scholarly journals Isolation and Characterization of Live Yeast Cells from Ancient Clay Vessels

BIO-PROTOCOL ◽  
2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Tzemach Aouizerat ◽  
Aren Maeir ◽  
Yitzhak Paz ◽  
Yuval Gadot ◽  
Amir Szitenberg ◽  
...  
Keyword(s):  
Yeast Cells ◽  
Isolation And Characterization ◽  
Live Yeast

CHAPTER 11 ISOLATION AND CHARACTERIZATION OF LIVE YEAST CELLS FROM A MEAD VESSEL

2021 ◽  
pp. 131-141
Author(s):  
Tzemach Aouizerat ◽  
Itai Gutman ◽  
Yuval Gadot ◽  
Daniel Gelman ◽  
Amir Szitenberg ◽  
...  
Keyword(s):  
Yeast Cells ◽  
Isolation And Characterization ◽  
Live Yeast

ISOLATION AND CHARACTERIZATION OF LIVE YEAST CELLS FROM A MEAD VESSEL

2021 ◽  
pp. 131-141
Author(s):  
Tzemach Aouizerat ◽  
Itai Gutman ◽  
Yuval Gadot ◽  
Daniel Gelman ◽  
Amir Szitenberg ◽  
...  
Keyword(s):  
Yeast Cells ◽  
Isolation And Characterization ◽  
Live Yeast

scholarly journals Isolation and Characterization of Live Yeast Cells from Ancient Vessels as a Tool in Bio-Archaeology

mBio ◽  
2019 ◽  
Vol 10 (2) ◽  
Author(s):  
Tzemach Aouizerat ◽  
Itai Gutman ◽  
Yitzhak Paz ◽  
Aren M. Maeir ◽  
Yuval Gadot ◽  
...  
Keyword(s):  
Ancient Dna ◽  
Residue Analysis ◽  
Fermented Food ◽  
Yeast Cells ◽  
Fermented Beverages ◽  
Isolation And Characterization ◽  
Yeast Strains ◽  
Novel Approach ◽  
Clay Matrix

ABSTRACTAncient fermented food has been studied based on recipes, residue analysis, and ancient-DNA techniques and reconstructed using modern domesticated yeast. Here, we present a novel approach based on our hypothesis that enriched yeast populations in fermented beverages could have become the dominant species in storage vessels and their descendants could be isolated and studied today. We developed a pipeline of yeast isolation from clay vessels and screened for yeast cells in beverage-related and non-beverage-related ancient vessels and sediments from several archaeological sites. We found that yeast cells could be successfully isolated specifically from clay containers of fermented beverages. The findings that genotypically the isolated yeasts are similar to those found in traditional African beverages and phenotypically they grow similar to modern beer-producing yeast strongly suggest that they are descendants of the original fermenting yeast. These results demonstrate that modern microorganisms can serve as a new tool in bio-archaeology research.IMPORTANCESo far, most of the study of ancient organisms has been based mainly on the analysis of ancient DNA. Here we show that it is possible to isolate and study microorganisms—yeast in this case—from ancient pottery vessels used for fermentation. We demonstrate that it is highly likely that these cells are descendants of the original yeast strains that participated in the fermentation process and were absorbed into the clay matrix of the pottery vessels. Moreover, we characterized the isolated yeast strains, their genomes, and the beer they produced. These results open new and exciting avenues in the study of domesticated microorganisms and contribute significantly to the fields of bio- and experimental archaeology that aim to reconstruct ancient artifacts and products.

scholarly journals Isolation and Characterization of Cryptococcus neoformans Spores Reveal a Critical Role for Capsule Biosynthesis Genes in Spore Biogenesis

2009 ◽  
Vol 8 (4) ◽  
pp. 595-605 ◽  
Author(s):  
Michael R. Botts ◽  
Steven S. Giles ◽  
Marcellene A. Gates ◽  
Thomas R. Kozel ◽  
Christina M. Hull
Keyword(s):  
Cryptococcus Neoformans ◽  
Fungal Pathogens ◽  
Critical Role ◽  
Spore Formation ◽  
Yeast Cells ◽  
Isolation And Characterization ◽  
Specific Configuration ◽  
First Time

ABSTRACT Spores are essential particles for the survival of many organisms, both prokaryotic and eukaryotic. Among the eukaryotes, fungi have developed spores with superior resistance and dispersal properties. For the human fungal pathogens, however, relatively little is known about the role that spores play in dispersal and infection. Here we present the purification and characterization of spores from the environmental fungus Cryptococcus neoformans. For the first time, we purified spores to homogeneity and assessed their morphological, stress resistance, and surface properties. We found that spores are morphologically distinct from yeast cells and are covered with a thick spore coat. Spores are also more resistant to environmental stresses than yeast cells and display a spore-specific configuration of polysaccharides on their surfaces. Surprisingly, we found that the surface of the spore reacts with antibodies to the polysaccharide glucuronoxylomannan, the most abundant component of the polysaccharide capsule required for C. neoformans virulence. We explored the role of capsule polysaccharide in spore development by assessing spore formation in a series of acapsular strains and determined that capsule biosynthesis genes are required for proper sexual development and normal spore formation. Our findings suggest that C. neoformans spores may have an adapted cell surface that facilitates persistence in harsh environments and ultimately allows them to infect mammalian hosts.

scholarly journals Isolation and characterization of MOD5, a gene required for isopentenylation of cytoplasmic and mitochondrial tRNAs of Saccharomyces cerevisiae.

1987 ◽  
Vol 7 (1) ◽  
pp. 177-184 ◽  
Author(s):  
M E Dihanich ◽  
D Najarian ◽  
R Clark ◽  
E C Gillman ◽  
N C Martin ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Yeast Cells ◽  
Transferase Activity ◽  
Isopentenyl Transferase ◽  
Nuclear Mutation ◽  
Isolation And Characterization ◽  
Isopentenyl Adenosine ◽  
Cellular Compartments

The mod5-1 mutation is a nuclear mutation in Saccharomyces cerevisiae that reduces the biosynthesis of N6-(delta 2-isopentenyl)adenosine in both cytoplasmic and mitochondrial tRNAs to less than 1.5% of wild-type levels. The tRNA modification enzyme, delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase, cannot be detected in vitro with extracts from mod5-1 cells. A characterization of the MOD5 gene would help to determine how the same enzyme activity in different cellular compartments can be abolished by a single nuclear mutation. To that end we have cloned the MOD5 gene and shown that it restores delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase activity and N6-(delta 2-isopentenyl)adenosine to tRNA in both the mitochondria and the nucleus/cytoplasm compartments of mod5-1 yeast cells. That MOD5 sequences are expressed in Escherichia coli and can complement an N6-(delta 2-isopentenyl)-2-methylthioadenosine-deficient E. coli mutant leads us to conclude that MOD5 is the structural gene for delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase.

Isolation and characterization of a cDNA encoding a Δ8 sphingolipid desaturase from Aquilegia vulgaris

2002 ◽  
Vol 30 (6) ◽  
pp. 1073-1075 ◽  
Author(s):  
L. V. Michaelson ◽  
A. J. Longman ◽  
O. Sayanova ◽  
A. K. Stobart ◽  
J. A. Napier
Keyword(s):  
Functional Characterization ◽  
Yeast Cells ◽  
Sphingoid Bases ◽  
Yeast Saccharomyces Cerevisiae ◽  
Isolation And Characterization ◽  
Long Chain Base ◽  
In The Wild ◽  
Cis And Trans ◽  
Long Chain

We have isolated a cDNA encoding the Δ8 sphingolipid desaturase from the plant Aquilegia vulgaris L. via a PCR-based strategy using primers designed to target the conserved histidine box regions of microsomal desaturases. The function of the cDNA was confirmed by expression in the yeast, Saccharomyces cerevisiae. Analysis of the long-chain sphingoid bases as their dinitrophenyl derivatives by reverse-phase HPLC demonstrated the accumulation of cis- and trans-desaturated sphingoid bases which were not present in the wild-type yeast cells. The Δ8 desaturated products co-eluted with known Δ8-desaturated phytosphingenine and the molecular mass of these products was confirmed by liquid chromatography-MS. The Δ8 long-chain base desaturase was also able to desaturate dihydrosphingosine substrates. This is the first report of the functional characterization of an A. vulgaris gene product.

scholarly journals Isolation and Characterization of Senescent Cryptococcus neoformans and Implications for Phenotypic Switching and Pathogenesis in Chronic Cryptococcosis

2009 ◽  
Vol 8 (6) ◽  
pp. 858-866 ◽  
Author(s):  
Neena Jain ◽  
Emily Cook ◽  
Immaculata Xess ◽  
Fahmi Hasan ◽  
Dietrich Fries ◽  
...  
Keyword(s):  
Mathematical Modeling ◽  
Cryptococcus Neoformans ◽  
Chronic Infection ◽  
Yeast Cells ◽  
Phenotypic Switching ◽  
Isolation And Characterization ◽  
Immunocompromised Hosts ◽  
Infection Experiments

ABSTRACT Although several virulence factors and associated genes have been identified, the mechanisms that allow Cryptococcus neoformans to adapt during chronic infection and to persist in immunocompromised hosts remain poorly understood. Characterization of senescent cells of C. neoformans demonstrated that these cells exhibit a significantly enlarged cell body and capsule but still cross the blood-brain barrier. C. neoformans cells with advanced generational age are also more resistant to phagocytosis and killing by antifungals, which could promote their selection during chronic disease in humans. Senescent cells of RC-2, a C. neoformans strain that undergoes phenotypic switching, manifest switching rates up to 11-fold higher than those of younger cells. Infection experiments with labeled cells suggest that senescent yeast cells can potentially accumulate in vivo. Mathematical modeling incorporating different switching rates demonstrates how increased switching rates promote the emergence of hypervirulent mucoid variants during chronic infection. Our findings introduce the intriguing concept that senescence in eukaryotic pathogens could be a mechanism of microevolution that may promote pathoadaptation and facilitate evasion of an evolving immune response.

scholarly journals Isolation and characterization of live yeast cells from ancient vessels as a tool in bio-archeology

2019 ◽  
Author(s):  
Tzemach Aouizerat ◽  
Itai Gutman ◽  
Yitzhak Paz ◽  
Aren M. Maeir ◽  
Yuval Gadot ◽  
...  
Keyword(s):  
Ancient Dna ◽  
Residue Analysis ◽  
Genomic Analysis ◽  
Fermented Food ◽  
Yeast Cells ◽  
Fermented Beverages ◽  
Isolation And Characterization ◽  
Novel Approach ◽  
Experimental Archeology

AbstractAncient fermented food has been studied based on recipes, residue analysis and ancient-DNA techniques and reconstructed using modern domesticated yeast. Here, we present a novel approach. We hypothesize that enriched yeast populations in fermented beverages could have become the dominant species in storage vessels and the descendants of these yeast could be isolated and studied long after. To this end, using a pipeline of yeast isolation from clay vessels developed here, we screened for yeast cells in beverage-related and non-related ancient vessels and sediments, from several archeological sites. We found that yeast cells could be successfully isolated specifically from clay containers of fermented beverages. Genomic analysis revealed that these yeast are similar to those found in traditional African beverages. Phenotypically, they grow similar to modern-beer producing yeast. Both strongly suggesting that they are descendants of the original fermenting yeast. These findings provide modern microorganisms as a new tool in bio-archeology.ImportanceSo far, most of the study of ancient organisms was based mainly on the analysis of ancient DNA. Here we show that it is possible to isolate and study microorganisms, yeast in this case, from thousands of years old clay vessels, used for fermentation. We demonstrate that it is highly likely that these cells are descendants of the original yeast strains which participated in the fermentation process and were absorbed into the pottery vessels. Moreover, we characterize the isolated yeast their genome and the beer they produce. These results open new and exciting avenues in the study of domesticated microorganisms and contribute significantly to the fields of bio and experimental –archeology that aims to reconstruct ancient artifacts and products.

Isolation and characterization of MOD5, a gene required for isopentenylation of cytoplasmic and mitochondrial tRNAs of Saccharomyces cerevisiae

1987 ◽  
Vol 7 (1) ◽  
pp. 177-184
Author(s):  
M E Dihanich ◽  
D Najarian ◽  
R Clark ◽  
E C Gillman ◽  
N C Martin ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Yeast Cells ◽  
Transferase Activity ◽  
Isopentenyl Transferase ◽  
Nuclear Mutation ◽  
Isolation And Characterization ◽  
Isopentenyl Adenosine ◽  
Cellular Compartments

The mod5-1 mutation is a nuclear mutation in Saccharomyces cerevisiae that reduces the biosynthesis of N6-(delta 2-isopentenyl)adenosine in both cytoplasmic and mitochondrial tRNAs to less than 1.5% of wild-type levels. The tRNA modification enzyme, delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase, cannot be detected in vitro with extracts from mod5-1 cells. A characterization of the MOD5 gene would help to determine how the same enzyme activity in different cellular compartments can be abolished by a single nuclear mutation. To that end we have cloned the MOD5 gene and shown that it restores delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase activity and N6-(delta 2-isopentenyl)adenosine to tRNA in both the mitochondria and the nucleus/cytoplasm compartments of mod5-1 yeast cells. That MOD5 sequences are expressed in Escherichia coli and can complement an N6-(delta 2-isopentenyl)-2-methylthioadenosine-deficient E. coli mutant leads us to conclude that MOD5 is the structural gene for delta 2-isopentenyl pyrophosphate:tRNA isopentenyl transferase.

The isolation and characterization of 18 equine microsatellite loci, TKY272–TKY289

2000 ◽  
Vol 31 (2) ◽  
pp. 149-149 ◽  
Author(s):  
T Tozaki ◽  
H Kakoi ◽  
S Mashima ◽  
K Hirota ◽  
T Hasegawa ◽  
...  
Keyword(s):  
Microsatellite Loci ◽  
Isolation And Characterization
Sign in / Sign up

Export Citation Format

Share Document