Efficient suppression of the amber codon in E. coli in vitro translation system
An mRNA encoding the esterase from Alicyclobacillus acidocaldarius with catalytically essential serine codon (ACG) replaced by an amber (UAG) codon was used to study the suppression in in vitro translation system. Suppression of UAG by tRNA Ser(CUA) was monitored by determination of the full-length...
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| Veröffentlicht in: | FEBS letters 2005-04, Vol.579 (10), p.2156-2160 |
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| creator | Agafonov, Dmitry E. Huang, Yiwei Grote, Michael Sprinzl, Mathias |
| description | An mRNA encoding the esterase from
Alicyclobacillus acidocaldarius with catalytically essential serine codon (ACG) replaced by an amber (UAG) codon was used to study the suppression in in vitro translation system. Suppression of UAG by tRNA
Ser(CUA) was monitored by determination of the full-length and active esterase. It was shown that commonly used increase of suppressor tRNA concentration inhibits protein production and therefore limits suppression. In situ deactivation of release factor by specific antibodies leads to efficient suppression already at low suppressor tRNA concentration and allows an in vitro synthesis of fully active enzyme in high yield undistinguishable from wild-type protein. |
| doi_str_mv | 10.1016/j.febslet.2005.03.004 |
| format | Article |
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Alicyclobacillus acidocaldarius with catalytically essential serine codon (ACG) replaced by an amber (UAG) codon was used to study the suppression in in vitro translation system. Suppression of UAG by tRNA
Ser(CUA) was monitored by determination of the full-length and active esterase. It was shown that commonly used increase of suppressor tRNA concentration inhibits protein production and therefore limits suppression. In situ deactivation of release factor by specific antibodies leads to efficient suppression already at low suppressor tRNA concentration and allows an in vitro synthesis of fully active enzyme in high yield undistinguishable from wild-type protein.</description><subject>Alicyclobacillus acidocaldarius</subject><subject>Base Sequence</subject><subject>Codon, Terminator</subject><subject>DNA Primers</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Esterase</subject><subject>Genes, Suppressor</subject><subject>In vitro translation</subject><subject>Mutagenesis, Site-Directed</subject><subject>Protein Biosynthesis</subject><subject>Release factor 1</subject><subject>Suppression</subject><subject>Termination</subject><subject>tRNA</subject><issn>0014-5793</issn><issn>1873-3468</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc1OwzAQhC0EglJ4BFBO3BLWcdzEJwQopUhIHICz5Tob4So_xXaL-vbYaiWOcPLamhmvviHkikJGgc5uV1mLS9ehz3IAngHLAIojMqFVyVJWzKpjMgGgRcpLwc7IuXMrCPeKilNyRnlFKWPFhLzWbWu0wcEnbrNeW3TOjEMyton_xET1S7SJHpvwZIakzsLcmThujbdj4q0aXKd8tLid89hfkJNWdQ4vD-eUfMzr98dF-vL69Px4_5LqgosiFbnGtsx1zgVoaDhShTSnnBWMMWSsWdK2EoKViosGRVlgXJ6KUqhZWQlgU3Kzz13b8WuDzsveOI1dpwYcN04GKfBZSJsSvhdqOzpnsZVra3pld5KCjCTlSh5IykhSApOBZPBdHz7YLHtsfl0HdEGw2Au-TYe7_6XKef2Qv8VaYivAAViex6i7fRQGYluDVrpYicbGWNReNqP5Y9sfyDibRg</recordid><startdate>20050411</startdate><enddate>20050411</enddate><creator>Agafonov, Dmitry E.</creator><creator>Huang, Yiwei</creator><creator>Grote, Michael</creator><creator>Sprinzl, Mathias</creator><general>Elsevier B.V</general><scope>6I.</scope><scope>AAFTH</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>C1K</scope></search><sort><creationdate>20050411</creationdate><title>Efficient suppression of the amber codon in E. coli in vitro translation system</title><author>Agafonov, Dmitry E. ; Huang, Yiwei ; Grote, Michael ; Sprinzl, Mathias</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4594-92cef72c2590c0d5e1ae121534333e33db1f89937a59de974e00011979a678903</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Alicyclobacillus acidocaldarius</topic><topic>Base Sequence</topic><topic>Codon, Terminator</topic><topic>DNA Primers</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Esterase</topic><topic>Genes, Suppressor</topic><topic>In vitro translation</topic><topic>Mutagenesis, Site-Directed</topic><topic>Protein Biosynthesis</topic><topic>Release factor 1</topic><topic>Suppression</topic><topic>Termination</topic><topic>tRNA</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Agafonov, Dmitry E.</creatorcontrib><creatorcontrib>Huang, Yiwei</creatorcontrib><creatorcontrib>Grote, Michael</creatorcontrib><creatorcontrib>Sprinzl, Mathias</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</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>Environmental Sciences and Pollution Management</collection><jtitle>FEBS letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Agafonov, Dmitry E.</au><au>Huang, Yiwei</au><au>Grote, Michael</au><au>Sprinzl, Mathias</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficient suppression of the amber codon in E. coli in vitro translation system</atitle><jtitle>FEBS letters</jtitle><addtitle>FEBS Lett</addtitle><date>2005-04-11</date><risdate>2005</risdate><volume>579</volume><issue>10</issue><spage>2156</spage><epage>2160</epage><pages>2156-2160</pages><issn>0014-5793</issn><eissn>1873-3468</eissn><abstract>An mRNA encoding the esterase from
Alicyclobacillus acidocaldarius with catalytically essential serine codon (ACG) replaced by an amber (UAG) codon was used to study the suppression in in vitro translation system. Suppression of UAG by tRNA
Ser(CUA) was monitored by determination of the full-length and active esterase. It was shown that commonly used increase of suppressor tRNA concentration inhibits protein production and therefore limits suppression. In situ deactivation of release factor by specific antibodies leads to efficient suppression already at low suppressor tRNA concentration and allows an in vitro synthesis of fully active enzyme in high yield undistinguishable from wild-type protein.</abstract><cop>England</cop><pub>Elsevier B.V</pub><pmid>15811334</pmid><doi>10.1016/j.febslet.2005.03.004</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Wiley Free Content; ScienceDirect Journals (5 years ago - present); EZB-FREE-00999 freely available EZB journals; Wiley Online Library All Journals; Alma/SFX Local Collection |
| subjects | Alicyclobacillus acidocaldarius Base Sequence Codon, Terminator DNA Primers Escherichia coli Escherichia coli - genetics Esterase Genes, Suppressor In vitro translation Mutagenesis, Site-Directed Protein Biosynthesis Release factor 1 Suppression Termination tRNA |
| title | Efficient suppression of the amber codon in E. coli in vitro translation system |
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