Resistance to cadmium and zinc in Acidiphilium symbioticum KM2 is plasmid mediated
The acidophilic heterotroph, Acidiphilium symbioticum KM2, is highly resistant to several metals and harbors three plasmids of 3.8, 7.1, and 56 kb in size. The bacterium becomes extremely sensitive to metals when it is cured of its plasmids. A mini-plasmid library was constructed by ligating the pla...
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| Veröffentlicht in: | Current microbiology 2002-09, Vol.45 (3), p.180-186 |
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| creator | MAHAPATRA, Nitish R GHOSH, Sajalendu DEB, Chirajyoti BANERJEE, Pataki C |
| description | The acidophilic heterotroph, Acidiphilium symbioticum KM2, is highly resistant to several metals and harbors three plasmids of 3.8, 7.1, and 56 kb in size. The bacterium becomes extremely sensitive to metals when it is cured of its plasmids. A mini-plasmid library was constructed by ligating the plasmid DNA fragments generated by MboI partial digestion into the BamHI site of pBluescriptII KS+. The Lac(-)Amp(r) transformants of Escherichia coli DH5alpha, isolated after transformation with the library, were counter-selected on Cu(2+), Cd(2+), Ni(2+), and Zn(2+)-containing plates. Only Cd(2+)- and Zn(2+)-resistant colonies were developed, and, after screening, four types of recombinant plasmids designated as pNM201 (7.2 kb), pNM206 (3.4 kb), pNM208 (4.5 kb), and pNM215 (4.9 kb) were obtained. The DNA insert in pNM206 hybridized strongly with the 3.8-kb plasmid and weakly with the 7.1-kb plasmid of Acidiphilium symbioticum KM2. The DNA insert in pNM215 hybridized only with the 7.1-kb plasmid. These results strongly suggested that resistance to cadmium and zinc in A. symbioticum KM2 is mediated by these plasmids. The smallest insert of 422 bp in pNM206 conferring metal resistance in E. coli has no sequence similarity with the reported metal-resistant genes. All the putative ORFs are significantly rich (up to 37%) in basic amino acids, mainly arginine. |
| doi_str_mv | 10.1007/s00284-001-0113-6 |
| format | Article |
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The bacterium becomes extremely sensitive to metals when it is cured of its plasmids. A mini-plasmid library was constructed by ligating the plasmid DNA fragments generated by MboI partial digestion into the BamHI site of pBluescriptII KS+. The Lac(-)Amp(r) transformants of Escherichia coli DH5alpha, isolated after transformation with the library, were counter-selected on Cu(2+), Cd(2+), Ni(2+), and Zn(2+)-containing plates. Only Cd(2+)- and Zn(2+)-resistant colonies were developed, and, after screening, four types of recombinant plasmids designated as pNM201 (7.2 kb), pNM206 (3.4 kb), pNM208 (4.5 kb), and pNM215 (4.9 kb) were obtained. The DNA insert in pNM206 hybridized strongly with the 3.8-kb plasmid and weakly with the 7.1-kb plasmid of Acidiphilium symbioticum KM2. The DNA insert in pNM215 hybridized only with the 7.1-kb plasmid. These results strongly suggested that resistance to cadmium and zinc in A. symbioticum KM2 is mediated by these plasmids. The smallest insert of 422 bp in pNM206 conferring metal resistance in E. coli has no sequence similarity with the reported metal-resistant genes. All the putative ORFs are significantly rich (up to 37%) in basic amino acids, mainly arginine.</description><identifier>ISSN: 0343-8651</identifier><identifier>EISSN: 1432-0991</identifier><identifier>DOI: 10.1007/s00284-001-0113-6</identifier><identifier>PMID: 12177739</identifier><identifier>CODEN: CUMIDD</identifier><language>eng</language><publisher>New York, NY: Springer</publisher><subject>Acetobacteraceae - drug effects ; Acetobacteraceae - genetics ; Acidiphilium ; Action of physical and chemical agents on bacteria ; Amino Acid Sequence ; Amino acids ; Bacteriology ; Base Sequence ; Biological and medical sciences ; Cadmium ; Cadmium - pharmacology ; Cloning, Molecular ; Deoxyribonucleic acid ; DNA ; DNA, Bacterial - genetics ; Drug Resistance, Bacterial - genetics ; E coli ; Escherichia coli - genetics ; Fundamental and applied biological sciences. Psychology ; Genetics ; Metals ; Microbiology ; Mining ; Molecular Sequence Data ; Plasmids ; Plasmids - genetics ; Transformation, Genetic ; Zinc ; Zinc - pharmacology</subject><ispartof>Current microbiology, 2002-09, Vol.45 (3), p.180-186</ispartof><rights>Springer-Verlag New York Inc. 2002</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c419t-25513da528a83113e505bfb7ef7dd0d4dde95e4ff61296fe6273c366a4bdccd03</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14655319$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12177739$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>MAHAPATRA, Nitish R</creatorcontrib><creatorcontrib>GHOSH, Sajalendu</creatorcontrib><creatorcontrib>DEB, Chirajyoti</creatorcontrib><creatorcontrib>BANERJEE, Pataki C</creatorcontrib><title>Resistance to cadmium and zinc in Acidiphilium symbioticum KM2 is plasmid mediated</title><title>Current microbiology</title><addtitle>Curr Microbiol</addtitle><description>The acidophilic heterotroph, Acidiphilium symbioticum KM2, is highly resistant to several metals and harbors three plasmids of 3.8, 7.1, and 56 kb in size. The bacterium becomes extremely sensitive to metals when it is cured of its plasmids. A mini-plasmid library was constructed by ligating the plasmid DNA fragments generated by MboI partial digestion into the BamHI site of pBluescriptII KS+. The Lac(-)Amp(r) transformants of Escherichia coli DH5alpha, isolated after transformation with the library, were counter-selected on Cu(2+), Cd(2+), Ni(2+), and Zn(2+)-containing plates. Only Cd(2+)- and Zn(2+)-resistant colonies were developed, and, after screening, four types of recombinant plasmids designated as pNM201 (7.2 kb), pNM206 (3.4 kb), pNM208 (4.5 kb), and pNM215 (4.9 kb) were obtained. The DNA insert in pNM206 hybridized strongly with the 3.8-kb plasmid and weakly with the 7.1-kb plasmid of Acidiphilium symbioticum KM2. The DNA insert in pNM215 hybridized only with the 7.1-kb plasmid. These results strongly suggested that resistance to cadmium and zinc in A. symbioticum KM2 is mediated by these plasmids. The smallest insert of 422 bp in pNM206 conferring metal resistance in E. coli has no sequence similarity with the reported metal-resistant genes. All the putative ORFs are significantly rich (up to 37%) in basic amino acids, mainly arginine.</description><subject>Acetobacteraceae - drug effects</subject><subject>Acetobacteraceae - genetics</subject><subject>Acidiphilium</subject><subject>Action of physical and chemical agents on bacteria</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Bacteriology</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cadmium</subject><subject>Cadmium - pharmacology</subject><subject>Cloning, Molecular</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Bacterial - genetics</subject><subject>Drug Resistance, Bacterial - genetics</subject><subject>E coli</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetics</subject><subject>Metals</subject><subject>Microbiology</subject><subject>Mining</subject><subject>Molecular Sequence Data</subject><subject>Plasmids</subject><subject>Plasmids - genetics</subject><subject>Transformation, Genetic</subject><subject>Zinc</subject><subject>Zinc - pharmacology</subject><issn>0343-8651</issn><issn>1432-0991</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>BENPR</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqFkc1qGzEUhUVIiZ00D5BNEISGbibV1e9oGUzSlroUTLMWGklDZObHGc0s3KePBhsCWaQrCfTdw9X5ELoCcgeEqG-JEFryghAoCAAr5AlaAme0IFrDKVoSxllRSgELdJ7SNnNUEzhDC6CglGJ6iTabkGIabecCHnvsrG_j1GLbefwvdg7HDt-76OPuOTbzQ9q3VezH6PL912-KY8K7xqY2etwGH-0Y_Gf0qbZNCpfH8wI9PT78Xf0o1n--_1zdrwvHQY8FFQKYt4KWtmR5-yCIqOpKhVp5Tzz3PmgReF3LvLWsg6SKOSal5ZV3zhN2gW4Pubuhf5lCGk0bkwtNY7vQT8komuuRiv4XhFJpzUvI4NePQc5LSnO5c-bNO3TbT0OX_5spBlopIDMFB8oNfUpDqM1uiK0d9gaImRWag0KT1ZhZoZF55vqYPFW50reJo7MMfDkCNjnb1EOWF9Mbx6UQeQP2CnRKofI</recordid><startdate>20020901</startdate><enddate>20020901</enddate><creator>MAHAPATRA, Nitish R</creator><creator>GHOSH, Sajalendu</creator><creator>DEB, Chirajyoti</creator><creator>BANERJEE, Pataki C</creator><general>Springer</general><general>Springer Nature B.V</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>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20020901</creationdate><title>Resistance to cadmium and zinc in Acidiphilium symbioticum KM2 is plasmid mediated</title><author>MAHAPATRA, Nitish R ; GHOSH, Sajalendu ; DEB, Chirajyoti ; BANERJEE, Pataki C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c419t-25513da528a83113e505bfb7ef7dd0d4dde95e4ff61296fe6273c366a4bdccd03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Acetobacteraceae - drug effects</topic><topic>Acetobacteraceae - genetics</topic><topic>Acidiphilium</topic><topic>Action of physical and chemical agents on bacteria</topic><topic>Amino Acid Sequence</topic><topic>Amino acids</topic><topic>Bacteriology</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cadmium</topic><topic>Cadmium - pharmacology</topic><topic>Cloning, Molecular</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Bacterial - genetics</topic><topic>Drug Resistance, Bacterial - genetics</topic><topic>E coli</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. 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The bacterium becomes extremely sensitive to metals when it is cured of its plasmids. A mini-plasmid library was constructed by ligating the plasmid DNA fragments generated by MboI partial digestion into the BamHI site of pBluescriptII KS+. The Lac(-)Amp(r) transformants of Escherichia coli DH5alpha, isolated after transformation with the library, were counter-selected on Cu(2+), Cd(2+), Ni(2+), and Zn(2+)-containing plates. Only Cd(2+)- and Zn(2+)-resistant colonies were developed, and, after screening, four types of recombinant plasmids designated as pNM201 (7.2 kb), pNM206 (3.4 kb), pNM208 (4.5 kb), and pNM215 (4.9 kb) were obtained. The DNA insert in pNM206 hybridized strongly with the 3.8-kb plasmid and weakly with the 7.1-kb plasmid of Acidiphilium symbioticum KM2. The DNA insert in pNM215 hybridized only with the 7.1-kb plasmid. These results strongly suggested that resistance to cadmium and zinc in A. symbioticum KM2 is mediated by these plasmids. The smallest insert of 422 bp in pNM206 conferring metal resistance in E. coli has no sequence similarity with the reported metal-resistant genes. All the putative ORFs are significantly rich (up to 37%) in basic amino acids, mainly arginine.</abstract><cop>New York, NY</cop><pub>Springer</pub><pmid>12177739</pmid><doi>10.1007/s00284-001-0113-6</doi><tpages>7</tpages></addata></record> |
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| subjects | Acetobacteraceae - drug effects Acetobacteraceae - genetics Acidiphilium Action of physical and chemical agents on bacteria Amino Acid Sequence Amino acids Bacteriology Base Sequence Biological and medical sciences Cadmium Cadmium - pharmacology Cloning, Molecular Deoxyribonucleic acid DNA DNA, Bacterial - genetics Drug Resistance, Bacterial - genetics E coli Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Genetics Metals Microbiology Mining Molecular Sequence Data Plasmids Plasmids - genetics Transformation, Genetic Zinc Zinc - pharmacology |
| title | Resistance to cadmium and zinc in Acidiphilium symbioticum KM2 is plasmid mediated |
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