Production of Fibrinolytic Enzyme in Plastid-Transformed Tobacco Plants
The earthworm fibrinolytic enzyme, which belongs to a group of serine proteases with strong fibrinolytic activity, has been used as an oral drug for prevention and treatment of thrombosis in East Asia. Fibrizyme is a fibrinolytic enzyme isolated from the earthworm Eisenia andrei. Here we report gene...
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| Veröffentlicht in: | Plant molecular biology reporter 2009-12, Vol.27 (4), p.448-453 |
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| creator | Ko, Suk-Min Yoo, Byung-Ho Lim, Jong-Min Oh, Kwang-Hoon Liu, Jae-Il Kim, Suk-Weon Liu, Jang-Ryol Choi, Kwan-Sam Yoon, Eui-Soo |
| description | The earthworm fibrinolytic enzyme, which belongs to a group of serine proteases with strong fibrinolytic activity, has been used as an oral drug for prevention and treatment of thrombosis in East Asia. Fibrizyme is a fibrinolytic enzyme isolated from the earthworm Eisenia andrei. Here we report genetic engineering of tobacco plastids with stable integration of the fibrizyme gene into the tobacco chloroplast genome. A plastid transformation vector was constructed by introducing various regulatory elements into fibrizyme cDNA. This vector was delivered by particle bombardment into tobacco leaf explants and plastid-transformed plants were subsequently regenerated into whole plants through several rounds of selection. We confirmed stable integration of the fibrizyme gene into the tobacco plastid genome by PCR and Southern blot analyses. Northern and Western blot analyses revealed that mRNA and protein of recombinant fibrizyme were highly expressed in transformed tobacco plants. |
| doi_str_mv | 10.1007/s11105-009-0113-4 |
| format | Article |
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Fibrizyme is a fibrinolytic enzyme isolated from the earthworm Eisenia andrei. Here we report genetic engineering of tobacco plastids with stable integration of the fibrizyme gene into the tobacco chloroplast genome. A plastid transformation vector was constructed by introducing various regulatory elements into fibrizyme cDNA. This vector was delivered by particle bombardment into tobacco leaf explants and plastid-transformed plants were subsequently regenerated into whole plants through several rounds of selection. We confirmed stable integration of the fibrizyme gene into the tobacco plastid genome by PCR and Southern blot analyses. Northern and Western blot analyses revealed that mRNA and protein of recombinant fibrizyme were highly expressed in transformed tobacco plants.</description><identifier>ISSN: 0735-9640</identifier><identifier>EISSN: 1572-9818</identifier><identifier>DOI: 10.1007/s11105-009-0113-4</identifier><language>eng</language><publisher>New York: New York : Springer-Verlag</publisher><subject>Bioinformatics ; Biomedical and Life Sciences ; Bombardment ; chloroplasts ; earthworms ; Eisenia andrei ; Enzymes ; Explants ; Expression vectors ; Fibrin ; fibrinolysis ; Genetic engineering ; Genetic transformation ; Genomes ; Integration ; leaves ; Life Sciences ; Metabolomics ; mRNA ; Particle bombardment ; Plant Breeding/Biotechnology ; Plant Sciences ; Plastids ; polymerase chain reaction ; Proteomics ; recombinant proteins ; Regulatory sequences ; Serine ; serine proteinases ; Southern blotting ; Thromboembolism ; Thrombosis ; Tobacco ; transgenic plants</subject><ispartof>Plant molecular biology reporter, 2009-12, Vol.27 (4), p.448-453</ispartof><rights>Springer-Verlag 2009</rights><rights>Plant Molecular Biology Reporter is a copyright of Springer, 2009.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c406t-fe078675b8fc3d405f099fccc35507a1460b981b1656460c0271ce1175afd7f3</citedby><cites>FETCH-LOGICAL-c406t-fe078675b8fc3d405f099fccc35507a1460b981b1656460c0271ce1175afd7f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s11105-009-0113-4$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s11105-009-0113-4$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>315,781,785,27926,27927,41490,42559,51321</link.rule.ids></links><search><creatorcontrib>Ko, Suk-Min</creatorcontrib><creatorcontrib>Yoo, Byung-Ho</creatorcontrib><creatorcontrib>Lim, Jong-Min</creatorcontrib><creatorcontrib>Oh, Kwang-Hoon</creatorcontrib><creatorcontrib>Liu, Jae-Il</creatorcontrib><creatorcontrib>Kim, Suk-Weon</creatorcontrib><creatorcontrib>Liu, Jang-Ryol</creatorcontrib><creatorcontrib>Choi, Kwan-Sam</creatorcontrib><creatorcontrib>Yoon, Eui-Soo</creatorcontrib><title>Production of Fibrinolytic Enzyme in Plastid-Transformed Tobacco Plants</title><title>Plant molecular biology reporter</title><addtitle>Plant Mol Biol Rep</addtitle><description>The earthworm fibrinolytic enzyme, which belongs to a group of serine proteases with strong fibrinolytic activity, has been used as an oral drug for prevention and treatment of thrombosis in East Asia. Fibrizyme is a fibrinolytic enzyme isolated from the earthworm Eisenia andrei. Here we report genetic engineering of tobacco plastids with stable integration of the fibrizyme gene into the tobacco chloroplast genome. A plastid transformation vector was constructed by introducing various regulatory elements into fibrizyme cDNA. This vector was delivered by particle bombardment into tobacco leaf explants and plastid-transformed plants were subsequently regenerated into whole plants through several rounds of selection. We confirmed stable integration of the fibrizyme gene into the tobacco plastid genome by PCR and Southern blot analyses. Northern and Western blot analyses revealed that mRNA and protein of recombinant fibrizyme were highly expressed in transformed tobacco plants.</description><subject>Bioinformatics</subject><subject>Biomedical and Life Sciences</subject><subject>Bombardment</subject><subject>chloroplasts</subject><subject>earthworms</subject><subject>Eisenia andrei</subject><subject>Enzymes</subject><subject>Explants</subject><subject>Expression vectors</subject><subject>Fibrin</subject><subject>fibrinolysis</subject><subject>Genetic engineering</subject><subject>Genetic transformation</subject><subject>Genomes</subject><subject>Integration</subject><subject>leaves</subject><subject>Life Sciences</subject><subject>Metabolomics</subject><subject>mRNA</subject><subject>Particle bombardment</subject><subject>Plant Breeding/Biotechnology</subject><subject>Plant Sciences</subject><subject>Plastids</subject><subject>polymerase chain reaction</subject><subject>Proteomics</subject><subject>recombinant proteins</subject><subject>Regulatory sequences</subject><subject>Serine</subject><subject>serine proteinases</subject><subject>Southern blotting</subject><subject>Thromboembolism</subject><subject>Thrombosis</subject><subject>Tobacco</subject><subject>transgenic plants</subject><issn>0735-9640</issn><issn>1572-9818</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kEFLwzAUx4MoOKcfwJMFz9H3miZpjzK2KQwcWM8hTZuRsTUz6Q7z05tRD5485UH-v_97_Ai5R3hCAPkcERE4BagoIDJaXJAJcpnTqsTykkxAMk4rUcA1uYlxC4mBspyQ5Tr49mgG5_vM22zhmuB6vzsNzmTz_vu07zLXZ-udjoNraR10H60P-67Nat9oY_z5rx_iLbmyehe7u993SurFvJ690tX78m32sqKmADFQ24EsheRNaQ1rC-AWqsoaYxjnIDUWApp0cYOCizQbyCWaDlFybVtp2ZQ8jrWH4L-OXRzU1h9DnzYqrLhI5WUpUgrHlAk-xtBZdQhur8NJIaizLjXqUkmXOutSRWLykYkp22-68Kf5H-hhhKz2Sm-Ci-rzIwdkgKLKGcvZD6KVdUM</recordid><startdate>20091201</startdate><enddate>20091201</enddate><creator>Ko, Suk-Min</creator><creator>Yoo, Byung-Ho</creator><creator>Lim, Jong-Min</creator><creator>Oh, Kwang-Hoon</creator><creator>Liu, Jae-Il</creator><creator>Kim, Suk-Weon</creator><creator>Liu, Jang-Ryol</creator><creator>Choi, Kwan-Sam</creator><creator>Yoon, Eui-Soo</creator><general>New York : Springer-Verlag</general><general>Springer-Verlag</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7QR</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FK</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>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>LK8</scope><scope>M0K</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope></search><sort><creationdate>20091201</creationdate><title>Production of Fibrinolytic Enzyme in Plastid-Transformed Tobacco Plants</title><author>Ko, Suk-Min ; Yoo, Byung-Ho ; Lim, Jong-Min ; Oh, Kwang-Hoon ; Liu, Jae-Il ; Kim, Suk-Weon ; Liu, Jang-Ryol ; Choi, Kwan-Sam ; Yoon, Eui-Soo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c406t-fe078675b8fc3d405f099fccc35507a1460b981b1656460c0271ce1175afd7f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Bioinformatics</topic><topic>Biomedical and Life Sciences</topic><topic>Bombardment</topic><topic>chloroplasts</topic><topic>earthworms</topic><topic>Eisenia andrei</topic><topic>Enzymes</topic><topic>Explants</topic><topic>Expression vectors</topic><topic>Fibrin</topic><topic>fibrinolysis</topic><topic>Genetic engineering</topic><topic>Genetic transformation</topic><topic>Genomes</topic><topic>Integration</topic><topic>leaves</topic><topic>Life Sciences</topic><topic>Metabolomics</topic><topic>mRNA</topic><topic>Particle bombardment</topic><topic>Plant Breeding/Biotechnology</topic><topic>Plant Sciences</topic><topic>Plastids</topic><topic>polymerase chain reaction</topic><topic>Proteomics</topic><topic>recombinant proteins</topic><topic>Regulatory sequences</topic><topic>Serine</topic><topic>serine proteinases</topic><topic>Southern blotting</topic><topic>Thromboembolism</topic><topic>Thrombosis</topic><topic>Tobacco</topic><topic>transgenic plants</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ko, Suk-Min</creatorcontrib><creatorcontrib>Yoo, Byung-Ho</creatorcontrib><creatorcontrib>Lim, Jong-Min</creatorcontrib><creatorcontrib>Oh, Kwang-Hoon</creatorcontrib><creatorcontrib>Liu, Jae-Il</creatorcontrib><creatorcontrib>Kim, Suk-Weon</creatorcontrib><creatorcontrib>Liu, Jang-Ryol</creatorcontrib><creatorcontrib>Choi, Kwan-Sam</creatorcontrib><creatorcontrib>Yoon, Eui-Soo</creatorcontrib><collection>AGRIS</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Chemoreception 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>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>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>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</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><jtitle>Plant molecular biology reporter</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ko, Suk-Min</au><au>Yoo, Byung-Ho</au><au>Lim, Jong-Min</au><au>Oh, Kwang-Hoon</au><au>Liu, Jae-Il</au><au>Kim, Suk-Weon</au><au>Liu, Jang-Ryol</au><au>Choi, Kwan-Sam</au><au>Yoon, Eui-Soo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Production of Fibrinolytic Enzyme in Plastid-Transformed Tobacco Plants</atitle><jtitle>Plant molecular biology reporter</jtitle><stitle>Plant Mol Biol Rep</stitle><date>2009-12-01</date><risdate>2009</risdate><volume>27</volume><issue>4</issue><spage>448</spage><epage>453</epage><pages>448-453</pages><issn>0735-9640</issn><eissn>1572-9818</eissn><abstract>The earthworm fibrinolytic enzyme, which belongs to a group of serine proteases with strong fibrinolytic activity, has been used as an oral drug for prevention and treatment of thrombosis in East Asia. Fibrizyme is a fibrinolytic enzyme isolated from the earthworm Eisenia andrei. Here we report genetic engineering of tobacco plastids with stable integration of the fibrizyme gene into the tobacco chloroplast genome. A plastid transformation vector was constructed by introducing various regulatory elements into fibrizyme cDNA. This vector was delivered by particle bombardment into tobacco leaf explants and plastid-transformed plants were subsequently regenerated into whole plants through several rounds of selection. We confirmed stable integration of the fibrizyme gene into the tobacco plastid genome by PCR and Southern blot analyses. Northern and Western blot analyses revealed that mRNA and protein of recombinant fibrizyme were highly expressed in transformed tobacco plants.</abstract><cop>New York</cop><pub>New York : Springer-Verlag</pub><doi>10.1007/s11105-009-0113-4</doi><tpages>6</tpages></addata></record> |
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| subjects | Bioinformatics Biomedical and Life Sciences Bombardment chloroplasts earthworms Eisenia andrei Enzymes Explants Expression vectors Fibrin fibrinolysis Genetic engineering Genetic transformation Genomes Integration leaves Life Sciences Metabolomics mRNA Particle bombardment Plant Breeding/Biotechnology Plant Sciences Plastids polymerase chain reaction Proteomics recombinant proteins Regulatory sequences Serine serine proteinases Southern blotting Thromboembolism Thrombosis Tobacco transgenic plants |
| title | Production of Fibrinolytic Enzyme in Plastid-Transformed Tobacco Plants |
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