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Transgenic Brassica carinata as a vehicle for the production of recombinant proteins in seeds
Hirudin, a blood anticoagulant protein from leeches, and beta-glucuronidase were produced in Brassica carinata Braun (Ethiopian mustard) seeds using oleosin as a carrier. Cotyledonary petioles were infected with Agrobacterium strains containing oleosin-glucuronidase (pCGNOBPGUS-A) or oleosin-hirudin...
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| Published in: | Plant cell reports 1998-01, Vol.17 (3), p.195-200 |
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| Main Authors: | , , |
| Format: | Article |
| Language: | English |
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| container_end_page | 200 |
| container_issue | 3 |
| container_start_page | 195 |
| container_title | Plant cell reports |
| container_volume | 17 |
| creator | Chaudhary, S Parmenter, D.L Moloney, M.M |
| description | Hirudin, a blood anticoagulant protein from leeches, and beta-glucuronidase were produced in Brassica carinata Braun (Ethiopian mustard) seeds using oleosin as a carrier. Cotyledonary petioles were infected with Agrobacterium strains containing oleosin-glucuronidase (pCGNOBPGUS-A) or oleosin-hirudin (PCGN-OBHIRT) constructs. Polymerase chain reaction and neomycin phosphotransferase II enzyme assays confirmed the presence of the fusion genes in plants regenerating under selection. The fusion polypeptides were correctly expressed and targeted to the oil-bodies of the seeds with high fidelity (ca. 90%). Recombinant protein was purified from all other cellular protein by a simple flotation process and cleaved from oil-bodies using the endoprotease, Factor Xa. Hirudin activity was measured using a colorimetric thrombin inhibition assay and an activity in the range of 0.2-0.4 antithrombin units per milligram of oil-body protein was detected. B. carinata offers an attractive alternative for the production of recombinant proteins using oleosin technology. |
| doi_str_mv | 10.1007/s002990050377 |
| format | article |
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Cotyledonary petioles were infected with Agrobacterium strains containing oleosin-glucuronidase (pCGNOBPGUS-A) or oleosin-hirudin (PCGN-OBHIRT) constructs. Polymerase chain reaction and neomycin phosphotransferase II enzyme assays confirmed the presence of the fusion genes in plants regenerating under selection. The fusion polypeptides were correctly expressed and targeted to the oil-bodies of the seeds with high fidelity (ca. 90%). Recombinant protein was purified from all other cellular protein by a simple flotation process and cleaved from oil-bodies using the endoprotease, Factor Xa. Hirudin activity was measured using a colorimetric thrombin inhibition assay and an activity in the range of 0.2-0.4 antithrombin units per milligram of oil-body protein was detected. B. carinata offers an attractive alternative for the production of recombinant proteins using oleosin technology.</description><identifier>ISSN: 0721-7714</identifier><identifier>EISSN: 1432-203X</identifier><identifier>DOI: 10.1007/s002990050377</identifier><identifier>PMID: 30736499</identifier><identifier>CODEN: PCRPD8</identifier><language>eng</language><publisher>Berlin: Springer</publisher><subject>anticoagulants ; beta-glucuronidase ; Biological and medical sciences ; biosynthesis ; Biotechnology ; Brassica carinata ; cotyledons ; drugs ; Flotation ; Fundamental and applied biological sciences. Psychology ; gene expression ; gene transfer ; genes ; Genetic engineering ; Genetic technics ; hirudin ; Methods. Procedures. Technologies ; Modification of gene expression level ; petioles ; protein synthesis ; Proteins ; recombinant proteins ; Seeds ; Transgenic animals and transgenic plants ; Transgenic plants</subject><ispartof>Plant cell reports, 1998-01, Vol.17 (3), p.195-200</ispartof><rights>1998 INIST-CNRS</rights><rights>Springer-Verlag Berlin Heidelberg 1998</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c402t-75de0af26883f3acc17b48e1d40d891b4bd09fa46d1cdba7203e4b36d49ec863</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=2097163$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30736499$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chaudhary, S</creatorcontrib><creatorcontrib>Parmenter, D.L</creatorcontrib><creatorcontrib>Moloney, M.M</creatorcontrib><title>Transgenic Brassica carinata as a vehicle for the production of recombinant proteins in seeds</title><title>Plant cell reports</title><addtitle>Plant Cell Rep</addtitle><description>Hirudin, a blood anticoagulant protein from leeches, and beta-glucuronidase were produced in Brassica carinata Braun (Ethiopian mustard) seeds using oleosin as a carrier. Cotyledonary petioles were infected with Agrobacterium strains containing oleosin-glucuronidase (pCGNOBPGUS-A) or oleosin-hirudin (PCGN-OBHIRT) constructs. Polymerase chain reaction and neomycin phosphotransferase II enzyme assays confirmed the presence of the fusion genes in plants regenerating under selection. The fusion polypeptides were correctly expressed and targeted to the oil-bodies of the seeds with high fidelity (ca. 90%). Recombinant protein was purified from all other cellular protein by a simple flotation process and cleaved from oil-bodies using the endoprotease, Factor Xa. Hirudin activity was measured using a colorimetric thrombin inhibition assay and an activity in the range of 0.2-0.4 antithrombin units per milligram of oil-body protein was detected. B. carinata offers an attractive alternative for the production of recombinant proteins using oleosin technology.</description><subject>anticoagulants</subject><subject>beta-glucuronidase</subject><subject>Biological and medical sciences</subject><subject>biosynthesis</subject><subject>Biotechnology</subject><subject>Brassica carinata</subject><subject>cotyledons</subject><subject>drugs</subject><subject>Flotation</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>gene expression</subject><subject>gene transfer</subject><subject>genes</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>hirudin</subject><subject>Methods. Procedures. Technologies</subject><subject>Modification of gene expression level</subject><subject>petioles</subject><subject>protein synthesis</subject><subject>Proteins</subject><subject>recombinant proteins</subject><subject>Seeds</subject><subject>Transgenic animals and transgenic plants</subject><subject>Transgenic plants</subject><issn>0721-7714</issn><issn>1432-203X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><recordid>eNp90cuLFDEQB-AgijuOHr1qEBEvrZVH53HUxRcseHAEL9JU57GbpSe9Jt2C_70ZZlzQg6c61EdRVT9CHjN4xQD06wrArQXoQWh9h2yYFLzjIL7dJRvQnHVaM3lGHtR6DdCaWt0nZwK0UNLaDfm-K5jrZcjJ0bcFa00OqcOSMi5IsVKkP8NVclOgcS50uQr0psx-dUuaM50jLcHN-7HxvBw6S0i50pRpDcHXh-RexKmGR6e6Jbv373bnH7uLzx8-nb-56JwEvnS69wEwcmWMiAKdY3qUJjAvwRvLRjl6sBGl8sz5EXU7L8hRKC9tcEaJLXl5HNsW-LGGugz7VF2YJsxhXuvAObdgBMCBvvgvZaoX0Ou-wWf_wOt5LbldMRgjjbSq_XBLuiNyZa61hDjclLTH8mtgMBziGf6Kp_knp6HruA_-Vv_Jo4HnJ4DV4RRbOC7VW8fBaqZEY0-PLOI84GVp5OsXDkwAN0aBMOI3upef0A</recordid><startdate>19980101</startdate><enddate>19980101</enddate><creator>Chaudhary, S</creator><creator>Parmenter, D.L</creator><creator>Moloney, M.M</creator><general>Springer</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>IQODW</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope><scope>7QO</scope><scope>7X8</scope></search><sort><creationdate>19980101</creationdate><title>Transgenic Brassica carinata as a vehicle for the production of recombinant proteins in seeds</title><author>Chaudhary, S ; Parmenter, D.L ; Moloney, M.M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c402t-75de0af26883f3acc17b48e1d40d891b4bd09fa46d1cdba7203e4b36d49ec863</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>anticoagulants</topic><topic>beta-glucuronidase</topic><topic>Biological and medical sciences</topic><topic>biosynthesis</topic><topic>Biotechnology</topic><topic>Brassica carinata</topic><topic>cotyledons</topic><topic>drugs</topic><topic>Flotation</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>gene expression</topic><topic>gene transfer</topic><topic>genes</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>hirudin</topic><topic>Methods. Procedures. Technologies</topic><topic>Modification of gene expression level</topic><topic>petioles</topic><topic>protein synthesis</topic><topic>Proteins</topic><topic>recombinant proteins</topic><topic>Seeds</topic><topic>Transgenic animals and transgenic plants</topic><topic>Transgenic plants</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chaudhary, S</creatorcontrib><creatorcontrib>Parmenter, D.L</creatorcontrib><creatorcontrib>Moloney, M.M</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection (Proquest)</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>ProQuest Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Agriculture Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>PML(ProQuest Medical Library)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Plant cell reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chaudhary, S</au><au>Parmenter, D.L</au><au>Moloney, M.M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Transgenic Brassica carinata as a vehicle for the production of recombinant proteins in seeds</atitle><jtitle>Plant cell reports</jtitle><addtitle>Plant Cell Rep</addtitle><date>1998-01-01</date><risdate>1998</risdate><volume>17</volume><issue>3</issue><spage>195</spage><epage>200</epage><pages>195-200</pages><issn>0721-7714</issn><eissn>1432-203X</eissn><coden>PCRPD8</coden><abstract>Hirudin, a blood anticoagulant protein from leeches, and beta-glucuronidase were produced in Brassica carinata Braun (Ethiopian mustard) seeds using oleosin as a carrier. Cotyledonary petioles were infected with Agrobacterium strains containing oleosin-glucuronidase (pCGNOBPGUS-A) or oleosin-hirudin (PCGN-OBHIRT) constructs. Polymerase chain reaction and neomycin phosphotransferase II enzyme assays confirmed the presence of the fusion genes in plants regenerating under selection. The fusion polypeptides were correctly expressed and targeted to the oil-bodies of the seeds with high fidelity (ca. 90%). Recombinant protein was purified from all other cellular protein by a simple flotation process and cleaved from oil-bodies using the endoprotease, Factor Xa. Hirudin activity was measured using a colorimetric thrombin inhibition assay and an activity in the range of 0.2-0.4 antithrombin units per milligram of oil-body protein was detected. B. carinata offers an attractive alternative for the production of recombinant proteins using oleosin technology.</abstract><cop>Berlin</cop><pub>Springer</pub><pmid>30736499</pmid><doi>10.1007/s002990050377</doi><tpages>6</tpages></addata></record> |
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| identifier | ISSN: 0721-7714 |
| ispartof | Plant cell reports, 1998-01, Vol.17 (3), p.195-200 |
| issn | 0721-7714 1432-203X |
| language | eng |
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| subjects | anticoagulants beta-glucuronidase Biological and medical sciences biosynthesis Biotechnology Brassica carinata cotyledons drugs Flotation Fundamental and applied biological sciences. Psychology gene expression gene transfer genes Genetic engineering Genetic technics hirudin Methods. Procedures. Technologies Modification of gene expression level petioles protein synthesis Proteins recombinant proteins Seeds Transgenic animals and transgenic plants Transgenic plants |
| title | Transgenic Brassica carinata as a vehicle for the production of recombinant proteins in seeds |
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