scholarly journals Superoxide dismutase 1 (SOD1) and cadmium: a three models approach to the comprehension of its neurotoxic effects

2021 ◽  
Vol 4 (s1) ◽  
Author(s):  
Federica Bovio ◽  
Barbara Sciandrone ◽  
Chiara Urani ◽  
Paola Fusi ◽  
Matilde Forcella ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Caenorhabditis Elegans ◽  
Recombinant Protein ◽  
Neuronal Cells ◽  
Expression Level ◽  
Superoxide Dismutase 1 ◽  
Biological Models ◽  
Nematode Caenorhabditis Elegans ◽  
E Coli ◽  
Neurotoxic Effects

In three different biological models, the recombinant protein expressed in E. coli, the neuronal cells SH-SY5Y and the nematode Caenorhabditis elegans, cadmium inhibits SOD1 activity without affecting its expression level.

scholarly journals THE DNA OF CAENORHABDITIS ELEGANS

Genetics ◽  
1974 ◽  
Vol 77 (1) ◽  
pp. 95-104
Author(s):  
J E Sulston ◽  
S Brenner
Keyword(s):  
Caenorhabditis Elegans ◽  
Chemical Analysis ◽  
Dna Sequences ◽  
Base Composition ◽  
Nematode Caenorhabditis Elegans ◽  
5S Rna ◽  
Base Pairs ◽  
Small Component ◽  
E Coli ◽  
The Mean

ABSTRACT Chemical analysis and a study of renaturation kinetics show that the nematode, Caenorhabditis elegans, has a haploid DNA content of 8 x IO7 base pairs (20 times the genome of E. coli). Eighty-three percent of the DNA sequences are unique. The mean base composition is 36% GC; a small component, containing the rRNA cistrons, has a base composition of 51% GC. The haploid genome contains about 300 genes for 4s RNA, 110 for 5s RNA, and 55 for (18 + 28)S RNA.

MPA-capped CdTe quantum dots exposure causes neurotoxic effects in nematode Caenorhabditis elegans by affecting the transporters and receptors of glutamate, serotonin and dopamine at the genetic level, or by increasing ROS, or both

Nanoscale ◽  
2015 ◽  
Vol 7 (48) ◽  
pp. 20460-20473 ◽  
Author(s):  
Tianshu Wu ◽  
Keyu He ◽  
Qinglin Zhan ◽  
Shengjun Ang ◽  
Jiali Ying ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Caenorhabditis Elegans ◽  
Biomedical Applications ◽  
Biological Properties ◽  
Cdte Quantum Dots ◽  
Nematode Caenorhabditis Elegans ◽  
Neurotoxic Effects ◽  
Genetic Level

As quantum dots (QDs) are widely used in biomedical applications, the number of studies focusing on their biological properties is increasing.

scholarly journals Regulating the T7 RNA polymerase expression in E. coli BL21 (DE3) to provide more host options for recombinant protein production

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Fei Du ◽  
Yun-Qi Liu ◽  
Ying-Shuang Xu ◽  
Zi-Jia Li ◽  
Yu-Zhou Wang ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Rna Polymerase ◽  
Recombinant Proteins ◽  
Expression System ◽  
T7 Rna Polymerase ◽  
Expression Level ◽  
Foreign Protein ◽  
Atpase Subunit ◽  
E Coli ◽  
Prokaryotic Expression System

AbstractEscherichia coli is the most widely used bacterium in prokaryotic expression system for the production of recombinant proteins. In BL21 (DE3), the gene encoding the T7 RNA polymerase (T7 RNAP) is under control of the strong lacUV5 promoter (PlacUV5), which is leakier and more active than wild-type lac promoter (PlacWT) under certain growth conditions. These characteristics are not advantageous for the production of those recombinant proteins with toxic or growth-burdened. On the one hand, leakage expression of T7 RNAP leads to rapid production of target proteins under non-inducing period, which sucks resources away from cellular growth. Moreover, in non-inducing or inducing period, high expression of T7 RNAP production leads to the high-production of hard-to-express proteins, which may all lead to loss of the expression plasmid or the occurrence of mutations in the expressed gene. Therefore, more BL21 (DE3)-derived variant strains with rigorous expression and different expression level of T7 RNAP should be developed. Hence, we replaced PlacUV5 with other inducible promoters respectively, including arabinose promoter (ParaBAD), rhamnose promoter (PrhaBAD), tetracycline promoter (Ptet), in order to optimize the production of recombinant protein by regulating the transcription level and the leakage level of T7 RNAP. Compared with BL21 (DE3), the constructed engineered strains had higher sensitivity to inducers, among which rhamnose and tetracycline promoters had the lowest leakage ability. In the production of glucose dehydrogenase (GDH), a protein that causes host autolysis, the engineered strain BL21 (DE3::ara) exhibited higher biomass, cell survival rate and foreign protein expression level than that of BL21 (DE3). In addition, these engineered strains had been successfully applied to improve the production of membrane proteins, including E. coli cytosine transporter protein (CodB), the E. coli membrane protein insertase/foldase (YidC), and the E. coli F-ATPase subunit b (Ecb). The engineered strains constructed in this paper provided more host choices for the production of recombinant proteins.

scholarly journals Superoxide dismutase 1 (SOD1) and cadmium: a three models approach to the comprehension of its neurotoxic effects

2021 ◽  
Author(s):  
Federica Bovio ◽  
Barbara Sciandrone ◽  
Chiara Urani ◽  
Paola Fusi ◽  
Matilde Forcella ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Superoxide Dismutase 1 ◽  
Neurotoxic Effects

scholarly journals SPECTROPHOTOMETRIC METHOD TO ESTIMATE GLYCOGEN LEVELS IN THE MODEL NEMATODE CAENORHABDITIS ELEGANS

2020 ◽  
pp. 383-387
Author(s):  
Sidor ◽  
Andreyanov
Keyword(s):  
Caenorhabditis Elegans ◽  
Spectrophotometric Method ◽  
Cell Biology ◽  
Metabolic Regulation ◽  
Enzyme Analysis ◽  
Nematode Caenorhabditis Elegans ◽  
E Coli ◽  
C Elegans ◽  
Energy Maintenance

The model nematode Caenorhabditis elegans is widely used in studies on metabolic regulation and aging processes, in developmental and cell biology, neurobiology and genetics. Recent studies have shown the crucial role of glycogen, the glucose polymer, in stress resistance, energy maintenance and aging of organisms. Currently, the main methods for determining glycogen in the C. elegans nematode are iodine staining, enzyme analysis, and Raman microspectroscopy, described in 2019 by A. Cherkas. The proposed spectrophotometric method allows us to quantify glycogen levels, in contrast to those used in modern practice. The nematode was grown on NGM agar seeded with E. coli as a food source for 7 days in a thermostat at 20±20 C. Then, it was washed with M9 buffer and separated from bacteria by centrifugation for 10 min at 9 000 g. The samples were placed in an ice bath and nematodes were counted in each sample at x 10 microscope magnification. The glycogen concentration was determined spectrophotometrically. This value is average for different stages of the nematode development and amounts to 0.001821 ± 0.000009 μg.

scholarly journals Enhanced Production of Recombinant Mycobacterium tuberculosis Antigens in Escherichia coli by Replacement of Low-Usage Codons

2000 ◽  
Vol 68 (1) ◽  
pp. 233-238 ◽  
Author(s):  
David L. Lakey ◽  
Rama K. R. Voladri ◽  
Kathryn M. Edwards ◽  
Cynthia Hager ◽  
Buka Samten ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Superoxide Dismutase ◽  
Mycobacterium Tuberculosis ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Recombinant Antigen ◽  
E Coli ◽  
Enhanced Production ◽  
Antigen 85B

ABSTRACT A major obstacle to development of subunit vaccines and diagnostic reagents for tuberculosis is the inability to produce large quantities of these proteins. To test the hypothesis that poor expression of some mycobacterial genes in Escherichia coli is due, in part, to the presence of low-usage E. coli codons, we used site-directed mutagenesis to convert low-usage codons to high-usage codons for the same amino acid in the Mycobacterium tuberculosis genes for antigens 85A and 85B and superoxide dismutase. Replacement of five codons in the wild-type gene for antigen 85B increased recombinant protein production in E. coli 54-fold. The recombinant antigen elicited proliferation and gamma interferon production by lymphocytes from healthy tuberculin reactors and was recognized by monoclonal antibodies to native antigen 85, indicating that the recombinant antigen contained T-cell and B-cell epitopes. Northern blotting demonstrated only a 1.7- to 2.5-fold increase in antigen 85B mRNA, suggesting that the enhanced protein production was due primarily to enhanced efficiency of translation. Codon replacement in the genes encoding antigen 85A and superoxide dismutase yielded four- to sixfold increases in recombinant protein production, suggesting that this strategy may be generally applicable to overexpression of mycobacterial genes in E. coli.

scholarly journals Caenorhabditis elegans processes sensory information to choose between freeloading and self-defense strategies

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Jodie A Schiffer ◽  
Francesco A Servello ◽  
William R Heath ◽  
Francis Raj Gandhi Amrit ◽  
Stephanie V Stumbur ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Caenorhabditis Elegans ◽  
Sensory Information ◽  
Local Environment ◽  
Adaptive Strategy ◽  
Chemical Weapon ◽  
Nematode Caenorhabditis Elegans ◽  
Defense Strategies ◽  
E Coli ◽  
Self Defense

Hydrogen peroxide is the preeminent chemical weapon that organisms use for combat. Individual cells rely on conserved defenses to prevent and repair peroxide-induced damage, but whether similar defenses might be coordinated across cells in animals remains poorly understood. Here, we identify a neuronal circuit in the nematode Caenorhabditis elegans that processes information perceived by two sensory neurons to control the induction of hydrogen peroxide defenses in the organism. We found that catalases produced by Escherichia coli, the nematode’s food source, can deplete hydrogen peroxide from the local environment and thereby protect the nematodes. In the presence of E. coli, the nematode’s neurons signal via TGFβ-insulin/IGF1 relay to target tissues to repress expression of catalases and other hydrogen peroxide defenses. This adaptive strategy is the first example of a multicellular organism modulating its defenses when it expects to freeload from the protection provided by molecularly orthologous defenses from another species.

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