Optimization of the synthesis of porcine somatotropin in Escherichia coli
We report on the influence of choice of promoter and RNA polymerase, 5'-untranslated regions and ribosome binding sites, codon usage, leader peptide coding sequences and poly A tail in the 3'-untranslated region on the synthesis of porcine somatotropin (PST) in Escherichia coli. A total of...
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| Veröffentlicht in: | Applied microbiology and biotechnology 1993-06, Vol.39 (3), p.324-328 |
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| container_title | Applied microbiology and biotechnology |
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| creator | Wang, H O'Mahony, D.J McConnell, D.J Qi, S.Z |
| description | We report on the influence of choice of promoter and RNA polymerase, 5'-untranslated regions and ribosome binding sites, codon usage, leader peptide coding sequences and poly A tail in the 3'-untranslated region on the synthesis of porcine somatotropin (PST) in Escherichia coli. A total of 12 different constructs were tested in this study for the production of porcine somatotropin (PST) in E. coli. Several factors have significant effects on PST synthesis. In the presence of a strong promoter and a strong ribosome binding site, the next most important factor seems to be the combination of sequences at the 5'-end of the mRNA including both the 5'-untranslated region and the start of the coding sequence. Codon usage in the 5'-coding sequence per se is not important in determining the level of PST synthesis where high level expression is achieved from a strong ribosome binding site. However, where low level synthesis of recombinant PST (rPST) is achieved, codon usage in the 5'-coding sequence is important in determining the level of PST synthesis. Leader sequences dramatically reduce the level of PST synthesis. The presence of a poly A tail in the 3'-untranslated region has no significant effect on PST synthesis. |
| doi_str_mv | 10.1007/BF00192086 |
| format | Article |
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A total of 12 different constructs were tested in this study for the production of porcine somatotropin (PST) in E. coli. Several factors have significant effects on PST synthesis. In the presence of a strong promoter and a strong ribosome binding site, the next most important factor seems to be the combination of sequences at the 5'-end of the mRNA including both the 5'-untranslated region and the start of the coding sequence. Codon usage in the 5'-coding sequence per se is not important in determining the level of PST synthesis where high level expression is achieved from a strong ribosome binding site. However, where low level synthesis of recombinant PST (rPST) is achieved, codon usage in the 5'-coding sequence is important in determining the level of PST synthesis. Leader sequences dramatically reduce the level of PST synthesis. The presence of a poly A tail in the 3'-untranslated region has no significant effect on PST synthesis.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/BF00192086</identifier><identifier>PMID: 7763714</identifier><identifier>CODEN: AMBIDG</identifier><language>eng</language><publisher>Berlin: Springer</publisher><subject>Animals ; Base Sequence ; Biological and medical sciences ; Biotechnology ; Cloning, Molecular ; Codon - genetics ; Escherichia coli ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genetic engineering ; Genetic technics ; Genetic Vectors ; Growth Hormone - biosynthesis ; Growth Hormone - genetics ; Methods. Procedures. Technologies ; Modification of gene expression level ; Molecular Sequence Data ; optimization ; Plasmids - genetics ; Protein Biosynthesis ; Protein Sorting Signals - genetics ; Recombinant Proteins - biosynthesis ; Recombinant Proteins - genetics ; somatotropin ; Swine ; synthesis</subject><ispartof>Applied microbiology and biotechnology, 1993-06, Vol.39 (3), p.324-328</ispartof><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c366t-4bd0e52760bcfa5e8eb96accb60926afb774d45936da1b3f20b715a4d98b8fd43</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4816389$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7763714$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, H</creatorcontrib><creatorcontrib>O'Mahony, D.J</creatorcontrib><creatorcontrib>McConnell, D.J</creatorcontrib><creatorcontrib>Qi, S.Z</creatorcontrib><title>Optimization of the synthesis of porcine somatotropin in Escherichia coli</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><description>We report on the influence of choice of promoter and RNA polymerase, 5'-untranslated regions and ribosome binding sites, codon usage, leader peptide coding sequences and poly A tail in the 3'-untranslated region on the synthesis of porcine somatotropin (PST) in Escherichia coli. A total of 12 different constructs were tested in this study for the production of porcine somatotropin (PST) in E. coli. Several factors have significant effects on PST synthesis. In the presence of a strong promoter and a strong ribosome binding site, the next most important factor seems to be the combination of sequences at the 5'-end of the mRNA including both the 5'-untranslated region and the start of the coding sequence. Codon usage in the 5'-coding sequence per se is not important in determining the level of PST synthesis where high level expression is achieved from a strong ribosome binding site. However, where low level synthesis of recombinant PST (rPST) is achieved, codon usage in the 5'-coding sequence is important in determining the level of PST synthesis. Leader sequences dramatically reduce the level of PST synthesis. The presence of a poly A tail in the 3'-untranslated region has no significant effect on PST synthesis.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cloning, Molecular</subject><subject>Codon - genetics</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Genetic Vectors</subject><subject>Growth Hormone - biosynthesis</subject><subject>Growth Hormone - genetics</subject><subject>Methods. Procedures. Technologies</subject><subject>Modification of gene expression level</subject><subject>Molecular Sequence Data</subject><subject>optimization</subject><subject>Plasmids - genetics</subject><subject>Protein Biosynthesis</subject><subject>Protein Sorting Signals - genetics</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Recombinant Proteins - genetics</subject><subject>somatotropin</subject><subject>Swine</subject><subject>synthesis</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0EFLwzAUB_Agis7pxbvYg3gQqkmTvqRHHU4HAw_quSRp4iJtU5PuMD-9nSt6FB48eO_He_BH6IzgG4Ixv72fY0yKDAvYQxPCaJZiIGwfTTDhecrzQhyh4xg_BpUJgEN0yDlQTtgELZ673jXuS_bOt4m3Sb8ySdy0Q4subgedD9q1w9A3svd98J1rk6Eeol6Z4PTKyUT72p2gAyvraE7HPkVv84fX2VO6fH5czO6WqaYAfcpUhU2eccBKW5kbYVQBUmsFuMhAWsU5q1heUKgkUdRmWHGSS1YVQglbMTpFV7u7XfCfaxP7snFRm7qWrfHrWPJcYABO_4UEgIqCbuH1DurgYwzGll1wjQybkuByG3D5F_CAz8era9WY6peOiQ77y3Evo5a1DbLVLv4yJsjP1ym62DErfSnfw0DeXjJMKCZMUCYE_QZWFouO</recordid><startdate>19930601</startdate><enddate>19930601</enddate><creator>Wang, H</creator><creator>O'Mahony, D.J</creator><creator>McConnell, D.J</creator><creator>Qi, S.Z</creator><general>Springer</general><scope>FBQ</scope><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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19930601</creationdate><title>Optimization of the synthesis of porcine somatotropin in Escherichia coli</title><author>Wang, H ; O'Mahony, D.J ; McConnell, D.J ; Qi, S.Z</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c366t-4bd0e52760bcfa5e8eb96accb60926afb774d45936da1b3f20b715a4d98b8fd43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cloning, Molecular</topic><topic>Codon - genetics</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Genetic Vectors</topic><topic>Growth Hormone - biosynthesis</topic><topic>Growth Hormone - genetics</topic><topic>Methods. Procedures. Technologies</topic><topic>Modification of gene expression level</topic><topic>Molecular Sequence Data</topic><topic>optimization</topic><topic>Plasmids - genetics</topic><topic>Protein Biosynthesis</topic><topic>Protein Sorting Signals - genetics</topic><topic>Recombinant Proteins - biosynthesis</topic><topic>Recombinant Proteins - genetics</topic><topic>somatotropin</topic><topic>Swine</topic><topic>synthesis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, H</creatorcontrib><creatorcontrib>O'Mahony, D.J</creatorcontrib><creatorcontrib>McConnell, D.J</creatorcontrib><creatorcontrib>Qi, S.Z</creatorcontrib><collection>AGRIS</collection><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>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, H</au><au>O'Mahony, D.J</au><au>McConnell, D.J</au><au>Qi, S.Z</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Optimization of the synthesis of porcine somatotropin in Escherichia coli</atitle><jtitle>Applied microbiology and biotechnology</jtitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>1993-06-01</date><risdate>1993</risdate><volume>39</volume><issue>3</issue><spage>324</spage><epage>328</epage><pages>324-328</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><coden>AMBIDG</coden><abstract>We report on the influence of choice of promoter and RNA polymerase, 5'-untranslated regions and ribosome binding sites, codon usage, leader peptide coding sequences and poly A tail in the 3'-untranslated region on the synthesis of porcine somatotropin (PST) in Escherichia coli. A total of 12 different constructs were tested in this study for the production of porcine somatotropin (PST) in E. coli. Several factors have significant effects on PST synthesis. In the presence of a strong promoter and a strong ribosome binding site, the next most important factor seems to be the combination of sequences at the 5'-end of the mRNA including both the 5'-untranslated region and the start of the coding sequence. Codon usage in the 5'-coding sequence per se is not important in determining the level of PST synthesis where high level expression is achieved from a strong ribosome binding site. However, where low level synthesis of recombinant PST (rPST) is achieved, codon usage in the 5'-coding sequence is important in determining the level of PST synthesis. Leader sequences dramatically reduce the level of PST synthesis. The presence of a poly A tail in the 3'-untranslated region has no significant effect on PST synthesis.</abstract><cop>Berlin</cop><pub>Springer</pub><pmid>7763714</pmid><doi>10.1007/BF00192086</doi><tpages>5</tpages></addata></record> |
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| subjects | Animals Base Sequence Biological and medical sciences Biotechnology Cloning, Molecular Codon - genetics Escherichia coli Escherichia coli - genetics Escherichia coli - metabolism Fundamental and applied biological sciences. Psychology Gene Expression Genetic engineering Genetic technics Genetic Vectors Growth Hormone - biosynthesis Growth Hormone - genetics Methods. Procedures. Technologies Modification of gene expression level Molecular Sequence Data optimization Plasmids - genetics Protein Biosynthesis Protein Sorting Signals - genetics Recombinant Proteins - biosynthesis Recombinant Proteins - genetics somatotropin Swine synthesis |
| title | Optimization of the synthesis of porcine somatotropin in Escherichia coli |
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