Molecular cloning, characterization of copper/zinc superoxide dismutase and expression analysis of stress-responsive genes from Eisenia fetida against dietary zinc oxide
The full length cDNA of copper/zinc superoxide dismutase (Cu/Zn-SOD) from Eisenia fetida (E. fetida) was cloned (GenBank accession no. JN579648). Sequence characterization revealed that the cDNA contained characteristic Cu/Zn-SOD family signatures (45GFHVHEFGDNT55 and 138GNAGGRLACGVI149), cysteines...
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| Veröffentlicht in: | Comparative biochemistry and physiology. Toxicology & pharmacology 2012-03, Vol.155 (2), p.416-422 |
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| creator | Xiong, Wenguang Bai, Ling Muhammad, Rizwan-Ul-Haq Zou, Mengjia Sun, Yongxue |
| description | The full length cDNA of copper/zinc superoxide dismutase (Cu/Zn-SOD) from Eisenia fetida (E. fetida) was cloned (GenBank accession no. JN579648). Sequence characterization revealed that the cDNA contained characteristic Cu/Zn-SOD family signatures (45GFHVHEFGDNT55 and 138GNAGGRLACGVI149), cysteines (Cys-58 and-146) predicted to form one disulphide bond, Cu-binding (His-47, -49, -64 and -120) and Zn-binding (His-64, -72, -81 and Asp-84). They were essential for the structure and function of Cu/Zn-SOD. Differential expression of stress-responsive genes like Cu/Zn-SOD, catalase (CAT), heat shock protein 70 (Hsp70) and metallothionein (MT) was applied as potential biomarkers to assess their efficacy for the ecotoxicological effects of dietary zinc oxide (ZnO) on E. fetida. The results showed that the expression of Cu/Zn-SOD and MT increased to reach the highest levels of 6.22 and 7.68 fold in a dose-dependent manner at day 10 respectively. The highest expression of 3.03 fold of CAT was registered at day 10. The transient expression of Hsp70 without consistent time- or/and dose-dependent was observed. It implied that the transcriptional patterns of Cu/Zn-SOD, CAT and MT could serve as early warning signals in ecotoxicological assessment of dietary ZnO on earthworms while the expression of Hsp70 was not well done, which is helpful to monitoring and regulation of ZnO in veterinary application. |
| doi_str_mv | 10.1016/j.cbpc.2011.11.004 |
| format | Article |
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JN579648). Sequence characterization revealed that the cDNA contained characteristic Cu/Zn-SOD family signatures (45GFHVHEFGDNT55 and 138GNAGGRLACGVI149), cysteines (Cys-58 and-146) predicted to form one disulphide bond, Cu-binding (His-47, -49, -64 and -120) and Zn-binding (His-64, -72, -81 and Asp-84). They were essential for the structure and function of Cu/Zn-SOD. Differential expression of stress-responsive genes like Cu/Zn-SOD, catalase (CAT), heat shock protein 70 (Hsp70) and metallothionein (MT) was applied as potential biomarkers to assess their efficacy for the ecotoxicological effects of dietary zinc oxide (ZnO) on E. fetida. The results showed that the expression of Cu/Zn-SOD and MT increased to reach the highest levels of 6.22 and 7.68 fold in a dose-dependent manner at day 10 respectively. The highest expression of 3.03 fold of CAT was registered at day 10. The transient expression of Hsp70 without consistent time- or/and dose-dependent was observed. It implied that the transcriptional patterns of Cu/Zn-SOD, CAT and MT could serve as early warning signals in ecotoxicological assessment of dietary ZnO on earthworms while the expression of Hsp70 was not well done, which is helpful to monitoring and regulation of ZnO in veterinary application.</description><identifier>ISSN: 1532-0456</identifier><identifier>EISSN: 1878-1659</identifier><identifier>DOI: 10.1016/j.cbpc.2011.11.004</identifier><identifier>PMID: 22137962</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acid Sequence ; Animals ; Base Sequence ; Biomarker ; biomarkers ; Catalase ; Catalase - genetics ; cDNA cloning ; Chromium ; Cloning, Molecular ; complementary DNA ; Copper ; Cysteine ; Diet ; Dietary ZnO ; disulfide bonds ; DNA, Complementary - chemistry ; DNA, Complementary - genetics ; dose response ; earthworms ; Ecotoxicological ; Eisenia fetida ; Gene Expression - drug effects ; gene expression regulation ; genes ; heat-shock protein 70 ; HSP70 Heat-Shock Proteins - genetics ; Hsp70 protein ; Metallothionein ; Metallothionein - genetics ; molecular cloning ; Molecular Sequence Data ; monitoring ; mRNA expression ; nutrition assessment ; Oligochaeta - enzymology ; Oligochaeta - genetics ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Stress, Physiological - genetics ; Structure-function relationships ; Superoxide dismutase ; Superoxide Dismutase - classification ; Superoxide Dismutase - genetics ; Toxicity Tests - methods ; Transcription ; transcription (genetics) ; Zinc ; zinc oxide ; Zinc Oxide - administration & dosage ; Zinc Oxide - toxicity</subject><ispartof>Comparative biochemistry and physiology. Toxicology & pharmacology, 2012-03, Vol.155 (2), p.416-422</ispartof><rights>2011 Elsevier Inc.</rights><rights>Copyright © 2011 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c411t-59cc9f2b9ac290e5b6adab53a3046dbdcfd8a5ad690b593805e9b96231e68b3</citedby><cites>FETCH-LOGICAL-c411t-59cc9f2b9ac290e5b6adab53a3046dbdcfd8a5ad690b593805e9b96231e68b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.cbpc.2011.11.004$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27922,27923,45993</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22137962$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Xiong, Wenguang</creatorcontrib><creatorcontrib>Bai, Ling</creatorcontrib><creatorcontrib>Muhammad, Rizwan-Ul-Haq</creatorcontrib><creatorcontrib>Zou, Mengjia</creatorcontrib><creatorcontrib>Sun, Yongxue</creatorcontrib><title>Molecular cloning, characterization of copper/zinc superoxide dismutase and expression analysis of stress-responsive genes from Eisenia fetida against dietary zinc oxide</title><title>Comparative biochemistry and physiology. Toxicology & pharmacology</title><addtitle>Comp Biochem Physiol C Toxicol Pharmacol</addtitle><description>The full length cDNA of copper/zinc superoxide dismutase (Cu/Zn-SOD) from Eisenia fetida (E. fetida) was cloned (GenBank accession no. JN579648). Sequence characterization revealed that the cDNA contained characteristic Cu/Zn-SOD family signatures (45GFHVHEFGDNT55 and 138GNAGGRLACGVI149), cysteines (Cys-58 and-146) predicted to form one disulphide bond, Cu-binding (His-47, -49, -64 and -120) and Zn-binding (His-64, -72, -81 and Asp-84). They were essential for the structure and function of Cu/Zn-SOD. Differential expression of stress-responsive genes like Cu/Zn-SOD, catalase (CAT), heat shock protein 70 (Hsp70) and metallothionein (MT) was applied as potential biomarkers to assess their efficacy for the ecotoxicological effects of dietary zinc oxide (ZnO) on E. fetida. The results showed that the expression of Cu/Zn-SOD and MT increased to reach the highest levels of 6.22 and 7.68 fold in a dose-dependent manner at day 10 respectively. The highest expression of 3.03 fold of CAT was registered at day 10. The transient expression of Hsp70 without consistent time- or/and dose-dependent was observed. It implied that the transcriptional patterns of Cu/Zn-SOD, CAT and MT could serve as early warning signals in ecotoxicological assessment of dietary ZnO on earthworms while the expression of Hsp70 was not well done, which is helpful to monitoring and regulation of ZnO in veterinary application.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Biomarker</subject><subject>biomarkers</subject><subject>Catalase</subject><subject>Catalase - genetics</subject><subject>cDNA cloning</subject><subject>Chromium</subject><subject>Cloning, Molecular</subject><subject>complementary DNA</subject><subject>Copper</subject><subject>Cysteine</subject><subject>Diet</subject><subject>Dietary ZnO</subject><subject>disulfide bonds</subject><subject>DNA, Complementary - chemistry</subject><subject>DNA, Complementary - genetics</subject><subject>dose response</subject><subject>earthworms</subject><subject>Ecotoxicological</subject><subject>Eisenia fetida</subject><subject>Gene Expression - drug effects</subject><subject>gene expression regulation</subject><subject>genes</subject><subject>heat-shock protein 70</subject><subject>HSP70 Heat-Shock Proteins - genetics</subject><subject>Hsp70 protein</subject><subject>Metallothionein</subject><subject>Metallothionein - genetics</subject><subject>molecular cloning</subject><subject>Molecular Sequence Data</subject><subject>monitoring</subject><subject>mRNA expression</subject><subject>nutrition assessment</subject><subject>Oligochaeta - enzymology</subject><subject>Oligochaeta - genetics</subject><subject>Phylogeny</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Sequence Analysis, DNA</subject><subject>Sequence Homology, Amino Acid</subject><subject>Stress, Physiological - genetics</subject><subject>Structure-function relationships</subject><subject>Superoxide dismutase</subject><subject>Superoxide Dismutase - classification</subject><subject>Superoxide Dismutase - genetics</subject><subject>Toxicity Tests - methods</subject><subject>Transcription</subject><subject>transcription (genetics)</subject><subject>Zinc</subject><subject>zinc oxide</subject><subject>Zinc Oxide - administration & dosage</subject><subject>Zinc Oxide - toxicity</subject><issn>1532-0456</issn><issn>1878-1659</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc1u1TAQhSMEoqXwAizAOzbk1k5iN5bYVFX5kYpYFNbWxJ5cfJVrB49TtX0j3hKnt7BEsuyRdc6Z0XxV9VrwjeBCne42dpjtpuFCbMrhvHtSHYv-rK-FkvppqWXb1LyT6qh6QbTjnMtOqOfVUdOI9kyr5rj6_TVOaJcJErNTDD5s3zP7ExLYjMnfQ_YxsDgyG-cZ0-m9D5bRUsp46x0y52m_ZCBkEBzD2zkh0WqBANMdeVq9lNffulxzDORvkG0xILExxT279ITBAxsxewcMtuAD5RKMGdIde2j40Otl9WyEifDV43tSXX-8_H7xub769unLxflVbTshci21tXpsBg220RzloMDBIFtoeafc4OzoepDglOaD1G3PJeqhrKIVqPqhPaneHVLnFH8tSNnsPVmcJggYFzK64X0JatuibA5KmyJRwtHMye_LzEZws_IxO7PyMSsfU07hU0xvHuOXYY_un-UvkCJ4exCMEA1skyfz47okqAKvk7xfIz4cFFiWcOMxGbIeg0XnE9psXPT_m-AP--OwOw</recordid><startdate>20120301</startdate><enddate>20120301</enddate><creator>Xiong, Wenguang</creator><creator>Bai, Ling</creator><creator>Muhammad, Rizwan-Ul-Haq</creator><creator>Zou, Mengjia</creator><creator>Sun, Yongxue</creator><general>Elsevier Inc</general><scope>FBQ</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>P64</scope><scope>RC3</scope></search><sort><creationdate>20120301</creationdate><title>Molecular cloning, characterization of copper/zinc superoxide dismutase and expression analysis of stress-responsive genes from Eisenia fetida against dietary zinc oxide</title><author>Xiong, Wenguang ; Bai, Ling ; Muhammad, Rizwan-Ul-Haq ; Zou, Mengjia ; Sun, Yongxue</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c411t-59cc9f2b9ac290e5b6adab53a3046dbdcfd8a5ad690b593805e9b96231e68b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Biomarker</topic><topic>biomarkers</topic><topic>Catalase</topic><topic>Catalase - genetics</topic><topic>cDNA cloning</topic><topic>Chromium</topic><topic>Cloning, Molecular</topic><topic>complementary DNA</topic><topic>Copper</topic><topic>Cysteine</topic><topic>Diet</topic><topic>Dietary ZnO</topic><topic>disulfide bonds</topic><topic>DNA, Complementary - chemistry</topic><topic>DNA, Complementary - genetics</topic><topic>dose response</topic><topic>earthworms</topic><topic>Ecotoxicological</topic><topic>Eisenia fetida</topic><topic>Gene Expression - drug effects</topic><topic>gene expression regulation</topic><topic>genes</topic><topic>heat-shock protein 70</topic><topic>HSP70 Heat-Shock Proteins - genetics</topic><topic>Hsp70 protein</topic><topic>Metallothionein</topic><topic>Metallothionein - genetics</topic><topic>molecular cloning</topic><topic>Molecular Sequence Data</topic><topic>monitoring</topic><topic>mRNA expression</topic><topic>nutrition assessment</topic><topic>Oligochaeta - enzymology</topic><topic>Oligochaeta - genetics</topic><topic>Phylogeny</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Sequence Analysis, DNA</topic><topic>Sequence Homology, Amino Acid</topic><topic>Stress, Physiological - genetics</topic><topic>Structure-function relationships</topic><topic>Superoxide dismutase</topic><topic>Superoxide Dismutase - classification</topic><topic>Superoxide Dismutase - genetics</topic><topic>Toxicity Tests - methods</topic><topic>Transcription</topic><topic>transcription (genetics)</topic><topic>Zinc</topic><topic>zinc oxide</topic><topic>Zinc Oxide - administration & dosage</topic><topic>Zinc Oxide - toxicity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Xiong, Wenguang</creatorcontrib><creatorcontrib>Bai, Ling</creatorcontrib><creatorcontrib>Muhammad, Rizwan-Ul-Haq</creatorcontrib><creatorcontrib>Zou, Mengjia</creatorcontrib><creatorcontrib>Sun, Yongxue</creatorcontrib><collection>AGRIS</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>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Comparative biochemistry and physiology. Toxicology & pharmacology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xiong, Wenguang</au><au>Bai, Ling</au><au>Muhammad, Rizwan-Ul-Haq</au><au>Zou, Mengjia</au><au>Sun, Yongxue</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular cloning, characterization of copper/zinc superoxide dismutase and expression analysis of stress-responsive genes from Eisenia fetida against dietary zinc oxide</atitle><jtitle>Comparative biochemistry and physiology. Toxicology & pharmacology</jtitle><addtitle>Comp Biochem Physiol C Toxicol Pharmacol</addtitle><date>2012-03-01</date><risdate>2012</risdate><volume>155</volume><issue>2</issue><spage>416</spage><epage>422</epage><pages>416-422</pages><issn>1532-0456</issn><eissn>1878-1659</eissn><abstract>The full length cDNA of copper/zinc superoxide dismutase (Cu/Zn-SOD) from Eisenia fetida (E. fetida) was cloned (GenBank accession no. JN579648). Sequence characterization revealed that the cDNA contained characteristic Cu/Zn-SOD family signatures (45GFHVHEFGDNT55 and 138GNAGGRLACGVI149), cysteines (Cys-58 and-146) predicted to form one disulphide bond, Cu-binding (His-47, -49, -64 and -120) and Zn-binding (His-64, -72, -81 and Asp-84). They were essential for the structure and function of Cu/Zn-SOD. Differential expression of stress-responsive genes like Cu/Zn-SOD, catalase (CAT), heat shock protein 70 (Hsp70) and metallothionein (MT) was applied as potential biomarkers to assess their efficacy for the ecotoxicological effects of dietary zinc oxide (ZnO) on E. fetida. The results showed that the expression of Cu/Zn-SOD and MT increased to reach the highest levels of 6.22 and 7.68 fold in a dose-dependent manner at day 10 respectively. The highest expression of 3.03 fold of CAT was registered at day 10. The transient expression of Hsp70 without consistent time- or/and dose-dependent was observed. It implied that the transcriptional patterns of Cu/Zn-SOD, CAT and MT could serve as early warning signals in ecotoxicological assessment of dietary ZnO on earthworms while the expression of Hsp70 was not well done, which is helpful to monitoring and regulation of ZnO in veterinary application.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>22137962</pmid><doi>10.1016/j.cbpc.2011.11.004</doi><tpages>7</tpages></addata></record> |
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| ispartof | Comparative biochemistry and physiology. Toxicology & pharmacology, 2012-03, Vol.155 (2), p.416-422 |
| issn | 1532-0456 1878-1659 |
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| subjects | Amino Acid Sequence Animals Base Sequence Biomarker biomarkers Catalase Catalase - genetics cDNA cloning Chromium Cloning, Molecular complementary DNA Copper Cysteine Diet Dietary ZnO disulfide bonds DNA, Complementary - chemistry DNA, Complementary - genetics dose response earthworms Ecotoxicological Eisenia fetida Gene Expression - drug effects gene expression regulation genes heat-shock protein 70 HSP70 Heat-Shock Proteins - genetics Hsp70 protein Metallothionein Metallothionein - genetics molecular cloning Molecular Sequence Data monitoring mRNA expression nutrition assessment Oligochaeta - enzymology Oligochaeta - genetics Phylogeny Reverse Transcriptase Polymerase Chain Reaction Sequence Analysis, DNA Sequence Homology, Amino Acid Stress, Physiological - genetics Structure-function relationships Superoxide dismutase Superoxide Dismutase - classification Superoxide Dismutase - genetics Toxicity Tests - methods Transcription transcription (genetics) Zinc zinc oxide Zinc Oxide - administration & dosage Zinc Oxide - toxicity |
| title | Molecular cloning, characterization of copper/zinc superoxide dismutase and expression analysis of stress-responsive genes from Eisenia fetida against dietary zinc oxide |
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