scholarly journals CRISPR-Cas9: A Powerful Tool to Efficiently Engineer Saccharomyces cerevisiae

Life ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 13
Author(s):  
João Rainha ◽  
Joana L. Rodrigues ◽  
Lígia R. Rodrigues
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Yeast Genome ◽  
Specific Method ◽  
Complex Processes ◽  
Biological Studies ◽  
Precise Tool ◽  
Long Time ◽  
Gene Knockouts ◽  
Heterologous Pathway ◽  
A Current

Saccharomyces cerevisiae has been for a long time a common model for fundamental biological studies and a popular biotechnological engineering platform to produce chemicals, fuels, and pharmaceuticals due to its peculiar characteristics. Both lines of research require an effective editing of the native genetic elements or the inclusion of heterologous pathways into the yeast genome. Although S. cerevisiae is a well-known host with several molecular biology tools available, a more precise tool is still needed. The clustered, regularly interspaced, short palindromic repeats–associated Cas9 (CRISPR-Cas9) system is a current, widespread genome editing tool. The implementation of a reprogrammable, precise, and specific method, such as CRISPR-Cas9, to edit the S. cerevisiae genome has revolutionized laboratory practices. Herein, we describe and discuss some applications of the CRISPR-Cas9 system in S. cerevisiae from simple gene knockouts to more complex processes such as artificial heterologous pathway integration, transcriptional regulation, or tolerance engineering.

Taxonomic characterization and identification of Saccharomyces cerevisiae D8 for brandy production from pineapple

2018 ◽  
Vol 39 (4) ◽  
pp. 474-482
Author(s):  
Hoang Thi Le Thuong ◽  
Nguyen Quang Hao ◽  
Tran Thi Thuy
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Low Ph ◽  
Yeast Strains ◽  
Biological Studies ◽  
Taxonomic Characterization

Eight yeast strains (denoted as D1 to D8) were isolated from samples of natural fermented pineapple. Strain D8 showed highest alcoholic production at low pH and special aroma of pineapple has been chosen for further study. Taxonomic characterization of strain D8 using morphological, biochemical and molecular biological studies confirmed that strain D8  belong to Saccharomycetaceae family, Saccharomycetales order and Saccharomyces cerevisiae species. Therefore, we named this strain as Saccharomyces cerevisiae D8 for further study on Brandy production from pineapple. Citation: Hoang Thi Le Thuong, Nguyen Quang Hao, Tran Thi Thuy, 2017. Taxonomic characterization and identification of Saccharomyces cerevisiae D8 for brandy production from pineapple. Tap chi Sinh hoc, 39(4): 474- 482. DOI: 10.15625/0866-7160/v39n4.10864.*Corresponding author: [email protected] Received 5 December 2016, accepted 12 August 2017

Magnetic resonance imaging in study of lungs

2019 ◽  
pp. 10-23
Author(s):  
T. A. Akhadov ◽  
S. Yu. Guryakov ◽  
M. V. Ublinsky
Keyword(s):  
Magnetic Resonance Imaging ◽  
Magnetic Resonance ◽  
Medical Society ◽  
Resonance Imaging ◽  
Iodinated Contrast ◽  
Long Time ◽  
Women And Children ◽  
Magnetic Resonance Imaging Mri ◽  
A Current ◽  
Lung Mri

For a long time, there was a need to apply magnetic resonance imaging (MRI) technique for lung visualization in clinical practice. The development of this method is stimulated by necessity of the emergence of an alternative to computed tomography, especially when radiation and injection of iodine-containing contrast agents are contraindicated or undesirable, for example, in pregnant women and children, people with intolerance to iodinated contrast. One of the reasons why lung MRI is still rarely used is lack of elaborated standardized protocols that would be adapted to clinical needs of medical society. This publication is a current literature review on the use of MRI in lung studies.

scholarly journals Physical map of the Saccharomyces cerevisiae genome at 110-kilobase resolution.

Genetics ◽  
1991 ◽  
Vol 127 (4) ◽  
pp. 681-698 ◽  
Author(s):  
A J Link ◽  
M V Olson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Hybridization ◽  
Physical Map ◽  
Single Copy ◽  
Restriction Endonucleases ◽  
Yeast Genome ◽  
Hybridization Probes ◽  
Copy Dna ◽  
Single Copy Dna

Abstract A physical map of the Saccharomyces cerevisiae genome is presented. It was derived by mapping the sites for two restriction endonucleases, SfiI and NotI, each of which recognizes an 8-bp sequence. DNA-DNA hybridization probes for genetically mapped genes and probes that span particular SfiI and NotI sites were used to construct a map that contains 131 physical landmarks--32 chromosome ends, 61 SfiI sites and 38 NotI sites. These landmarks are distributed throughout the non-rDNA component of the yeast genome, which comprises 12.5 Mbp of DNA. The physical map suggests that those genes that can be detected and mapped by standard genetic methods are distributed rather uniformly over the full physical extent of the yeast genome. The map has immediate applications to the mapping of genes for which single-copy DNA-DNA hybridization probes are available.

scholarly journals Transcriptional regulation of ribosomal proteins during a nutritional upshift in Saccharomyces cerevisiae.

1986 ◽  
Vol 6 (7) ◽  
pp. 2429-2435 ◽  
Author(s):  
D M Donovan ◽  
N J Pearson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ribosomal Proteins ◽  
Protein Gene ◽  
Relative Rate ◽  
Ribosomal Protein Gene ◽  
Single Copy ◽  
Protein Product ◽  
Yeast Genome ◽  
Base Pairs ◽  
Beta Galactosidase

The relative rates of synthesis of Saccharomyces cerevisiae ribosomal proteins increase coordinately during a nutritional upshift. We constructed a gene fusion which contained 528 base pairs of sequence upstream from and including the TATA box of ribosomal protein gene rp55-1 (S16A-1) fused to a CYC1-lacZ fusion. This fusion was integrated in single copy at the rp55-1 locus in the yeast genome. During a nutritional upshift, in which glucose was added to cells growing in an ethanol-based medium, we found that the increase in the relative rate of synthesis of the beta-galactosidase protein product followed the same kinetics as the change in relative rates of synthesis of several ribosomal proteins measured in the same experiment. This demonstrates that the nontranscribed sequences upstream from the rp55-1 gene, which are present in the fusion, are sufficient to mediate the change in rates of synthesis characteristic of ribosomal proteins under these conditions. The results also suggest that a change in transcription rates is mainly responsible for the increase in relative rates of synthesis of ribosomal proteins during a nutritional upshift in S. cerevisiae.

scholarly journals Adaptation of central metabolite pools to variations in growth rate and cultivation conditions in Saccharomyces cerevisiae

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Kanhaiya Kumar ◽  
Vishwesh Venkatraman ◽  
Per Bruheim
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Amino Acid ◽  
Growth Rates ◽  
Pentose Phosphate Pathway ◽  
Pentose Phosphate ◽  
Relative Reduction ◽  
Central Metabolism ◽  
Cultivation Conditions ◽  
Biological Studies

Abstract Background Saccharomyces cerevisiae is a well-known popular model system for basic biological studies and serves as a host organism for the heterologous production of commercially interesting small molecules and proteins. The central metabolism is at the core to provide building blocks and energy to support growth and survival in normal situations as well as during exogenous stresses and forced heterologous protein production. Here, we present a comprehensive study of intracellular central metabolite pool profiling when growing S. cerevisiae on different carbon sources in batch cultivations and at different growth rates in nutrient-limited glucose chemostats. The latest versions of absolute quantitative mass spectrometry-based metabolite profiling methodology were applied to cover glycolytic and pentose phosphate pathway metabolites, tricarboxylic acid cycle (TCA), complete amino acid, and deoxy-/nucleoside phosphate pools. Results Glutamate, glutamine, alanine, and citrate were the four most abundant metabolites for most conditions tested. The amino acid is the dominant metabolite class even though a marked relative reduction compared to the other metabolite classes was observed for nitrogen and phosphate limited chemostats. Interestingly, glycolytic and pentose phosphate pathway (PPP) metabolites display the largest variation among the cultivation conditions while the nucleoside phosphate pools are more stable and vary within a closer concentration window. The overall trends for glucose and nitrogen-limited chemostats were increased metabolite pools with the increasing growth rate. Next, comparing the chosen chemostat reference growth rate (0.12 h−1, approximate one-fourth of maximal unlimited growth rate) illuminates an interesting pattern: almost all pools are lower in nitrogen and phosphate limited conditions compared to glucose limitation, except for the TCA metabolites citrate, isocitrate and α-ketoglutarate. Conclusions This study provides new knowledge-how the central metabolism is adapting to various cultivations conditions and growth rates which is essential for expanding our understanding of cellular metabolism and the development of improved phenotypes in metabolic engineering.

Identification and characterization of the centromere from chromosome XIV in Saccharomyces cerevisiae

1985 ◽  
Vol 5 (11) ◽  
pp. 2887-2893
Author(s):  
M Neitz ◽  
J Carbon
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Restriction Fragment ◽  
Dna Sequences ◽  
Yeast Genome ◽  
Ura3 Gene ◽  
Dna Restriction ◽  
Meiotic Analyses ◽  
The Right ◽  
In Cis

A functional centromere located on a small DNA restriction fragment from Saccharomyces cerevisiae was identified as CEN14 by integrating centromere-adjacent DNA plus the URA3 gene by homologous recombination into the yeast genome and then by localizing the URA3 gene to chromosome XIV by standard tetrad analysis. DNA sequence analysis revealed that CEN14 possesses sequences (elements I, II, and III) that are characteristic of other yeast centromeres. Mitotic and meiotic analyses indicated that the CEN14 function resides on a 259-base-pair (bp) RsaI-EcoRV restriction fragment, containing sequences that extend only 27 bp to the right of the element I to III region. In conjunction with previous findings on CEN3 and CEN11, these results indicate that the specific DNA sequences required in cis for yeast centromere function are contained within a region about 150 bp in length.

Saccharomyces cerevisiae contains two functional citrate synthase genes

1986 ◽  
Vol 6 (6) ◽  
pp. 1936-1942
Author(s):  
K S Kim ◽  
M S Rosenkrantz ◽  
L Guarente
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Citrate Synthase ◽  
Tricarboxylic Acid Cycle ◽  
Nuclear Gene ◽  
Tricarboxylic Acid ◽  
Yeast Genome ◽  
Gene Encoding ◽  
Acid Cycle ◽  
Nonfermentable Carbon Source

The tricarboxylic acid cycle occurs within the mitochondria of the yeast Saccharomyces cerevisiae. A nuclear gene encoding the tricarboxylic acid cycle enzyme citrate synthase has previously been isolated (M. Suissa, K. Suda, and G. Schatz, EMBO J. 3:1773-1781, 1984) and is referred to here as CIT1. We report here the isolation, by an immunological method, of a second nuclear gene encoding citrate synthase (CIT2). Disruption of both genes in the yeast genome was necessary to produce classical citrate synthase-deficient phenotypes: glutamate auxotrophy and poor growth on rich medium containing lactate, a nonfermentable carbon source. Therefore, the citrate synthase produced from either gene was sufficient for these metabolic roles. Transcription of both genes was maximally repressed in medium containing both glucose and glutamate. However, transcription of CIT1 but not of CIT2 was derepressed in medium containing a nonfermentable carbon source. The significance of the presence of two genes encoding citrate synthase in S. cerevisiae is discussed.

scholarly journals 1. On Galvanic Polarisation

1882 ◽  
Vol 11 ◽  
pp. 202-204
Author(s):  
Helmholtz
Keyword(s):  
Electrolytic Cell ◽  
Motive Force ◽  
Long Duration ◽  
Long Time ◽  
Electro Motive Force ◽  
A Current

In 1872 I wrote a paper on galvanic currents, which continue for a long time in an electrolytic cell, under the influence of an electro-motive force, too feeble to effect electrolytic decomposition. I tried at that time to prove that the long duration of these currents was caused by oxygen dissolved in the water of the electrolyte, combining with the hydrogen, which is carried by the electrolytic motion to the cathode. So the oxygen, which existed formerly near the surface of the cathode, is taken away, and instead of it the same amount of oxygen is liberated at the anode. This can return by diffusion to the cathode, and so the same action can go on without end. It appears as a current producing no electrolytic action. I called it “Electrolytic convection.”

scholarly journals IMPACT OF UNCERTAINTIES IN MODEL FREE PARAMETERS AND SEA LEVEL RISE ON SHORELINE CHANGES: A 20-YEAR HINDCAST AT TRUC VERT BEACH, SW FRANCE

2020 ◽  
pp. 10
Author(s):  
Maurizio D'Anna ◽  
Deborah Idier ◽  
Bruno Castelle ◽  
Goneri Le Cozannet ◽  
Jeremy Rohmer ◽  
...  
Keyword(s):  
Sea Level Rise ◽  
Sea Level ◽  
Spatial Scales ◽  
High Energy ◽  
Potential Threat ◽  
Shoreline Evolution ◽  
Complex Processes ◽  
Model Free ◽  
Long Time ◽  
Sw France

Chronic erosion of sandy coasts is a continuous potential threat for the growing coastal communities worldwide. The prediction of shoreline evolution is therefore key issue for robust decision making worldwide, especially in the context of climate change. Shorelines respond to various complex processes interacting at several temporal and spatial scales, making shoreline reconstructions and predictions challenging and uncertain, especially on long time scales (e.g. decades or century). Despite the increasing progresses in addressing uncertainties related to the physics of Sea Level Rise, very little effort is made towards understanding and reducing the uncertainties related to wave driven coastal response. To fill this gap, we analyse the uncertainties associated with long-term (2 decades) modelling of the cross-shore transport dominated high-energy sandy coast around Truc Vert beach, SW France, which has been surveyed semi-monthly over the last 12 years.Recorded Presentation from the vICCE (YouTube Link): https://youtu.be/_NBJ2v-koMs

scholarly journals Social and psychological features of inter-cultural adaptation of Russian students in different universities

2019 ◽  
Vol 14 (1) ◽  
pp. 90-98
Author(s):  
Alexey A. Kolesnikov ◽  
Darya E. Nikulina ◽  
Ilya A. Danilenko ◽  
Natalya V. Zimovets
Keyword(s):  
Bologna Process ◽  
Exchange Students ◽  
World Community ◽  
Complex Processes ◽  
Cultural Shock ◽  
Long Time ◽  
Local Residents ◽  
Adaptation Processes ◽  
Psychological Features ◽  
Foreign Universities

Modern globalisation processes, the rapid entry of Russia into the world community, have given more opportunities to interact with various ethnic groups ranging from short-term tourist and business contacts to complex processes of migration and emigration. Migrants and visitors have different goals for staying in a new country, meanwhile the researchers note a lot in common within the adaptation processes of both groups. In particular, tension, stresses and experience of cultural shock during adaptation are observed. Therefore, it is necessary to conduct psychological studies not only of migrants who come for a long time to a new country, but also of visitors, i.e., interns, students who come to study in foreign universities. Anyway, all migrants face difficulties in interacting with local residents whose behaviour cannot be predicted. The host country’s customs often seem mysterious to them, and people seem strange. Keywords: Adaptation, bologna process, ethnopsychology, exchange students.

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