BACULOVIRUS-BASED EXPRESSION OF AN INSECT VIRAL PROTEIN IN 12 DIFFERENT INSECT CELL LINES
The ability of 12 unique lepidopteran insect cell lines from Anticarsia gemmatalis, Heliothis virescens, Lymantria dispar (two lines), Mamestra brassica, Plutella xylostella, Spodoptera frugiperda (two lines), and Trichoplusia ni (three lines) to support production of a recombinant polydnavirus (PDV...
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| Veröffentlicht in: | In vitro cellular & developmental biology. Animal 2005-01, Vol.41 (1), p.43-49 |
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| creator | CHEN, Y. P GUNDERSEN-RINDAL, D. E LYNN, D. E |
| description | The ability of 12 unique lepidopteran insect cell lines from Anticarsia gemmatalis, Heliothis virescens, Lymantria dispar (two lines), Mamestra brassica, Plutella xylostella, Spodoptera frugiperda (two lines), and Trichoplusia ni (three lines) to support production of a recombinant polydnavirus (PDV) protein (GiPDV 1.1) expressed using the Bac-to-Bac™ baculovirus expression system was examined. Polydnavirus gene GiPDV 1.1 was cloned into the pFastBac baculovirus vector under the control of the polyhedron promoter, followed by generation of recombinant bacmid–GiPDV 1.1 by site-specific transposition. The ability of each insect cell line to support recombinant PDV gene expression was estimated using reverse transcriptase–polymerase chain reaction and Western blot. Each insect cell line infected with recombinant bacmid–GiPDV 1.1 and tested in this study was capable of supporting and producing recombinant protein. Time course expression analysis showed that 72–96 h after transfection to be the optimal time for harvest of recombinant protein for each insect cell line. |
| doi_str_mv | 10.1290/0412081.1 |
| format | Article |
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Each insect cell line infected with recombinant bacmid–GiPDV 1.1 and tested in this study was capable of supporting and producing recombinant protein. Time course expression analysis showed that 72–96 h after transfection to be the optimal time for harvest of recombinant protein for each insect cell line.</description><identifier>ISSN: 1071-2690</identifier><identifier>ISSN: 1543-706X</identifier><identifier>EISSN: 1543-706X</identifier><identifier>DOI: 10.1290/0412081.1</identifier><identifier>PMID: 15926859</identifier><identifier>CODEN: IVCAED</identifier><language>eng</language><publisher>Germany: Society for In Vitro Biology</publisher><subject>Animals ; Anticarsia gemmatalis ; bacmid ; Baculoviridae ; Baculoviridae - metabolism ; Baculovirus ; baculovirus expression vector ; Blotting, Western ; Brassica ; Cell culture techniques ; Cell Line ; Cell lines ; CELLULAR PATHOLOGY/VIROLOGY ; Cloning, Molecular ; DNA ; Electrophoresis, Polyacrylamide Gel ; gene expression ; gene transfer ; genetic vectors ; Genetic Vectors - genetics ; Glyptapanteles indiensis ; Heliothis virescens ; insect cell lines ; Insect genetics ; insect pests ; Insect proteins ; Insect vectors ; Insecta ; Lepidoptera ; Lepidoptera - cytology ; Lymantria dispar ; Mamestra ; Parasite hosts ; Parasitism ; Plutella xylostella ; Polydnaviridae ; Polydnaviridae - metabolism ; Polydnavirus ; recombinant protein ; recombinant proteins ; Reverse Transcriptase Polymerase Chain Reaction ; Spodoptera frugiperda ; Time Factors ; Transfection ; Trichoplusia ni ; viral proteins ; Viral Proteins - metabolism</subject><ispartof>In vitro cellular & developmental biology. 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P</creatorcontrib><creatorcontrib>GUNDERSEN-RINDAL, D. E</creatorcontrib><creatorcontrib>LYNN, D. E</creatorcontrib><title>BACULOVIRUS-BASED EXPRESSION OF AN INSECT VIRAL PROTEIN IN 12 DIFFERENT INSECT CELL LINES</title><title>In vitro cellular & developmental biology. Animal</title><addtitle>In Vitro Cell Dev Biol Anim</addtitle><description>The ability of 12 unique lepidopteran insect cell lines from Anticarsia gemmatalis, Heliothis virescens, Lymantria dispar (two lines), Mamestra brassica, Plutella xylostella, Spodoptera frugiperda (two lines), and Trichoplusia ni (three lines) to support production of a recombinant polydnavirus (PDV) protein (GiPDV 1.1) expressed using the Bac-to-Bac™ baculovirus expression system was examined. Polydnavirus gene GiPDV 1.1 was cloned into the pFastBac baculovirus vector under the control of the polyhedron promoter, followed by generation of recombinant bacmid–GiPDV 1.1 by site-specific transposition. The ability of each insect cell line to support recombinant PDV gene expression was estimated using reverse transcriptase–polymerase chain reaction and Western blot. Each insect cell line infected with recombinant bacmid–GiPDV 1.1 and tested in this study was capable of supporting and producing recombinant protein. Time course expression analysis showed that 72–96 h after transfection to be the optimal time for harvest of recombinant protein for each insect cell line.</description><subject>Animals</subject><subject>Anticarsia gemmatalis</subject><subject>bacmid</subject><subject>Baculoviridae</subject><subject>Baculoviridae - metabolism</subject><subject>Baculovirus</subject><subject>baculovirus expression vector</subject><subject>Blotting, Western</subject><subject>Brassica</subject><subject>Cell culture techniques</subject><subject>Cell Line</subject><subject>Cell lines</subject><subject>CELLULAR PATHOLOGY/VIROLOGY</subject><subject>Cloning, Molecular</subject><subject>DNA</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>gene expression</subject><subject>gene transfer</subject><subject>genetic vectors</subject><subject>Genetic Vectors - genetics</subject><subject>Glyptapanteles indiensis</subject><subject>Heliothis virescens</subject><subject>insect cell lines</subject><subject>Insect genetics</subject><subject>insect pests</subject><subject>Insect proteins</subject><subject>Insect vectors</subject><subject>Insecta</subject><subject>Lepidoptera</subject><subject>Lepidoptera - cytology</subject><subject>Lymantria dispar</subject><subject>Mamestra</subject><subject>Parasite hosts</subject><subject>Parasitism</subject><subject>Plutella xylostella</subject><subject>Polydnaviridae</subject><subject>Polydnaviridae - metabolism</subject><subject>Polydnavirus</subject><subject>recombinant protein</subject><subject>recombinant proteins</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Spodoptera frugiperda</subject><subject>Time Factors</subject><subject>Transfection</subject><subject>Trichoplusia ni</subject><subject>viral proteins</subject><subject>Viral Proteins - metabolism</subject><issn>1071-2690</issn><issn>1543-706X</issn><issn>1543-706X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqN0s1v0zAUAHALMbExduCOIOKwiUM2P3_FPmaZC5GiZGpaNE6W0zioVduMuD3w389VCkgcYL48y--nJ_s9I_QW8DUQhW8wA4IlXMMLdAac0TjB4uFl2OMEYiIUPkWvvV_hsBSIV-gUuCJCcnWGvt2m2byovubTeR3fprW-i_TD_VTXdV6VUTWJ0jLKy1pnsyiYtIjup9VM54fDCEh0l08meqrL2S-U6aKIirzU9Rt00tm1dxfHeI7mEz3LvsRF9TnP0iJumMS7GGgiXUulEiBpSxpg0DHipGNARdthbDvcka4RQBxVpMVkQUDYzoqWWrJg9BxdjnUfh_7H3vmd2Sz9wq3Xduv6vTcikQozSf4LQSWSc548Bwoi5XNgeJRSOMCPf8FVvx-2oS2GUKYSUFwF9GlEi6H3fnCdeRyWGzv8NIDNYc7mOGcDwb4_Ftw3G9f-kcfBBvBuBCu_64ffeUYU5_LQtA9jurO9sd-HpTfzmmCgGMIfCTGIq1E0y77fun_c5Ql3wrWa</recordid><startdate>200501</startdate><enddate>200501</enddate><creator>CHEN, Y. P</creator><creator>GUNDERSEN-RINDAL, D. E</creator><creator>LYNN, D. E</creator><general>Society for In Vitro Biology</general><scope>FBQ</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>3V.</scope><scope>4T-</scope><scope>7QL</scope><scope>7T7</scope><scope>7TK</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AF</scope><scope>8AO</scope><scope>8C1</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>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>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>S0X</scope><scope>7QO</scope><scope>7SS</scope><scope>7X8</scope></search><sort><creationdate>200501</creationdate><title>BACULOVIRUS-BASED EXPRESSION OF AN INSECT VIRAL PROTEIN IN 12 DIFFERENT INSECT CELL LINES</title><author>CHEN, Y. P ; GUNDERSEN-RINDAL, D. E ; LYNN, D. E</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b480t-1378ed3896183d2b141f42e8e4136df00af0f2fb612e392d02c216afa6d3a2c43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Animals</topic><topic>Anticarsia gemmatalis</topic><topic>bacmid</topic><topic>Baculoviridae</topic><topic>Baculoviridae - metabolism</topic><topic>Baculovirus</topic><topic>baculovirus expression vector</topic><topic>Blotting, Western</topic><topic>Brassica</topic><topic>Cell culture techniques</topic><topic>Cell Line</topic><topic>Cell lines</topic><topic>CELLULAR PATHOLOGY/VIROLOGY</topic><topic>Cloning, Molecular</topic><topic>DNA</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>gene expression</topic><topic>gene transfer</topic><topic>genetic vectors</topic><topic>Genetic Vectors - genetics</topic><topic>Glyptapanteles indiensis</topic><topic>Heliothis virescens</topic><topic>insect cell lines</topic><topic>Insect genetics</topic><topic>insect pests</topic><topic>Insect proteins</topic><topic>Insect vectors</topic><topic>Insecta</topic><topic>Lepidoptera</topic><topic>Lepidoptera - cytology</topic><topic>Lymantria dispar</topic><topic>Mamestra</topic><topic>Parasite hosts</topic><topic>Parasitism</topic><topic>Plutella xylostella</topic><topic>Polydnaviridae</topic><topic>Polydnaviridae - metabolism</topic><topic>Polydnavirus</topic><topic>recombinant protein</topic><topic>recombinant proteins</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Spodoptera frugiperda</topic><topic>Time Factors</topic><topic>Transfection</topic><topic>Trichoplusia ni</topic><topic>viral proteins</topic><topic>Viral Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>CHEN, Y. P</creatorcontrib><creatorcontrib>GUNDERSEN-RINDAL, D. E</creatorcontrib><creatorcontrib>LYNN, D. 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Animal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>CHEN, Y. P</au><au>GUNDERSEN-RINDAL, D. E</au><au>LYNN, D. E</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>BACULOVIRUS-BASED EXPRESSION OF AN INSECT VIRAL PROTEIN IN 12 DIFFERENT INSECT CELL LINES</atitle><jtitle>In vitro cellular & developmental biology. Animal</jtitle><addtitle>In Vitro Cell Dev Biol Anim</addtitle><date>2005-01</date><risdate>2005</risdate><volume>41</volume><issue>1</issue><spage>43</spage><epage>49</epage><pages>43-49</pages><issn>1071-2690</issn><issn>1543-706X</issn><eissn>1543-706X</eissn><coden>IVCAED</coden><abstract>The ability of 12 unique lepidopteran insect cell lines from Anticarsia gemmatalis, Heliothis virescens, Lymantria dispar (two lines), Mamestra brassica, Plutella xylostella, Spodoptera frugiperda (two lines), and Trichoplusia ni (three lines) to support production of a recombinant polydnavirus (PDV) protein (GiPDV 1.1) expressed using the Bac-to-Bac™ baculovirus expression system was examined. Polydnavirus gene GiPDV 1.1 was cloned into the pFastBac baculovirus vector under the control of the polyhedron promoter, followed by generation of recombinant bacmid–GiPDV 1.1 by site-specific transposition. The ability of each insect cell line to support recombinant PDV gene expression was estimated using reverse transcriptase–polymerase chain reaction and Western blot. Each insect cell line infected with recombinant bacmid–GiPDV 1.1 and tested in this study was capable of supporting and producing recombinant protein. Time course expression analysis showed that 72–96 h after transfection to be the optimal time for harvest of recombinant protein for each insect cell line.</abstract><cop>Germany</cop><pub>Society for In Vitro Biology</pub><pmid>15926859</pmid><doi>10.1290/0412081.1</doi><tpages>7</tpages></addata></record> |
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| ispartof | In vitro cellular & developmental biology. Animal, 2005-01, Vol.41 (1), p.43-49 |
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| subjects | Animals Anticarsia gemmatalis bacmid Baculoviridae Baculoviridae - metabolism Baculovirus baculovirus expression vector Blotting, Western Brassica Cell culture techniques Cell Line Cell lines CELLULAR PATHOLOGY/VIROLOGY Cloning, Molecular DNA Electrophoresis, Polyacrylamide Gel gene expression gene transfer genetic vectors Genetic Vectors - genetics Glyptapanteles indiensis Heliothis virescens insect cell lines Insect genetics insect pests Insect proteins Insect vectors Insecta Lepidoptera Lepidoptera - cytology Lymantria dispar Mamestra Parasite hosts Parasitism Plutella xylostella Polydnaviridae Polydnaviridae - metabolism Polydnavirus recombinant protein recombinant proteins Reverse Transcriptase Polymerase Chain Reaction Spodoptera frugiperda Time Factors Transfection Trichoplusia ni viral proteins Viral Proteins - metabolism |
| title | BACULOVIRUS-BASED EXPRESSION OF AN INSECT VIRAL PROTEIN IN 12 DIFFERENT INSECT CELL LINES |
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