Alkali Denaturation of Covalently Closed Circular Duplex Deoxyribonucleic Acid
The alkaline pH range over which the double stranded circular DNA of the replicative form of the bacterial virus, ÏX174, is reversibly denatured has been determined in order to specify accurately the conditions for a preparative procedure for this DNA based on selective alkali denaturation. The den...
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| Veröffentlicht in: | The Journal of biological chemistry 1970-05, Vol.245 (10), p.2704-2708 |
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| container_title | The Journal of biological chemistry |
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| creator | Rush, M G Warner, R C |
| description | The alkaline pH range over which the double stranded circular DNA of the replicative form of the bacterial virus, ÏX174, is
reversibly denatured has been determined in order to specify accurately the conditions for a preparative procedure for this
DNA based on selective alkali denaturation. The denaturation remains entirely reversible to pH 12.6 at an ionic strength of
about 0.25 over a range in which the sedimentation coefficient, after an initial drop, more than doubles. The sharp transition
above pH 12.6 in which the denaturation becomes completely irreversible is accompanied by only a small further increase in
sedimentation coefficient. Sedimentation-pH titration curves have been determined in detail for the replicative form of ÏX174
(molecular weight 3.4 x 10 6 daltons) and for the circular DNA of a penicillinase plasmid of Staphylococcus aureus (molecular weight 16.5 x 10 6 daltons). The latter curve is displaced to a lower pH and the two phase nature of the curve in the range of increasing sedimentation
coefficient is markedly accentuated. |
| doi_str_mv | 10.1016/S0021-9258(18)63125-4 |
| format | Article |
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reversibly denatured has been determined in order to specify accurately the conditions for a preparative procedure for this
DNA based on selective alkali denaturation. The denaturation remains entirely reversible to pH 12.6 at an ionic strength of
about 0.25 over a range in which the sedimentation coefficient, after an initial drop, more than doubles. The sharp transition
above pH 12.6 in which the denaturation becomes completely irreversible is accompanied by only a small further increase in
sedimentation coefficient. Sedimentation-pH titration curves have been determined in detail for the replicative form of ÏX174
(molecular weight 3.4 x 10 6 daltons) and for the circular DNA of a penicillinase plasmid of Staphylococcus aureus (molecular weight 16.5 x 10 6 daltons). The latter curve is displaced to a lower pH and the two phase nature of the curve in the range of increasing sedimentation
coefficient is markedly accentuated.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(18)63125-4</identifier><identifier>PMID: 5445805</identifier><language>eng</language><publisher>United States: American Society for Biochemistry and Molecular Biology</publisher><subject>Centrifugation, Zonal ; Chemical Phenomena ; Chemistry ; Coliphages ; DNA, Bacterial ; DNA, Viral ; Hydrogen-Ion Concentration ; Nucleic Acid Denaturation ; Osmolar Concentration ; Penicillinase ; Staphylococcus</subject><ispartof>The Journal of biological chemistry, 1970-05, Vol.245 (10), p.2704-2708</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c379t-7b745d42fe8c725252cb9043a50602fc63c4f9d7c7e7d72002516922fb919e3e3</citedby><cites>FETCH-LOGICAL-c379t-7b745d42fe8c725252cb9043a50602fc63c4f9d7c7e7d72002516922fb919e3e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/5445805$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rush, M G</creatorcontrib><creatorcontrib>Warner, R C</creatorcontrib><title>Alkali Denaturation of Covalently Closed Circular Duplex Deoxyribonucleic Acid</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The alkaline pH range over which the double stranded circular DNA of the replicative form of the bacterial virus, ÏX174, is
reversibly denatured has been determined in order to specify accurately the conditions for a preparative procedure for this
DNA based on selective alkali denaturation. The denaturation remains entirely reversible to pH 12.6 at an ionic strength of
about 0.25 over a range in which the sedimentation coefficient, after an initial drop, more than doubles. The sharp transition
above pH 12.6 in which the denaturation becomes completely irreversible is accompanied by only a small further increase in
sedimentation coefficient. Sedimentation-pH titration curves have been determined in detail for the replicative form of ÏX174
(molecular weight 3.4 x 10 6 daltons) and for the circular DNA of a penicillinase plasmid of Staphylococcus aureus (molecular weight 16.5 x 10 6 daltons). The latter curve is displaced to a lower pH and the two phase nature of the curve in the range of increasing sedimentation
coefficient is markedly accentuated.</description><subject>Centrifugation, Zonal</subject><subject>Chemical Phenomena</subject><subject>Chemistry</subject><subject>Coliphages</subject><subject>DNA, Bacterial</subject><subject>DNA, Viral</subject><subject>Hydrogen-Ion Concentration</subject><subject>Nucleic Acid Denaturation</subject><subject>Osmolar Concentration</subject><subject>Penicillinase</subject><subject>Staphylococcus</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1970</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kMtOwzAQRS0EKuXxCUiRkBAsAn7GybJKeUkVLACJneU4E2pw42InQP-elFbMLGZx753RHIROCL4kmGRXTxhTkhZU5Ockv8gYoSLlO2hMcM5SJsjrLhr_W_bRQYzveChekBEaCc5FjsUYPUzch3Y2mUKruz7ozvo28U1S-i_toO3cKimdj1AnpQ2mdzok037p4GdI-J9VsJVve-PAmmRibH2E9hrtIhxv5yF6ubl-Lu_S2ePtfTmZpYbJoktlJbmoOW0gN5KKoU1VYM60wBmmjcmY4U1RSyNB1pIObwiSFZQ2VUEKYMAO0dlm7zL4zx5ipxY2GnBOt-D7qHKekZwJORjFxmiCjzFAo5bBLnRYKYLVmqP646jWkBTJ1R9HxYfcyfZAXy2g_k9twQ366Uaf27f5tw2gKuvNHBaKcrHeTeXwzy8bznj6</recordid><startdate>19700525</startdate><enddate>19700525</enddate><creator>Rush, M G</creator><creator>Warner, R C</creator><general>American Society for Biochemistry and Molecular Biology</general><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>7X8</scope></search><sort><creationdate>19700525</creationdate><title>Alkali Denaturation of Covalently Closed Circular Duplex Deoxyribonucleic Acid</title><author>Rush, M G ; Warner, R C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c379t-7b745d42fe8c725252cb9043a50602fc63c4f9d7c7e7d72002516922fb919e3e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1970</creationdate><topic>Centrifugation, Zonal</topic><topic>Chemical Phenomena</topic><topic>Chemistry</topic><topic>Coliphages</topic><topic>DNA, Bacterial</topic><topic>DNA, Viral</topic><topic>Hydrogen-Ion Concentration</topic><topic>Nucleic Acid Denaturation</topic><topic>Osmolar Concentration</topic><topic>Penicillinase</topic><topic>Staphylococcus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rush, M G</creatorcontrib><creatorcontrib>Warner, R C</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rush, M G</au><au>Warner, R C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Alkali Denaturation of Covalently Closed Circular Duplex Deoxyribonucleic Acid</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1970-05-25</date><risdate>1970</risdate><volume>245</volume><issue>10</issue><spage>2704</spage><epage>2708</epage><pages>2704-2708</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The alkaline pH range over which the double stranded circular DNA of the replicative form of the bacterial virus, ÏX174, is
reversibly denatured has been determined in order to specify accurately the conditions for a preparative procedure for this
DNA based on selective alkali denaturation. The denaturation remains entirely reversible to pH 12.6 at an ionic strength of
about 0.25 over a range in which the sedimentation coefficient, after an initial drop, more than doubles. The sharp transition
above pH 12.6 in which the denaturation becomes completely irreversible is accompanied by only a small further increase in
sedimentation coefficient. Sedimentation-pH titration curves have been determined in detail for the replicative form of ÏX174
(molecular weight 3.4 x 10 6 daltons) and for the circular DNA of a penicillinase plasmid of Staphylococcus aureus (molecular weight 16.5 x 10 6 daltons). The latter curve is displaced to a lower pH and the two phase nature of the curve in the range of increasing sedimentation
coefficient is markedly accentuated.</abstract><cop>United States</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>5445805</pmid><doi>10.1016/S0021-9258(18)63125-4</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1970-05, Vol.245 (10), p.2704-2708 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_84618357 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Centrifugation, Zonal Chemical Phenomena Chemistry Coliphages DNA, Bacterial DNA, Viral Hydrogen-Ion Concentration Nucleic Acid Denaturation Osmolar Concentration Penicillinase Staphylococcus |
| title | Alkali Denaturation of Covalently Closed Circular Duplex Deoxyribonucleic Acid |
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