scholarly journals Single-Step Gene Knockout of the SUC2 Gene in Saccharomyces cerevisiae : A Laboratory Exercise for Undergraduate Students †

2018 ◽  
Vol 19 (3) ◽  
Author(s):  
Jurre Hageman ◽  
Arjen M. Krikken
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Undergraduate Students ◽  
Gene Knockout ◽  
Single Step ◽  
Laboratory Exercise ◽  
Suc2 Gene

A laboratory exercise to illustrate increased salivary cortisol in response to three stressful conditions using competitive ELISA

2007 ◽  
Vol 31 (1) ◽  
pp. 110-115 ◽  
Author(s):  
Mark F. Haussmann ◽  
Carol M. Vleck ◽  
Eugenia S. Farrar
Keyword(s):  
Undergraduate Students ◽  
Salivary Cortisol ◽  
Systemic Circulation ◽  
Competitive Elisa ◽  
State University ◽  
Iowa State University ◽  
Laboratory Exercise ◽  
Comparative Endocrinology ◽  
Elisa Technique ◽  
Laboratory Tool

Perceived stress activates the hypothalamus-pituitary-adrenal axis, resulting in the release of glucocorticoids into the systemic circulation. Glucocorticoids cause the elevation of blood glucose, providing the necessary energy for the organism to cope with stress. Here, we outline a laboratory exercise that uses a competitive ELISA kit to illustrate the response of salivary cortisol concentrations to three stressful conditions. Twelve undergraduate students in the General and Comparative Endocrinology course at Iowa State University were subjected to presentation stress, fasting stress, and competition stress to determine their effect on salivary cortisol concentrations. Students had elevated salivary cortisol in response to each of these stresses compared with basal conditions. These results reiterate the importance of glucocorticoids as mediators of the stress response. This study also incorporates the use of the ELISA technique, a modern laboratory tool used to determine the amount of endogenous antigens in plasma or saliva. This laboratory exercise can easily be adapted to fit into already existing physiology and endocrinology curriculums.

scholarly journals Effect of the Deletion of Genes Related to Amino Acid Metabolism on the Production of Higher Alcohols by Saccharomyces cerevisiae

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Ya-Ping Wang ◽  
Xiao-Qing Wei ◽  
Xue-Wu Guo ◽  
Dong-Guang Xiao
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acid ◽  
Amino Acid Metabolism ◽  
Acid Metabolism ◽  
Gene Knockout ◽  
Great Influence ◽  
Regulation Mechanism ◽  
Higher Alcohols ◽  
Higher Alcohol ◽  
Alcohol Production

The higher alcohols produced by Saccharomyces cerevisiae exert remarkable influence on the taste and flavour of Chinese Baijiu. In order to study the regulation mechanism of amino acid metabolism genes on higher alcohol production, eight recombinant strains with amino acid metabolism gene deletion were constructed. The growth, fermentation performance, higher alcohol production, and expression level of genes in recombinant and original α5 strains were determined. Results displayed that the total higher alcohol concentration in α5ΔGDH1 strain decreased by 27.31% to 348.68 mg/L compared with that of α5. The total content of higher alcohols in α5ΔCAN1 and α5ΔGAT1 strains increased by 211.44% and 28.36% to 1493.96 and 615.73 mg/L, respectively, compared with that of α5. This study is the first to report that the CAN1 and GAT1 genes have great influence on the generation of higher alcohols. The results demonstrated that amino acid metabolism plays a substantial role in the metabolism of higher alcohols by S. cerevisiae. Interestingly, we also found that gene knockout downregulated the expression levels of the knocked out gene and other genes in the recombinant strain and thus affected the formation of higher alcohols by S. cerevisiae. This study provides worthy insights for comprehending the metabolic mechanism of higher alcohols in S. cerevisiae for Baijiu fermentation.

Assessment of genotoxic chemicals using chemogenomic profiling based on gene-knockout library in Saccharomyces cerevisiae

2021 ◽  
pp. 105278
Author(s):  
Miao Guan ◽  
Zheng Zhu ◽  
Ying Jiang ◽  
Mingming Tian ◽  
Lu Yan ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Gene Knockout ◽  
Genotoxic Chemicals

Saccharomyces cerevisiae glucose signalling regulator Mth1p regulates the organellar Na+/H+ exchanger Nhx1p

2010 ◽  
Vol 432 (2) ◽  
pp. 343-352 ◽  
Author(s):  
Keiji Mitsui ◽  
Masafumi Matsushita ◽  
Hiroshi Kanazawa
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Carbon Source ◽  
Sole Carbon Source ◽  
Glucose Sensor ◽  
Ph Regulation ◽  
Gene Knockout ◽  
Carbon Sources ◽  
Acidic Ph ◽  
In Cells

Organelle-localized NHEs (Na+/H+ exchangers) are found in cells from yeast to humans and contribute to organellar pH regulation by exporting H+ from the lumen to the cytosol coupled to an H+ gradient established by vacuolar H+-ATPase. The mechanisms underlying the regulation of organellar NHEs are largely unknown. In the present study, a yeast two-hybrid assay identified Mth1p as a new binding protein for Nhx1p, an organellar NHE in Saccharomyces cerevisiae. It was shown by an in vitro pull-down assay that Mth1p bound to the hydrophilic C-terminal half of Nhx1p, especially to the central portion of this region. Mth1p is known to bind to the cytoplasmic domain of the glucose sensor Snf3p/Rgt2p and also functions as a negative transcriptional regulator. Mth1p was expressed in cells grown in a medium containing galactose, but was lost (possibly degraded) when cells were grown in medium containing glucose as the sole carbon source. Deletion of the MTH1 gene increased cell growth compared with the wild-type when cells were grown in a medium containing galactose and with hygromycin or at an acidic pH. This resistance to hygromycin or acidic conditions was not observed for cells grown with glucose as the sole carbon source. Gene knockout of NHX1 increased the sensitivity to hygromycin and acidic pH. The increased resistance to hygromycin was reproduced by truncation of the Mth1p-binding region in Nhx1p. These results implicate Mth1p as a novel regulator of Nhx1p that responds to specific extracellular carbon sources.

scholarly journals Regulation of alpha-factor production in Saccharomyces cerevisiae: a-factor pheromone-induced expression of the MF alpha 1 and STE13 genes.

1989 ◽  
Vol 9 (10) ◽  
pp. 4507-4514 ◽  
Author(s):  
T Achstetter
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fusion Gene ◽  
Invertase Activity ◽  
Transcriptional Level ◽  
Alpha Cells ◽  
Mating Pheromone ◽  
Induced Expression ◽  
Alpha Factor ◽  
Suc2 Gene ◽  
Proteolytic Activities

Production of the mating pheromone alpha-factor was examined in Saccharomyces cerevisiae MAT alpha cells that had been exposed to the mating pheromone a-factor. A 2-h treatment with a-factor caused a significant increase in alpha-factor concentration in the medium as demonstrated by a halo assay. MF alpha 1 is one of the two genes coding for a precursor of alpha-factor. A Northern (RNA) analysis of total RNA from a-factor-treated MAT alpha cells revealed a rapid two- to threefold increase in MF alpha 1 transcript levels, reaching maximum within 60 min of exposure to the pheromone. Pheromone induction did not require ongoing protein synthesis. a-Factor-induced MF alpha 1 expression was quantitated by analysis of an MF alpha 1::SUC2 fusion gene whose product was assayed for invertase activity. Expression of the MF alpha 1::SUC2 gene in MAT alpha cells responded to the a-factor signal like the chromosomal version of MF alpha 1. Maturation of the alpha-factor precursor involves three proteolytic activities which are encoded by the KEX1, KEX2, and STE13 genes, respectively. Two of these genes, namely, KEX2 and STE13, were examined for pheromone-induced expression. Only the STE13 gene exhibited pheromone induction at the transcriptional level.

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