Site-directed mutagenesis with Escherichia coli DNA polymerase III holoenzyme
Escherichia coli DNA polymerase III holoenzyme was used to synthesize double-stranded DNA from M 13 single-stranded DNA hybridized to a phosphorylated synthetic oligodeoxynucleotide containing a nucleotide substitution. The resulting DNA was transfected into E. coli JM101 without further treatment....
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| Veröffentlicht in: | Gene 1988-01, Vol.62 (1), p.135-139 |
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| container_end_page | 139 |
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| container_issue | 1 |
| container_start_page | 135 |
| container_title | Gene |
| container_volume | 62 |
| creator | Tsurusluta, Naoya Maki, Hisaji Korn, Laurence Jay |
| description | Escherichia coli DNA polymerase III holoenzyme was used to synthesize double-stranded DNA from M 13 single-stranded DNA hybridized to a phosphorylated synthetic oligodeoxynucleotide containing a nucleotide substitution. The resulting DNA was transfected into
E. coli JM101 without further treatment. Sequence analysis of randomly chosen phage clones revealed that the efficiency of mutagenesis was nearly 50%, which is the theoretical maximum. Treatment with DNA ligase after DNA synthesis was not necessary to obtain high efficiency of mutagenesis. Thus, use of DNA polymerase III holoenzyme provides a simple and efficient procedure for site-directed mutagenesis. |
| doi_str_mv | 10.1016/0378-1119(88)90587-2 |
| format | Article |
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E. coli JM101 without further treatment. Sequence analysis of randomly chosen phage clones revealed that the efficiency of mutagenesis was nearly 50%, which is the theoretical maximum. Treatment with DNA ligase after DNA synthesis was not necessary to obtain high efficiency of mutagenesis. Thus, use of DNA polymerase III holoenzyme provides a simple and efficient procedure for site-directed mutagenesis.</description><identifier>ISSN: 0378-1119</identifier><identifier>EISSN: 1879-0038</identifier><identifier>DOI: 10.1016/0378-1119(88)90587-2</identifier><identifier>PMID: 3286374</identifier><identifier>CODEN: GENED6</identifier><language>eng</language><publisher>Lausanne: Elsevier B.V</publisher><subject>Bacterial Proteins - metabolism ; Base Sequence ; Biological and medical sciences ; Biotechnology ; Coliphages - genetics ; DNA Polymerase III - metabolism ; DNA, Recombinant ; DNA, Viral - biosynthesis ; DNA, Viral - genetics ; DNA-Directed DNA Polymerase - metabolism ; Escherichia coli ; Escherichia coli - enzymology ; Escherichia coli - genetics ; Fundamental and applied biological sciences. Psychology ; Genetic technics ; Genetic Techniques ; in vitro DNA synthesis ; Methods. Procedures. Technologies ; Molecular and cellular biology ; Molecular genetics ; Molecular Sequence Data ; Mutagenesis. Repair ; Mutant screening ; Mutation ; phage M 13 ; phage M13 ; Prokaryotes ; Recombinant DNA ; S SB protein ; single-stranded DNA ; synthetic oligodeoxy- nucleotide</subject><ispartof>Gene, 1988-01, Vol.62 (1), p.135-139</ispartof><rights>1988</rights><rights>1988 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-f60499365939f3266e3c15b795422b1f2d82a5f2c5ee0a1fd7f05359f87655213</citedby><cites>FETCH-LOGICAL-c417t-f60499365939f3266e3c15b795422b1f2d82a5f2c5ee0a1fd7f05359f87655213</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0378-1119(88)90587-2$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7779212$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3286374$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tsurusluta, Naoya</creatorcontrib><creatorcontrib>Maki, Hisaji</creatorcontrib><creatorcontrib>Korn, Laurence Jay</creatorcontrib><title>Site-directed mutagenesis with Escherichia coli DNA polymerase III holoenzyme</title><title>Gene</title><addtitle>Gene</addtitle><description>Escherichia coli DNA polymerase III holoenzyme was used to synthesize double-stranded DNA from M 13 single-stranded DNA hybridized to a phosphorylated synthetic oligodeoxynucleotide containing a nucleotide substitution. The resulting DNA was transfected into
E. coli JM101 without further treatment. Sequence analysis of randomly chosen phage clones revealed that the efficiency of mutagenesis was nearly 50%, which is the theoretical maximum. Treatment with DNA ligase after DNA synthesis was not necessary to obtain high efficiency of mutagenesis. Thus, use of DNA polymerase III holoenzyme provides a simple and efficient procedure for site-directed mutagenesis.</description><subject>Bacterial Proteins - metabolism</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Coliphages - genetics</subject><subject>DNA Polymerase III - metabolism</subject><subject>DNA, Recombinant</subject><subject>DNA, Viral - biosynthesis</subject><subject>DNA, Viral - genetics</subject><subject>DNA-Directed DNA Polymerase - metabolism</subject><subject>Escherichia coli</subject><subject>Escherichia coli - enzymology</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic technics</subject><subject>Genetic Techniques</subject><subject>in vitro DNA synthesis</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis. Repair</subject><subject>Mutant screening</subject><subject>Mutation</subject><subject>phage M 13</subject><subject>phage M13</subject><subject>Prokaryotes</subject><subject>Recombinant DNA</subject><subject>S SB protein</subject><subject>single-stranded DNA</subject><subject>synthetic oligodeoxy- nucleotide</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMlKA0EQhhtRNC5voDAHET2M9jK9XQRxDUQ9qOem01NtWmaJ3RMlPr0TE3K0LgVVX_0UH0KHBJ8TTMQFZlLlhBB9qtSZxlzJnG6gAVFS5xgztYkGa2QH7ab0gfvinG6jbUaVYLIYoMeX0EFehgiugzKrZ519hwZSSNl36CbZbXITiMFNgs1cW4Xs5ukqm7bVvIZoE2TD4TCbtFULzU8_2kdb3lYJDlZ9D73d3b5eP-Sj5_vh9dUodwWRXe4FLrRmgmumPaNCAHOEj6XmBaVj4mmpqOWeOg6ALfGl9Jgzrr2Sov-fsD10ssydxvZzBqkzdUgOqso20M6SIZwUAgveg8USdLFNKYI30xhqG-eGYLOwaBaKzEKRUcr8WTS0Pzta5c_GNZTro5W2fn-82tvkbOWjbVxIa0xKqSlZxFwuMehdfAWIJrkAjYOlb1O24f8_fgHHAYxQ</recordid><startdate>19880101</startdate><enddate>19880101</enddate><creator>Tsurusluta, Naoya</creator><creator>Maki, Hisaji</creator><creator>Korn, Laurence Jay</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>7QL</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>19880101</creationdate><title>Site-directed mutagenesis with Escherichia coli DNA polymerase III holoenzyme</title><author>Tsurusluta, Naoya ; Maki, Hisaji ; Korn, Laurence Jay</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-f60499365939f3266e3c15b795422b1f2d82a5f2c5ee0a1fd7f05359f87655213</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>Bacterial Proteins - metabolism</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Coliphages - genetics</topic><topic>DNA Polymerase III - metabolism</topic><topic>DNA, Recombinant</topic><topic>DNA, Viral - biosynthesis</topic><topic>DNA, Viral - genetics</topic><topic>DNA-Directed DNA Polymerase - metabolism</topic><topic>Escherichia coli</topic><topic>Escherichia coli - enzymology</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetic technics</topic><topic>Genetic Techniques</topic><topic>in vitro DNA synthesis</topic><topic>Methods. Procedures. Technologies</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis. Repair</topic><topic>Mutant screening</topic><topic>Mutation</topic><topic>phage M 13</topic><topic>phage M13</topic><topic>Prokaryotes</topic><topic>Recombinant DNA</topic><topic>S SB protein</topic><topic>single-stranded DNA</topic><topic>synthetic oligodeoxy- nucleotide</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tsurusluta, Naoya</creatorcontrib><creatorcontrib>Maki, Hisaji</creatorcontrib><creatorcontrib>Korn, Laurence Jay</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids 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>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tsurusluta, Naoya</au><au>Maki, Hisaji</au><au>Korn, Laurence Jay</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Site-directed mutagenesis with Escherichia coli DNA polymerase III holoenzyme</atitle><jtitle>Gene</jtitle><addtitle>Gene</addtitle><date>1988-01-01</date><risdate>1988</risdate><volume>62</volume><issue>1</issue><spage>135</spage><epage>139</epage><pages>135-139</pages><issn>0378-1119</issn><eissn>1879-0038</eissn><coden>GENED6</coden><abstract>Escherichia coli DNA polymerase III holoenzyme was used to synthesize double-stranded DNA from M 13 single-stranded DNA hybridized to a phosphorylated synthetic oligodeoxynucleotide containing a nucleotide substitution. The resulting DNA was transfected into
E. coli JM101 without further treatment. Sequence analysis of randomly chosen phage clones revealed that the efficiency of mutagenesis was nearly 50%, which is the theoretical maximum. Treatment with DNA ligase after DNA synthesis was not necessary to obtain high efficiency of mutagenesis. Thus, use of DNA polymerase III holoenzyme provides a simple and efficient procedure for site-directed mutagenesis.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>3286374</pmid><doi>10.1016/0378-1119(88)90587-2</doi><tpages>5</tpages></addata></record> |
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| ispartof | Gene, 1988-01, Vol.62 (1), p.135-139 |
| issn | 0378-1119 1879-0038 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Bacterial Proteins - metabolism Base Sequence Biological and medical sciences Biotechnology Coliphages - genetics DNA Polymerase III - metabolism DNA, Recombinant DNA, Viral - biosynthesis DNA, Viral - genetics DNA-Directed DNA Polymerase - metabolism Escherichia coli Escherichia coli - enzymology Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Genetic technics Genetic Techniques in vitro DNA synthesis Methods. Procedures. Technologies Molecular and cellular biology Molecular genetics Molecular Sequence Data Mutagenesis. Repair Mutant screening Mutation phage M 13 phage M13 Prokaryotes Recombinant DNA S SB protein single-stranded DNA synthetic oligodeoxy- nucleotide |
| title | Site-directed mutagenesis with Escherichia coli DNA polymerase III holoenzyme |
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