Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells
We have used site‐specific mutagenesis to change the anticodon of a Xenopus laevis tyrosine tRNA gene so that it would recognize ochre codons. This tRNA gene is expressed when amplified in monkey cells as part of a SV40 recombinant and efficiently suppresses termination at both the ochre codon separ...
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| Veröffentlicht in: | The EMBO journal 1984-11, Vol.3 (11), p.2445-2452 |
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| container_title | The EMBO journal |
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| creator | Laski, F.A. Belagaje, R. Hudziak, R.M. Capecchi, M.R. Norton, G.P. Palese, P. RajBhandary, U.L. Sharp, P.A. |
| description | We have used site‐specific mutagenesis to change the anticodon of a Xenopus laevis tyrosine tRNA gene so that it would recognize ochre codons. This tRNA gene is expressed when amplified in monkey cells as part of a SV40 recombinant and efficiently suppresses termination at both the ochre codon separating the adenovirus 2 hexon gene from a 23‐kd downstream gene and the ochre codon at the end of the NS1 gene of influenza virus A/Tex/1/68. Termination at an amber codon of a NS1 gene of another influenza virus strain was not suppressed by the (Su+) ochre gene suggesting that in mammalian cells amber codons are not recognized by ochre suppressor tRNAs. Finally, microinjection into mammalian cells of both (Su+) ochre tRNA genes and selectible genes containing ochre nonsense mutations gives rise to colonies under selective conditions. We conclude that it should be possible to isolate a wide assortment of mammalian cell lines with ochre suppressor activity. |
| doi_str_mv | 10.1002/j.1460-2075.1984.tb02154.x |
| format | Article |
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This tRNA gene is expressed when amplified in monkey cells as part of a SV40 recombinant and efficiently suppresses termination at both the ochre codon separating the adenovirus 2 hexon gene from a 23‐kd downstream gene and the ochre codon at the end of the NS1 gene of influenza virus A/Tex/1/68. Termination at an amber codon of a NS1 gene of another influenza virus strain was not suppressed by the (Su+) ochre gene suggesting that in mammalian cells amber codons are not recognized by ochre suppressor tRNAs. Finally, microinjection into mammalian cells of both (Su+) ochre tRNA genes and selectible genes containing ochre nonsense mutations gives rise to colonies under selective conditions. We conclude that it should be possible to isolate a wide assortment of mammalian cell lines with ochre suppressor activity.</description><identifier>ISSN: 0261-4189</identifier><identifier>EISSN: 1460-2075</identifier><identifier>DOI: 10.1002/j.1460-2075.1984.tb02154.x</identifier><identifier>PMID: 6096120</identifier><language>eng</language><publisher>England</publisher><subject>Amino Acid Sequence ; Animals ; Base Sequence ; Cell Line ; Chlorocebus aethiops ; Cloning, Molecular ; Codon ; DNA Restriction Enzymes ; Genes, Synthetic ; Kidney ; Mutation ; Nucleic Acid Conformation ; RNA, Transfer, Amino Acyl - genetics ; Simian virus 40 - genetics ; Suppression, Genetic ; Transcription, Genetic ; Xenopus ; Xenopus laevis</subject><ispartof>The EMBO journal, 1984-11, Vol.3 (11), p.2445-2452</ispartof><rights>1984 European Molecular Biology Organization</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4244-6170a34383f28490fd5d11339d7430033dab585aa1a38dab4a942a96b4aa1ff3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC557710/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC557710/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/6096120$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Laski, F.A.</creatorcontrib><creatorcontrib>Belagaje, R.</creatorcontrib><creatorcontrib>Hudziak, R.M.</creatorcontrib><creatorcontrib>Capecchi, M.R.</creatorcontrib><creatorcontrib>Norton, G.P.</creatorcontrib><creatorcontrib>Palese, P.</creatorcontrib><creatorcontrib>RajBhandary, U.L.</creatorcontrib><creatorcontrib>Sharp, P.A.</creatorcontrib><title>Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells</title><title>The EMBO journal</title><addtitle>EMBO J</addtitle><description>We have used site‐specific mutagenesis to change the anticodon of a Xenopus laevis tyrosine tRNA gene so that it would recognize ochre codons. This tRNA gene is expressed when amplified in monkey cells as part of a SV40 recombinant and efficiently suppresses termination at both the ochre codon separating the adenovirus 2 hexon gene from a 23‐kd downstream gene and the ochre codon at the end of the NS1 gene of influenza virus A/Tex/1/68. Termination at an amber codon of a NS1 gene of another influenza virus strain was not suppressed by the (Su+) ochre gene suggesting that in mammalian cells amber codons are not recognized by ochre suppressor tRNAs. Finally, microinjection into mammalian cells of both (Su+) ochre tRNA genes and selectible genes containing ochre nonsense mutations gives rise to colonies under selective conditions. We conclude that it should be possible to isolate a wide assortment of mammalian cell lines with ochre suppressor activity.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Cell Line</subject><subject>Chlorocebus aethiops</subject><subject>Cloning, Molecular</subject><subject>Codon</subject><subject>DNA Restriction Enzymes</subject><subject>Genes, Synthetic</subject><subject>Kidney</subject><subject>Mutation</subject><subject>Nucleic Acid Conformation</subject><subject>RNA, Transfer, Amino Acyl - genetics</subject><subject>Simian virus 40 - genetics</subject><subject>Suppression, Genetic</subject><subject>Transcription, Genetic</subject><subject>Xenopus</subject><subject>Xenopus laevis</subject><issn>0261-4189</issn><issn>1460-2075</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1984</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVUctOwzAQtBAIyuMTkCwO3BJ2YzsPJA4F8VQBCbhbbuJQV0lc7BTavyehVQVHTjva2dm1Zwg5QQgRIDqbhshjCCJIRIhZysN2DBEKHi62yGBDbZMBRDEGHNNsj-x7PwUAkSa4S3ZjyGKMYEBGr8umnWhvPLUlVQ21-cRp6uezmdPeW0fbl6chfdeN7tiC6sVP39iGmobWqq5VZTpZrqvKH5KdUlVeH63rAXm7uX67ugtGz7f3V8NRkPOI8yDGBBTjLGVllPIMykIUiIxlRcIZAGOFGotUKIWKpR3mKuORyuIOKCxLdkAuVmtn83Gti1w3rVOVnDlTK7eUVhn5l2nMRL7bTylEkiB0-tO13tmPufatrI3vP6AabedeIocYRCK6wfPVYO6s906XmxsIsk9CTmVvt-ztln0Scp2EXHTi49-v3EjX1nf8cMV_mUov_7FZXj9ePvxg9g1U-Zpp</recordid><startdate>198411</startdate><enddate>198411</enddate><creator>Laski, F.A.</creator><creator>Belagaje, R.</creator><creator>Hudziak, R.M.</creator><creator>Capecchi, M.R.</creator><creator>Norton, G.P.</creator><creator>Palese, P.</creator><creator>RajBhandary, U.L.</creator><creator>Sharp, P.A.</creator><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>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>198411</creationdate><title>Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells</title><author>Laski, F.A. ; Belagaje, R. ; Hudziak, R.M. ; Capecchi, M.R. ; Norton, G.P. ; Palese, P. ; RajBhandary, U.L. ; Sharp, P.A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4244-6170a34383f28490fd5d11339d7430033dab585aa1a38dab4a942a96b4aa1ff3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1984</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Cell Line</topic><topic>Chlorocebus aethiops</topic><topic>Cloning, Molecular</topic><topic>Codon</topic><topic>DNA Restriction Enzymes</topic><topic>Genes, Synthetic</topic><topic>Kidney</topic><topic>Mutation</topic><topic>Nucleic Acid Conformation</topic><topic>RNA, Transfer, Amino Acyl - genetics</topic><topic>Simian virus 40 - genetics</topic><topic>Suppression, Genetic</topic><topic>Transcription, Genetic</topic><topic>Xenopus</topic><topic>Xenopus laevis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Laski, F.A.</creatorcontrib><creatorcontrib>Belagaje, R.</creatorcontrib><creatorcontrib>Hudziak, R.M.</creatorcontrib><creatorcontrib>Capecchi, M.R.</creatorcontrib><creatorcontrib>Norton, G.P.</creatorcontrib><creatorcontrib>Palese, P.</creatorcontrib><creatorcontrib>RajBhandary, U.L.</creatorcontrib><creatorcontrib>Sharp, P.A.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The EMBO journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Laski, F.A.</au><au>Belagaje, R.</au><au>Hudziak, R.M.</au><au>Capecchi, M.R.</au><au>Norton, G.P.</au><au>Palese, P.</au><au>RajBhandary, U.L.</au><au>Sharp, P.A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells</atitle><jtitle>The EMBO journal</jtitle><addtitle>EMBO J</addtitle><date>1984-11</date><risdate>1984</risdate><volume>3</volume><issue>11</issue><spage>2445</spage><epage>2452</epage><pages>2445-2452</pages><issn>0261-4189</issn><eissn>1460-2075</eissn><abstract>We have used site‐specific mutagenesis to change the anticodon of a Xenopus laevis tyrosine tRNA gene so that it would recognize ochre codons. This tRNA gene is expressed when amplified in monkey cells as part of a SV40 recombinant and efficiently suppresses termination at both the ochre codon separating the adenovirus 2 hexon gene from a 23‐kd downstream gene and the ochre codon at the end of the NS1 gene of influenza virus A/Tex/1/68. Termination at an amber codon of a NS1 gene of another influenza virus strain was not suppressed by the (Su+) ochre gene suggesting that in mammalian cells amber codons are not recognized by ochre suppressor tRNAs. Finally, microinjection into mammalian cells of both (Su+) ochre tRNA genes and selectible genes containing ochre nonsense mutations gives rise to colonies under selective conditions. We conclude that it should be possible to isolate a wide assortment of mammalian cell lines with ochre suppressor activity.</abstract><cop>England</cop><pmid>6096120</pmid><doi>10.1002/j.1460-2075.1984.tb02154.x</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | Amino Acid Sequence Animals Base Sequence Cell Line Chlorocebus aethiops Cloning, Molecular Codon DNA Restriction Enzymes Genes, Synthetic Kidney Mutation Nucleic Acid Conformation RNA, Transfer, Amino Acyl - genetics Simian virus 40 - genetics Suppression, Genetic Transcription, Genetic Xenopus Xenopus laevis |
| title | Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells |
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