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Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system
Bombyx mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system (BES) has a lot of advantages such as high expression efficiency, convenience, and low feeding cost. In this report, we used a recently developed BmNPV bacmid, which could infect both B. mori cell lines and silkworm larvae....
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| Published in: | Applied microbiology and biotechnology 2006-06, Vol.71 (2), p.192-199 |
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| Main Authors: | , , , , , , , , |
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| cited_by | cdi_FETCH-LOGICAL-c411t-6f15eef95ddb6fa899e35f79e250aa0366c36343bc0466ca58e8c451209810393 |
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| cites | cdi_FETCH-LOGICAL-c411t-6f15eef95ddb6fa899e35f79e250aa0366c36343bc0466ca58e8c451209810393 |
| container_end_page | 199 |
| container_issue | 2 |
| container_start_page | 192 |
| container_title | Applied microbiology and biotechnology |
| container_volume | 71 |
| creator | Miao, Y Zhang, Y Nakagaki, K Zhao, T Zhao, A Meng, Y Nakagaki, M Park, E.Y Maenaka, K |
| description | Bombyx mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system (BES) has a lot of advantages such as high expression efficiency, convenience, and low feeding cost. In this report, we used a recently developed BmNPV bacmid, which could infect both B. mori cell lines and silkworm larvae. The results showed it takes only 7 to 10 days to generate recombinant baculovirus and permit the rapid isolation from small-scale cultures and then use it to transfect B. mori cell lines, compared to traditional homologous recombination method, which needs at least 40 days for multiple rounds of purification and amplification of viruses. Using this BES, we expressed a recombinant spider flagelliform protein in BmN cell line, which was around 37 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. The BmNPV bacmid system using silkworm would be very attractive for expression of target proteins. |
| doi_str_mv | 10.1007/s00253-005-0127-2 |
| format | article |
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In this report, we used a recently developed BmNPV bacmid, which could infect both B. mori cell lines and silkworm larvae. The results showed it takes only 7 to 10 days to generate recombinant baculovirus and permit the rapid isolation from small-scale cultures and then use it to transfect B. mori cell lines, compared to traditional homologous recombination method, which needs at least 40 days for multiple rounds of purification and amplification of viruses. Using this BES, we expressed a recombinant spider flagelliform protein in BmN cell line, which was around 37 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. The BmNPV bacmid system using silkworm would be very attractive for expression of target proteins.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-005-0127-2</identifier><identifier>PMID: 16158284</identifier><identifier>CODEN: AMBIDG</identifier><language>eng</language><publisher>Berlin: Springer</publisher><subject>Amino Acid Sequence ; Animals ; Araneae ; Baculoviridae - genetics ; Baculovirus ; Base Sequence ; Biological and medical sciences ; Biotechnology ; Biotechnology - methods ; Blotting, Western ; Bombyx - metabolism ; Bombyx - virology ; Bombyx mori ; Bombyx mori NPV ; Cloning, Molecular ; DNA - chemistry ; DNA - genetics ; Enzymes ; Fundamental and applied biological sciences. Psychology ; Larvae ; Molecular Sequence Data ; Nuclear polyhedrosis virus ; Polymerase Chain Reaction ; Proteins ; Proteins - genetics ; Proteins - metabolism ; Recombinant Proteins - biosynthesis ; Recombinant Proteins - genetics ; Silk ; Spiders - genetics ; Spiders - metabolism</subject><ispartof>Applied microbiology and biotechnology, 2006-06, Vol.71 (2), p.192-199</ispartof><rights>2007 INIST-CNRS</rights><rights>Springer-Verlag 2006</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c411t-6f15eef95ddb6fa899e35f79e250aa0366c36343bc0466ca58e8c451209810393</citedby><cites>FETCH-LOGICAL-c411t-6f15eef95ddb6fa899e35f79e250aa0366c36343bc0466ca58e8c451209810393</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/625701205/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$H</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/625701205?pq-origsite=primo$$EHTML$$P50$$Gproquest$$H</linktohtml><link.rule.ids>314,780,784,11688,27924,27925,36060,36061,44363,74895</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17900961$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16158284$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Miao, Y</creatorcontrib><creatorcontrib>Zhang, Y</creatorcontrib><creatorcontrib>Nakagaki, K</creatorcontrib><creatorcontrib>Zhao, T</creatorcontrib><creatorcontrib>Zhao, A</creatorcontrib><creatorcontrib>Meng, Y</creatorcontrib><creatorcontrib>Nakagaki, M</creatorcontrib><creatorcontrib>Park, E.Y</creatorcontrib><creatorcontrib>Maenaka, K</creatorcontrib><title>Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><description>Bombyx mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system (BES) has a lot of advantages such as high expression efficiency, convenience, and low feeding cost. 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The BmNPV bacmid system using silkworm would be very attractive for expression of target proteins.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Araneae</subject><subject>Baculoviridae - genetics</subject><subject>Baculovirus</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Biotechnology - methods</subject><subject>Blotting, Western</subject><subject>Bombyx - metabolism</subject><subject>Bombyx - virology</subject><subject>Bombyx mori</subject><subject>Bombyx mori NPV</subject><subject>Cloning, Molecular</subject><subject>DNA - chemistry</subject><subject>DNA - genetics</subject><subject>Enzymes</subject><subject>Fundamental and applied biological sciences. 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Academic</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Miao, Y</au><au>Zhang, Y</au><au>Nakagaki, K</au><au>Zhao, T</au><au>Zhao, A</au><au>Meng, Y</au><au>Nakagaki, M</au><au>Park, E.Y</au><au>Maenaka, K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system</atitle><jtitle>Applied microbiology and biotechnology</jtitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2006-06-01</date><risdate>2006</risdate><volume>71</volume><issue>2</issue><spage>192</spage><epage>199</epage><pages>192-199</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><coden>AMBIDG</coden><abstract>Bombyx mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system (BES) has a lot of advantages such as high expression efficiency, convenience, and low feeding cost. In this report, we used a recently developed BmNPV bacmid, which could infect both B. mori cell lines and silkworm larvae. The results showed it takes only 7 to 10 days to generate recombinant baculovirus and permit the rapid isolation from small-scale cultures and then use it to transfect B. mori cell lines, compared to traditional homologous recombination method, which needs at least 40 days for multiple rounds of purification and amplification of viruses. Using this BES, we expressed a recombinant spider flagelliform protein in BmN cell line, which was around 37 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. The BmNPV bacmid system using silkworm would be very attractive for expression of target proteins.</abstract><cop>Berlin</cop><pub>Springer</pub><pmid>16158284</pmid><doi>10.1007/s00253-005-0127-2</doi><tpages>8</tpages></addata></record> |
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| ispartof | Applied microbiology and biotechnology, 2006-06, Vol.71 (2), p.192-199 |
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| language | eng |
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| source | ABI/INFORM Global; Springer Link |
| subjects | Amino Acid Sequence Animals Araneae Baculoviridae - genetics Baculovirus Base Sequence Biological and medical sciences Biotechnology Biotechnology - methods Blotting, Western Bombyx - metabolism Bombyx - virology Bombyx mori Bombyx mori NPV Cloning, Molecular DNA - chemistry DNA - genetics Enzymes Fundamental and applied biological sciences. Psychology Larvae Molecular Sequence Data Nuclear polyhedrosis virus Polymerase Chain Reaction Proteins Proteins - genetics Proteins - metabolism Recombinant Proteins - biosynthesis Recombinant Proteins - genetics Silk Spiders - genetics Spiders - metabolism |
| title | Expression of spider flagelliform silk protein in Bombyx mori cell line by a novel Bac-to-Bac/BmNPV baculovirus expression system |
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