Cloning, co-expression with an amidating enzyme, and activity of the scorpion toxin BmK ITa1 cDNA in insect cells
BmK ITa1 is an insect-specific neurotoxin from the Chinese scorpion Buthus martensi Karsch (Bmk). We succeeded in obtaining biologically active recombinant BmK ITa1 protein by simultaneous expression in insect cells of BmK ITa1 cDNA with an amidating enzyme expressed by the rat peptidylglycine alpha...
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| Veröffentlicht in: | Molecular biotechnology 2003-05, Vol.24 (1), p.21-26 |
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| creator | Liu, Zhen Yang, Guanzhen Li, Boliang Chi, Chengwu Wu, Xiangfu |
| description | BmK ITa1 is an insect-specific neurotoxin from the Chinese scorpion Buthus martensi Karsch (Bmk). We succeeded in obtaining biologically active recombinant BmK ITa1 protein by simultaneous expression in insect cells of BmK ITa1 cDNA with an amidating enzyme expressed by the rat peptidylglycine alpha-amidating monooxygenase (PAM) gene. We investigated the insecticidal efficacy of recombinant BmK ITa1/W (without coexpression of PAM), and of BmK ITa1/A (with coexpression of PAM) in 5th instar Bombyx mori, by injecting these recombinant toxins into larvae. The lethal time for 50% of larvae (LT50) was 9 h for BmK ITa1/A and 17 h for BmK ITa1/W. At 19 h after injection all of the larvae exposed to BmK ITa1/A had been killed, whereas only half of the larvae exposed to BmK ITa1/W had been killed. These results show that the simultaneous expression of an amidating enzyme can result in apparently higher insecticidal activity of BmK ITa1. |
| doi_str_mv | 10.1385/MB:24:1:21 |
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We succeeded in obtaining biologically active recombinant BmK ITa1 protein by simultaneous expression in insect cells of BmK ITa1 cDNA with an amidating enzyme expressed by the rat peptidylglycine alpha-amidating monooxygenase (PAM) gene. We investigated the insecticidal efficacy of recombinant BmK ITa1/W (without coexpression of PAM), and of BmK ITa1/A (with coexpression of PAM) in 5th instar Bombyx mori, by injecting these recombinant toxins into larvae. The lethal time for 50% of larvae (LT50) was 9 h for BmK ITa1/A and 17 h for BmK ITa1/W. At 19 h after injection all of the larvae exposed to BmK ITa1/A had been killed, whereas only half of the larvae exposed to BmK ITa1/W had been killed. These results show that the simultaneous expression of an amidating enzyme can result in apparently higher insecticidal activity of BmK ITa1.</description><identifier>ISSN: 1073-6085</identifier><identifier>EISSN: 1559-0305</identifier><identifier>EISSN: 1073-6085</identifier><identifier>DOI: 10.1385/MB:24:1:21</identifier><identifier>PMID: 12721493</identifier><identifier>CODEN: MLBOEO</identifier><language>eng</language><publisher>Totowa, NJ: Humana Press</publisher><subject>Animals ; Baculoviridae - genetics ; Baculoviridae - metabolism ; Biological and medical sciences ; Biotechnology ; Bombyx - drug effects ; Bombyx mori ; Buthus martensii ; Cloning ; Cloning, Molecular ; Dose-Response Relationship, Drug ; Enzymes ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation - genetics ; Insects ; Larva - drug effects ; Larvae ; Lethal Dose 50 ; Methods. Procedures. Technologies ; Mixed Function Oxygenases - biosynthesis ; Mixed Function Oxygenases - genetics ; Multienzyme Complexes - biosynthesis ; Multienzyme Complexes - genetics ; Neurotoxins ; Protein engineering ; Recombinant Proteins - biosynthesis ; Recombinant Proteins - genetics ; Recombinant Proteins - isolation & purification ; Recombinant Proteins - pharmacology ; Scorpion Venoms - biosynthesis ; Scorpion Venoms - genetics ; Scorpion Venoms - isolation & purification ; Scorpion Venoms - pharmacology ; Toxins</subject><ispartof>Molecular biotechnology, 2003-05, Vol.24 (1), p.21-26</ispartof><rights>2003 INIST-CNRS</rights><rights>Humana Press Inc. 2003</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c403t-705e6dc137ec231000c375e7c014f203778fe0c87ca592b449eb8527211d36b63</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14855568$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12721493$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Liu, Zhen</creatorcontrib><creatorcontrib>Yang, Guanzhen</creatorcontrib><creatorcontrib>Li, Boliang</creatorcontrib><creatorcontrib>Chi, Chengwu</creatorcontrib><creatorcontrib>Wu, Xiangfu</creatorcontrib><title>Cloning, co-expression with an amidating enzyme, and activity of the scorpion toxin BmK ITa1 cDNA in insect cells</title><title>Molecular biotechnology</title><addtitle>Mol Biotechnol</addtitle><description>BmK ITa1 is an insect-specific neurotoxin from the Chinese scorpion Buthus martensi Karsch (Bmk). We succeeded in obtaining biologically active recombinant BmK ITa1 protein by simultaneous expression in insect cells of BmK ITa1 cDNA with an amidating enzyme expressed by the rat peptidylglycine alpha-amidating monooxygenase (PAM) gene. We investigated the insecticidal efficacy of recombinant BmK ITa1/W (without coexpression of PAM), and of BmK ITa1/A (with coexpression of PAM) in 5th instar Bombyx mori, by injecting these recombinant toxins into larvae. The lethal time for 50% of larvae (LT50) was 9 h for BmK ITa1/A and 17 h for BmK ITa1/W. At 19 h after injection all of the larvae exposed to BmK ITa1/A had been killed, whereas only half of the larvae exposed to BmK ITa1/W had been killed. These results show that the simultaneous expression of an amidating enzyme can result in apparently higher insecticidal activity of BmK ITa1.</description><subject>Animals</subject><subject>Baculoviridae - genetics</subject><subject>Baculoviridae - metabolism</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Bombyx - drug effects</subject><subject>Bombyx mori</subject><subject>Buthus martensii</subject><subject>Cloning</subject><subject>Cloning, Molecular</subject><subject>Dose-Response Relationship, Drug</subject><subject>Enzymes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation - genetics</subject><subject>Insects</subject><subject>Larva - drug effects</subject><subject>Larvae</subject><subject>Lethal Dose 50</subject><subject>Methods. Procedures. Technologies</subject><subject>Mixed Function Oxygenases - biosynthesis</subject><subject>Mixed Function Oxygenases - genetics</subject><subject>Multienzyme Complexes - biosynthesis</subject><subject>Multienzyme Complexes - genetics</subject><subject>Neurotoxins</subject><subject>Protein engineering</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - isolation & purification</subject><subject>Recombinant Proteins - pharmacology</subject><subject>Scorpion Venoms - biosynthesis</subject><subject>Scorpion Venoms - genetics</subject><subject>Scorpion Venoms - isolation & purification</subject><subject>Scorpion Venoms - pharmacology</subject><subject>Toxins</subject><issn>1073-6085</issn><issn>1559-0305</issn><issn>1073-6085</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp90V9rFDEQAPAgiq3VFz-ABEEL0tXJv0323nrXVoutvtTnkMtmbcpuck1ytuenN0cPCn3waYbhN8MMg9BbAp8JU-LL5XxG-YzMKHmG9okQXQMMxPOag2RNC0rsoVc53wBQIjh7ifYIlZTwju2j28UYgw-_j7CNjbtfJZezjwHf-XKNTcBm8r0pFWAX_m4md1SLPTa2-D--bHAccLl2ONuYVtu2Eu99wPPpOz6_MgTbkx_HuBZ8yM4WbN045tfoxWDG7N7s4gH6dXZ6tfjWXPz8er44vmgsB1YaCcK1vSVMOksZAQDLpHDSAuEDBSalGhxYJa0RHV1y3rmlEtu7SM_aZcsO0OHD3FWKt2uXi5583m5ggovrrDvBhZSdgio__lcSpQAYpxW-fwJv4jqFeoWmrJVKVlPRpwdkU8w5uUGvkp9M2mgCevsvfTnXlGuiKan43W7iejm5_pHuHlTBhx0w2ZpxSCZYnx8dV0KIVrF_mjyaTg</recordid><startdate>20030501</startdate><enddate>20030501</enddate><creator>Liu, Zhen</creator><creator>Yang, Guanzhen</creator><creator>Li, Boliang</creator><creator>Chi, Chengwu</creator><creator>Wu, Xiangfu</creator><general>Humana Press</general><general>Springer Nature B.V</general><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>3V.</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</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>L6V</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>Q9U</scope><scope>7SS</scope><scope>RC3</scope><scope>7U7</scope></search><sort><creationdate>20030501</creationdate><title>Cloning, co-expression with an amidating enzyme, and activity of the scorpion toxin BmK ITa1 cDNA in insect cells</title><author>Liu, Zhen ; Yang, Guanzhen ; Li, Boliang ; Chi, Chengwu ; Wu, Xiangfu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c403t-705e6dc137ec231000c375e7c014f203778fe0c87ca592b449eb8527211d36b63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>Baculoviridae - genetics</topic><topic>Baculoviridae - metabolism</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Bombyx - drug effects</topic><topic>Bombyx mori</topic><topic>Buthus martensii</topic><topic>Cloning</topic><topic>Cloning, Molecular</topic><topic>Dose-Response Relationship, Drug</topic><topic>Enzymes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation - genetics</topic><topic>Insects</topic><topic>Larva - drug effects</topic><topic>Larvae</topic><topic>Lethal Dose 50</topic><topic>Methods. Procedures. Technologies</topic><topic>Mixed Function Oxygenases - biosynthesis</topic><topic>Mixed Function Oxygenases - genetics</topic><topic>Multienzyme Complexes - biosynthesis</topic><topic>Multienzyme Complexes - genetics</topic><topic>Neurotoxins</topic><topic>Protein engineering</topic><topic>Recombinant Proteins - biosynthesis</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - isolation & purification</topic><topic>Recombinant Proteins - pharmacology</topic><topic>Scorpion Venoms - biosynthesis</topic><topic>Scorpion Venoms - genetics</topic><topic>Scorpion Venoms - isolation & purification</topic><topic>Scorpion Venoms - pharmacology</topic><topic>Toxins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Liu, Zhen</creatorcontrib><creatorcontrib>Yang, Guanzhen</creatorcontrib><creatorcontrib>Li, Boliang</creatorcontrib><creatorcontrib>Chi, Chengwu</creatorcontrib><creatorcontrib>Wu, 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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>Engineering Collection</collection><collection>ProQuest Central Basic</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Genetics Abstracts</collection><collection>Toxicology Abstracts</collection><jtitle>Molecular biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Liu, Zhen</au><au>Yang, Guanzhen</au><au>Li, Boliang</au><au>Chi, Chengwu</au><au>Wu, Xiangfu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning, co-expression with an amidating enzyme, and activity of the scorpion toxin BmK ITa1 cDNA in insect cells</atitle><jtitle>Molecular biotechnology</jtitle><addtitle>Mol Biotechnol</addtitle><date>2003-05-01</date><risdate>2003</risdate><volume>24</volume><issue>1</issue><spage>21</spage><epage>26</epage><pages>21-26</pages><issn>1073-6085</issn><eissn>1559-0305</eissn><eissn>1073-6085</eissn><coden>MLBOEO</coden><abstract>BmK ITa1 is an insect-specific neurotoxin from the Chinese scorpion Buthus martensi Karsch (Bmk). We succeeded in obtaining biologically active recombinant BmK ITa1 protein by simultaneous expression in insect cells of BmK ITa1 cDNA with an amidating enzyme expressed by the rat peptidylglycine alpha-amidating monooxygenase (PAM) gene. We investigated the insecticidal efficacy of recombinant BmK ITa1/W (without coexpression of PAM), and of BmK ITa1/A (with coexpression of PAM) in 5th instar Bombyx mori, by injecting these recombinant toxins into larvae. The lethal time for 50% of larvae (LT50) was 9 h for BmK ITa1/A and 17 h for BmK ITa1/W. At 19 h after injection all of the larvae exposed to BmK ITa1/A had been killed, whereas only half of the larvae exposed to BmK ITa1/W had been killed. These results show that the simultaneous expression of an amidating enzyme can result in apparently higher insecticidal activity of BmK ITa1.</abstract><cop>Totowa, NJ</cop><pub>Humana Press</pub><pmid>12721493</pmid><doi>10.1385/MB:24:1:21</doi><tpages>6</tpages></addata></record> |
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| subjects | Animals Baculoviridae - genetics Baculoviridae - metabolism Biological and medical sciences Biotechnology Bombyx - drug effects Bombyx mori Buthus martensii Cloning Cloning, Molecular Dose-Response Relationship, Drug Enzymes Fundamental and applied biological sciences. Psychology Gene Expression Regulation - genetics Insects Larva - drug effects Larvae Lethal Dose 50 Methods. Procedures. Technologies Mixed Function Oxygenases - biosynthesis Mixed Function Oxygenases - genetics Multienzyme Complexes - biosynthesis Multienzyme Complexes - genetics Neurotoxins Protein engineering Recombinant Proteins - biosynthesis Recombinant Proteins - genetics Recombinant Proteins - isolation & purification Recombinant Proteins - pharmacology Scorpion Venoms - biosynthesis Scorpion Venoms - genetics Scorpion Venoms - isolation & purification Scorpion Venoms - pharmacology Toxins |
| title | Cloning, co-expression with an amidating enzyme, and activity of the scorpion toxin BmK ITa1 cDNA in insect cells |
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