Cloning and expression of the MspI restriction and modification genes
The genes for the MspI restriction (R) and modification enzymes (recognition sequence CCGG) have been cloned into Escherichia coli using the vector pBR322. Clones carrying both genes have been isolated from libraries prepared with EcoRI, HindIII and BamW. The smallest fragment that encodes both acti...
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| Veröffentlicht in: | Gene 1988-04, Vol.64 (1), p.1-8 |
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| creator | Nwankwo, Donald O. Wilson, Geoffrey G. |
| description | The genes for the
MspI restriction (R) and modification enzymes (recognition sequence CCGG) have been cloned into
Escherichia coli using the vector pBR322. Clones carrying both genes have been isolated from libraries prepared with
EcoRI,
HindIII and
BamW. The smallest fragment that encodes both activities is a 3.6-kb
HindIII fragment. Plasmids purified from the clones are fully resistant to digestion by
MspI indicating that the modification gene is functional in
E. coli. The clones remain sensitive to phage infection, however, indicating that the endonuclease is dysfunctional. When the R gene is brought under the control of the inducible leftward promoter from phage λ the level of endonuclease increases and the level of methylase decreases, suggesting that the genes are transcribed in opposite directions. |
| doi_str_mv | 10.1016/0378-1119(88)90475-1 |
| format | Article |
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MspI restriction (R) and modification enzymes (recognition sequence CCGG) have been cloned into
Escherichia coli using the vector pBR322. Clones carrying both genes have been isolated from libraries prepared with
EcoRI,
HindIII and
BamW. The smallest fragment that encodes both activities is a 3.6-kb
HindIII fragment. Plasmids purified from the clones are fully resistant to digestion by
MspI indicating that the modification gene is functional in
E. coli. The clones remain sensitive to phage infection, however, indicating that the endonuclease is dysfunctional. When the R gene is brought under the control of the inducible leftward promoter from phage λ the level of endonuclease increases and the level of methylase decreases, suggesting that the genes are transcribed in opposite directions.</description><identifier>ISSN: 0378-1119</identifier><identifier>EISSN: 1879-0038</identifier><identifier>DOI: 10.1016/0378-1119(88)90475-1</identifier><identifier>PMID: 2456254</identifier><identifier>CODEN: GENED6</identifier><language>eng</language><publisher>Lausanne: Elsevier B.V</publisher><subject>Bacteriology ; Biological and medical sciences ; Biotechnology ; Cloning, Molecular ; Deoxyribonuclease HpaII ; DNA (Cytosine-5-)-Methyltransferases - genetics ; DNA Restriction Enzymes - genetics ; DNA-Cytosine Methylases ; E. coli ; Escherichia coli ; Escherichia coli - genetics ; Fundamental and applied biological sciences. Psychology ; Genes ; Genes, Bacterial ; Genetic engineering ; Genetic technics ; Metabolism. Enzymes ; Methods. Procedures. Technologies ; methylase ; Microbiology ; Moraxella - enzymology ; Moraxella - genetics ; MspI endonuclease ; phage λ pL promoter ; plasmid pBR322 ; Recombinant DNA</subject><ispartof>Gene, 1988-04, Vol.64 (1), p.1-8</ispartof><rights>1988</rights><rights>1989 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-1f6da1fc51249823b469e7ecfc43de6e8bf0350d664e822b0e513aca5e5d64e13</citedby><cites>FETCH-LOGICAL-c417t-1f6da1fc51249823b469e7ecfc43de6e8bf0350d664e822b0e513aca5e5d64e13</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)90475-1$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,777,781,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=6997429$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2456254$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nwankwo, Donald O.</creatorcontrib><creatorcontrib>Wilson, Geoffrey G.</creatorcontrib><title>Cloning and expression of the MspI restriction and modification genes</title><title>Gene</title><addtitle>Gene</addtitle><description>The genes for the
MspI restriction (R) and modification enzymes (recognition sequence CCGG) have been cloned into
Escherichia coli using the vector pBR322. Clones carrying both genes have been isolated from libraries prepared with
EcoRI,
HindIII and
BamW. The smallest fragment that encodes both activities is a 3.6-kb
HindIII fragment. Plasmids purified from the clones are fully resistant to digestion by
MspI indicating that the modification gene is functional in
E. coli. The clones remain sensitive to phage infection, however, indicating that the endonuclease is dysfunctional. When the R gene is brought under the control of the inducible leftward promoter from phage λ the level of endonuclease increases and the level of methylase decreases, suggesting that the genes are transcribed in opposite directions.</description><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cloning, Molecular</subject><subject>Deoxyribonuclease HpaII</subject><subject>DNA (Cytosine-5-)-Methyltransferases - genetics</subject><subject>DNA Restriction Enzymes - genetics</subject><subject>DNA-Cytosine Methylases</subject><subject>E. coli</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Metabolism. Enzymes</subject><subject>Methods. Procedures. Technologies</subject><subject>methylase</subject><subject>Microbiology</subject><subject>Moraxella - enzymology</subject><subject>Moraxella - genetics</subject><subject>MspI endonuclease</subject><subject>phage λ pL promoter</subject><subject>plasmid pBR322</subject><subject>Recombinant DNA</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>eNqFkMtKAzEUhoMoWi9voDALEV2MJpPrbAQp3qDiRtchTU40Mp2pyVT07c20pUtNFoH_fPk5fAgdE3xJMBFXmEpVEkLqc6UuaswkL8kWGhEl6xJjqrbRaIPsof2UPnA-nFe7aLdiXFScjdDtuOna0L4VpnUFfM8jpBS6tuh80b9D8ZTmj0XO-hhsP-QDNutc8MGaZfAGLaRDtONNk-Bo_R6g17vbl_FDOXm-fxzfTErLiOxL4oUzxFtOKlarik6ZqEGC9ZZRBwLU1GPKsROCgaqqKQZOqLGGA3c5IvQAna1657H7XOS19CwkC01jWugWSUtFqaD4f5DwfKWQGWQr0MYupQhez2OYmfijCdaDZj041INDrZReatZD_8m6fzGdgdt8WnvN89P13CRrGh9Na0PaYKKuJavqjF2vMMjSvgJEnWyA1oILEWyvXRf-3uMXTVaYQA</recordid><startdate>19880415</startdate><enddate>19880415</enddate><creator>Nwankwo, Donald O.</creator><creator>Wilson, Geoffrey G.</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><scope>7X8</scope></search><sort><creationdate>19880415</creationdate><title>Cloning and expression of the MspI restriction and modification genes</title><author>Nwankwo, Donald O. ; Wilson, Geoffrey G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-1f6da1fc51249823b469e7ecfc43de6e8bf0350d664e822b0e513aca5e5d64e13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cloning, Molecular</topic><topic>Deoxyribonuclease HpaII</topic><topic>DNA (Cytosine-5-)-Methyltransferases - genetics</topic><topic>DNA Restriction Enzymes - genetics</topic><topic>DNA-Cytosine Methylases</topic><topic>E. coli</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes</topic><topic>Genes, Bacterial</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Metabolism. Enzymes</topic><topic>Methods. Procedures. Technologies</topic><topic>methylase</topic><topic>Microbiology</topic><topic>Moraxella - enzymology</topic><topic>Moraxella - genetics</topic><topic>MspI endonuclease</topic><topic>phage λ pL promoter</topic><topic>plasmid pBR322</topic><topic>Recombinant DNA</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nwankwo, Donald O.</creatorcontrib><creatorcontrib>Wilson, Geoffrey G.</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><collection>MEDLINE - Academic</collection><jtitle>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nwankwo, Donald O.</au><au>Wilson, Geoffrey G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning and expression of the MspI restriction and modification genes</atitle><jtitle>Gene</jtitle><addtitle>Gene</addtitle><date>1988-04-15</date><risdate>1988</risdate><volume>64</volume><issue>1</issue><spage>1</spage><epage>8</epage><pages>1-8</pages><issn>0378-1119</issn><eissn>1879-0038</eissn><coden>GENED6</coden><abstract>The genes for the
MspI restriction (R) and modification enzymes (recognition sequence CCGG) have been cloned into
Escherichia coli using the vector pBR322. Clones carrying both genes have been isolated from libraries prepared with
EcoRI,
HindIII and
BamW. The smallest fragment that encodes both activities is a 3.6-kb
HindIII fragment. Plasmids purified from the clones are fully resistant to digestion by
MspI indicating that the modification gene is functional in
E. coli. The clones remain sensitive to phage infection, however, indicating that the endonuclease is dysfunctional. When the R gene is brought under the control of the inducible leftward promoter from phage λ the level of endonuclease increases and the level of methylase decreases, suggesting that the genes are transcribed in opposite directions.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>2456254</pmid><doi>10.1016/0378-1119(88)90475-1</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Gene, 1988-04, Vol.64 (1), p.1-8 |
| issn | 0378-1119 1879-0038 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_78336301 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Bacteriology Biological and medical sciences Biotechnology Cloning, Molecular Deoxyribonuclease HpaII DNA (Cytosine-5-)-Methyltransferases - genetics DNA Restriction Enzymes - genetics DNA-Cytosine Methylases E. coli Escherichia coli Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Genes Genes, Bacterial Genetic engineering Genetic technics Metabolism. Enzymes Methods. Procedures. Technologies methylase Microbiology Moraxella - enzymology Moraxella - genetics MspI endonuclease phage λ pL promoter plasmid pBR322 Recombinant DNA |
| title | Cloning and expression of the MspI restriction and modification genes |
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