Comet assay on tetraploid yeast cells
Tetraploid yeast cells ( Saccharomyces cerevisiae) were used in the comet assay with the intention of developing a new, fast and easy assay for detecting environmental genotoxic agents without using higher organisms. Two DNA-damaging chemicals, H 2O 2 and acrylamide, together with wastewater from th...
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| Veröffentlicht in: | Mutation research 2009-02, Vol.673 (1), p.53-58 |
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| creator | Rank, Jette Syberg, Kristian Jensen, Klara |
| description | Tetraploid yeast cells (
Saccharomyces cerevisiae) were used in the comet assay with the intention of developing a new, fast and easy assay for detecting environmental genotoxic agents without using higher organisms. Two DNA-damaging chemicals, H
2O
2 and acrylamide, together with wastewater from three municipal treatment plants were tested for their effect on the yeast-cell DNA. The main problem with using yeast in the comet assay is the necessity to degrade the cell wall. This was achieved by using Zymolase 100
T twice during the procedure, since Zymolase 20
T did not open the cell wall. Analytical problems that arose due to the small amount of DNA in the yeast nuclei in haploid and diploid cells, which contain 13
Mbp and 26
Mbp DNA per cell, respectively, were solved by using tetraploid yeast cells (52
Mbp) instead. DNA damage was shown after exposure to H
2O
2 and acrylamide. The lowest dose causing significant DNA damage was 20
μM for H
2O
2 and 200
mg/l for acrylamide. Tertiary-treated wastewater from the outlets of three municipal wastewater-treatment plants was tested, but did not cause DNA damage. Even though it is possible to produce comets with tetraploid yeast cells, the amount of DNA is likely too small for a proper comet assay. |
| doi_str_mv | 10.1016/j.mrgentox.2008.11.014 |
| format | Article |
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Saccharomyces cerevisiae) were used in the comet assay with the intention of developing a new, fast and easy assay for detecting environmental genotoxic agents without using higher organisms. Two DNA-damaging chemicals, H
2O
2 and acrylamide, together with wastewater from three municipal treatment plants were tested for their effect on the yeast-cell DNA. The main problem with using yeast in the comet assay is the necessity to degrade the cell wall. This was achieved by using Zymolase 100
T twice during the procedure, since Zymolase 20
T did not open the cell wall. Analytical problems that arose due to the small amount of DNA in the yeast nuclei in haploid and diploid cells, which contain 13
Mbp and 26
Mbp DNA per cell, respectively, were solved by using tetraploid yeast cells (52
Mbp) instead. DNA damage was shown after exposure to H
2O
2 and acrylamide. The lowest dose causing significant DNA damage was 20
μM for H
2O
2 and 200
mg/l for acrylamide. Tertiary-treated wastewater from the outlets of three municipal wastewater-treatment plants was tested, but did not cause DNA damage. Even though it is possible to produce comets with tetraploid yeast cells, the amount of DNA is likely too small for a proper comet assay.</description><identifier>ISSN: 1383-5718</identifier><identifier>ISSN: 0027-5107</identifier><identifier>EISSN: 1879-3592</identifier><identifier>DOI: 10.1016/j.mrgentox.2008.11.014</identifier><identifier>PMID: 19146983</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Acrylamide ; Acrylamide - pharmacology ; Biological and medical sciences ; Cell Wall - drug effects ; Comet Assay - methods ; DNA damage ; DNA Damage - drug effects ; Fundamental and applied biological sciences. Psychology ; Genetics of eukaryotes. Biological and molecular evolution ; Genotoxicity ; H 2O 2 ; Hydrogen Peroxide - pharmacology ; Medical sciences ; Polyploidy ; Saccharomyces cerevisiae ; Toxicology ; Waste Products ; Wastewater ; Yeasts - drug effects ; Yeasts - metabolism ; Zymolase</subject><ispartof>Mutation research, 2009-02, Vol.673 (1), p.53-58</ispartof><rights>2008 Elsevier B.V.</rights><rights>2009 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c427t-8b64237f09ad483a00a83c121eac75f02510dd149aa26767416bee84b85f688a3</citedby><cites>FETCH-LOGICAL-c427t-8b64237f09ad483a00a83c121eac75f02510dd149aa26767416bee84b85f688a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.mrgentox.2008.11.014$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=21146212$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19146983$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rank, Jette</creatorcontrib><creatorcontrib>Syberg, Kristian</creatorcontrib><creatorcontrib>Jensen, Klara</creatorcontrib><title>Comet assay on tetraploid yeast cells</title><title>Mutation research</title><addtitle>Mutat Res</addtitle><description>Tetraploid yeast cells (
Saccharomyces cerevisiae) were used in the comet assay with the intention of developing a new, fast and easy assay for detecting environmental genotoxic agents without using higher organisms. Two DNA-damaging chemicals, H
2O
2 and acrylamide, together with wastewater from three municipal treatment plants were tested for their effect on the yeast-cell DNA. The main problem with using yeast in the comet assay is the necessity to degrade the cell wall. This was achieved by using Zymolase 100
T twice during the procedure, since Zymolase 20
T did not open the cell wall. Analytical problems that arose due to the small amount of DNA in the yeast nuclei in haploid and diploid cells, which contain 13
Mbp and 26
Mbp DNA per cell, respectively, were solved by using tetraploid yeast cells (52
Mbp) instead. DNA damage was shown after exposure to H
2O
2 and acrylamide. The lowest dose causing significant DNA damage was 20
μM for H
2O
2 and 200
mg/l for acrylamide. Tertiary-treated wastewater from the outlets of three municipal wastewater-treatment plants was tested, but did not cause DNA damage. Even though it is possible to produce comets with tetraploid yeast cells, the amount of DNA is likely too small for a proper comet assay.</description><subject>Acrylamide</subject><subject>Acrylamide - pharmacology</subject><subject>Biological and medical sciences</subject><subject>Cell Wall - drug effects</subject><subject>Comet Assay - methods</subject><subject>DNA damage</subject><subject>DNA Damage - drug effects</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetics of eukaryotes. Biological and molecular evolution</subject><subject>Genotoxicity</subject><subject>H 2O 2</subject><subject>Hydrogen Peroxide - pharmacology</subject><subject>Medical sciences</subject><subject>Polyploidy</subject><subject>Saccharomyces cerevisiae</subject><subject>Toxicology</subject><subject>Waste Products</subject><subject>Wastewater</subject><subject>Yeasts - drug effects</subject><subject>Yeasts - metabolism</subject><subject>Zymolase</subject><issn>1383-5718</issn><issn>0027-5107</issn><issn>1879-3592</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkMtOwzAQRS0EoqXwC1U2ZZfgsZ3E2YEqXlIlNrC2Js4Epcqj2Cmif4-rFliymlmcO3N1GJsDT4BDdrNOOvdO_Th8JYJznQAkHNQJm4LOi1imhTgNu9QyTnPQE3bh_ZpzwSXX52wCBais0HLKFsuhozFC73EXDX000uhw0w5NFe0I_RhZalt_yc5qbD1dHeeMvT3cvy6f4tXL4_PybhVbJfIx1mWmhMxrXmCltETOUUsLAghtntZcpMCrClSBKLI8yxVkJZFWpU7rTGuUM3Z9uLtxw8eW_Gi6xu8bYE_D1hvBQy4t0gBmB9C6wXtHtdm4pkO3M8DNXpBZmx9BZi_IAJggKATnxw_bsqPqL3Y0EoDFEUBvsa0d9rbxv5yAwAkQgbs9cBR8fDbkjLcN9ZaqxpEdTTU0_3X5Bh7bhbU</recordid><startdate>20090219</startdate><enddate>20090219</enddate><creator>Rank, Jette</creator><creator>Syberg, Kristian</creator><creator>Jensen, Klara</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>20090219</creationdate><title>Comet assay on tetraploid yeast cells</title><author>Rank, Jette ; Syberg, Kristian ; Jensen, Klara</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c427t-8b64237f09ad483a00a83c121eac75f02510dd149aa26767416bee84b85f688a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Acrylamide</topic><topic>Acrylamide - pharmacology</topic><topic>Biological and medical sciences</topic><topic>Cell Wall - drug effects</topic><topic>Comet Assay - methods</topic><topic>DNA damage</topic><topic>DNA Damage - drug effects</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetics of eukaryotes. Biological and molecular evolution</topic><topic>Genotoxicity</topic><topic>H 2O 2</topic><topic>Hydrogen Peroxide - pharmacology</topic><topic>Medical sciences</topic><topic>Polyploidy</topic><topic>Saccharomyces cerevisiae</topic><topic>Toxicology</topic><topic>Waste Products</topic><topic>Wastewater</topic><topic>Yeasts - drug effects</topic><topic>Yeasts - metabolism</topic><topic>Zymolase</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rank, Jette</creatorcontrib><creatorcontrib>Syberg, Kristian</creatorcontrib><creatorcontrib>Jensen, Klara</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Mutation research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rank, Jette</au><au>Syberg, Kristian</au><au>Jensen, Klara</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comet assay on tetraploid yeast cells</atitle><jtitle>Mutation research</jtitle><addtitle>Mutat Res</addtitle><date>2009-02-19</date><risdate>2009</risdate><volume>673</volume><issue>1</issue><spage>53</spage><epage>58</epage><pages>53-58</pages><issn>1383-5718</issn><issn>0027-5107</issn><eissn>1879-3592</eissn><abstract>Tetraploid yeast cells (
Saccharomyces cerevisiae) were used in the comet assay with the intention of developing a new, fast and easy assay for detecting environmental genotoxic agents without using higher organisms. Two DNA-damaging chemicals, H
2O
2 and acrylamide, together with wastewater from three municipal treatment plants were tested for their effect on the yeast-cell DNA. The main problem with using yeast in the comet assay is the necessity to degrade the cell wall. This was achieved by using Zymolase 100
T twice during the procedure, since Zymolase 20
T did not open the cell wall. Analytical problems that arose due to the small amount of DNA in the yeast nuclei in haploid and diploid cells, which contain 13
Mbp and 26
Mbp DNA per cell, respectively, were solved by using tetraploid yeast cells (52
Mbp) instead. DNA damage was shown after exposure to H
2O
2 and acrylamide. The lowest dose causing significant DNA damage was 20
μM for H
2O
2 and 200
mg/l for acrylamide. Tertiary-treated wastewater from the outlets of three municipal wastewater-treatment plants was tested, but did not cause DNA damage. Even though it is possible to produce comets with tetraploid yeast cells, the amount of DNA is likely too small for a proper comet assay.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>19146983</pmid><doi>10.1016/j.mrgentox.2008.11.014</doi><tpages>6</tpages></addata></record> |
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| ispartof | Mutation research, 2009-02, Vol.673 (1), p.53-58 |
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| subjects | Acrylamide Acrylamide - pharmacology Biological and medical sciences Cell Wall - drug effects Comet Assay - methods DNA damage DNA Damage - drug effects Fundamental and applied biological sciences. Psychology Genetics of eukaryotes. Biological and molecular evolution Genotoxicity H 2O 2 Hydrogen Peroxide - pharmacology Medical sciences Polyploidy Saccharomyces cerevisiae Toxicology Waste Products Wastewater Yeasts - drug effects Yeasts - metabolism Zymolase |
| title | Comet assay on tetraploid yeast cells |
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