to calculate the growth performance. Blood samples were firstly obtained to collect plasma for the biochemical indexes determination. Liver tissues were then collected to determine the antioxidant enzyme activities and gene expression. Dorsal muscles were also collected to determine the nutrient composition. The results show that when the bamboo charcoal content in the Se-rich feed ranged between 0 and 3 g/kg, the weight growth rate (WGR) and specific growth rate (SGR) values increased with the higher dietary bamboo charcoal content, and the maximum WGR and SGR values were achieved when the bamboo charcoal content in the Se-rich feed was 2–3 g/kg (p < 0.05). The Se content in muscle tissues decreased significantly with the increased bamboo charcoal content (p < 0.05) in the Se-rich feed, which ranged from 0 to 4 g/kg. When the bamboo charcoal content in the Se-rich feed was 2–3 g/kg, the levels of glucose (GLU) and albumin (ALB) in plasma reached a maximum (p < 0.05), whereas the level of alkaline phosphatase (ALP) reached a minimum (p < 0.05). Additionally, the activities of catalase (CAT), total superoxide dismutase (T-SOD), total antioxidative capacity (T-AOC), and glutathione peroxidase (GSH-Px) were significantly enhanced (p < 0.05) when the bamboo charcoal content was 3 g/kg. In contrast, the malondialdehyde (MDA) level increased sharply when the bamboo charcoal content in the Se-rich feed was 1 g/kg, compared to the control group and the groups supplemented with 2–3 g/kg bamboo charcoal (p < 0.05). Regarding mRNA-level gene expression, the results show that dietary supplementation with 0 to 3 g/kg of bamboo charcoal increased the expression of keap1 and nrf2, whereas nfkb expression was inhibited (p < 0.05). The mRNA expression of the antioxidant enzymes cat, gpx, and mn-sod was consistently enhanced in the group fed with the 3 g/kg bamboo charcoal diet (p < 0.05). The expression of the pro-inflammatory cytokines tnfα and tgfβ was inhibited in the groups supplemented with 2–3 g/kg bamboo charcoal, whereas the expression of anti-inflammatory cytokines (il10) increased in the bamboo charcoal supplementation groups compared to the control group (p < 0.05). Generally, supplementation with 2–3 g/kg of bamboo charcoal in Se-rich feed improved the growth performance, physiological status, and antioxidant enzyme activities of blunt snout bream. Moreover, bamboo charcoal supplementation in Se-rich diets stimulated the antioxidant system and inhibited the inflammatory response by activating Nrf2-Keap1 and suppressing NF-κB.


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