Increased toxicity of modified mosquitocidal binary toxins of Bacillus sphaericus expressed in Escherichia coli
The binary mosquitocidal genes of 51-kDa and 42-kDa proteins isolated from Bacillus sphaericus 1593 have been expressed at moderate levels in Escherichia coli employing the pQE expression system. The expressed proteins are readily visible in Coomassie-blue-stained protein gels. The recombinant E. co...
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| Veröffentlicht in: | Applied microbiology and biotechnology 1998-02, Vol.49 (2), p.164-167 |
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| creator | Ahmad, S Selvapandiyan, a Bhatnagar, R.K |
| description | The binary mosquitocidal genes of 51-kDa and 42-kDa proteins isolated from Bacillus sphaericus 1593 have been expressed at moderate levels in Escherichia coli employing the pQE expression system. The expressed proteins are readily visible in Coomassie-blue-stained protein gels. The recombinant E. coli cells expressing toxic proteins were toxic towards Culex larvae. During the assembly of crystals in B. sphaericus, the 42-kDa toxin is first cleaved at the N-terminal end by a specific B. sphaericus protease. To express the toxins in E. coli the B. sphaericus specific protease-recognition site was deleted at the N-terminal end of the 42-kDa toxin, thereby mimicking the structure of the toxin as present in the crystal. This modification resulted in a twofold increase in the toxicity of the E. coli cells expressing the modified 42-kDa toxin as a constituent of the binary toxin. Our results demonstrate the utility of this modification for heterologous expression of the binary toxin genes from B. sphaericus. |
| doi_str_mv | 10.1007/s002530051153 |
| format | Article |
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The expressed proteins are readily visible in Coomassie-blue-stained protein gels. The recombinant E. coli cells expressing toxic proteins were toxic towards Culex larvae. During the assembly of crystals in B. sphaericus, the 42-kDa toxin is first cleaved at the N-terminal end by a specific B. sphaericus protease. To express the toxins in E. coli the B. sphaericus specific protease-recognition site was deleted at the N-terminal end of the 42-kDa toxin, thereby mimicking the structure of the toxin as present in the crystal. This modification resulted in a twofold increase in the toxicity of the E. coli cells expressing the modified 42-kDa toxin as a constituent of the binary toxin. Our results demonstrate the utility of this modification for heterologous expression of the binary toxin genes from B. sphaericus.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s002530051153</identifier><identifier>PMID: 9534256</identifier><identifier>CODEN: AMBIDG</identifier><language>eng</language><publisher>Berlin: Springer</publisher><subject>animal health ; animal parasites and pests ; Animals ; Aquatic insects ; arthropod pests ; Bacillus - genetics ; Bacteria ; Bacterial Toxins - biosynthesis ; Bacterial Toxins - genetics ; Bacterial Toxins - toxicity ; Biological and medical sciences ; Biological Sciences ; Biotechnology ; Crystals ; Culex ; E coli ; Electrophoresis, Polyacrylamide Gel ; Enzymes ; Escherichia coli - genetics ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Genetic engineering ; Genetic technics ; Larva ; Larvae ; Methods. Procedures. Technologies ; Microbiology ; Modification of gene expression level ; Plasmids ; Proteins ; Recombinant Proteins - biosynthesis ; Recombinant Proteins - toxicity ; Toxicity ; Toxins</subject><ispartof>Applied microbiology and biotechnology, 1998-02, Vol.49 (2), p.164-167</ispartof><rights>1998 INIST-CNRS</rights><rights>Springer-Verlag Berlin Heidelberg 1998</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c399t-29e23a20b0edd672c2b896f60a5b60a1b2bf6c8d15c6fb1966e8dbd4496934de3</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=2170668$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9534256$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ahmad, S</creatorcontrib><creatorcontrib>Selvapandiyan, a</creatorcontrib><creatorcontrib>Bhatnagar, R.K</creatorcontrib><title>Increased toxicity of modified mosquitocidal binary toxins of Bacillus sphaericus expressed in Escherichia coli</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><description>The binary mosquitocidal genes of 51-kDa and 42-kDa proteins isolated from Bacillus sphaericus 1593 have been expressed at moderate levels in Escherichia coli employing the pQE expression system. The expressed proteins are readily visible in Coomassie-blue-stained protein gels. The recombinant E. coli cells expressing toxic proteins were toxic towards Culex larvae. During the assembly of crystals in B. sphaericus, the 42-kDa toxin is first cleaved at the N-terminal end by a specific B. sphaericus protease. To express the toxins in E. coli the B. sphaericus specific protease-recognition site was deleted at the N-terminal end of the 42-kDa toxin, thereby mimicking the structure of the toxin as present in the crystal. This modification resulted in a twofold increase in the toxicity of the E. coli cells expressing the modified 42-kDa toxin as a constituent of the binary toxin. Our results demonstrate the utility of this modification for heterologous expression of the binary toxin genes from B. sphaericus.</description><subject>animal health</subject><subject>animal parasites and pests</subject><subject>Animals</subject><subject>Aquatic insects</subject><subject>arthropod pests</subject><subject>Bacillus - genetics</subject><subject>Bacteria</subject><subject>Bacterial Toxins - biosynthesis</subject><subject>Bacterial Toxins - genetics</subject><subject>Bacterial Toxins - toxicity</subject><subject>Biological and medical sciences</subject><subject>Biological Sciences</subject><subject>Biotechnology</subject><subject>Crystals</subject><subject>Culex</subject><subject>E coli</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzymes</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Larva</subject><subject>Larvae</subject><subject>Methods. Procedures. Technologies</subject><subject>Microbiology</subject><subject>Modification of gene expression level</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Recombinant Proteins - toxicity</subject><subject>Toxicity</subject><subject>Toxins</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</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>eNpd0c1rFTEQAPAgSn1Wjx7FRcTbaj42s8lRS9VCwYP2vOTTl7K7ec3sQvvfm_U9CnrJhJkfwzBDyGtGPzJK-09IKZeCUsmYFE_IjnWCtxRY95TsKOtl20utnpMXiLeUMq4AzsiZlqLjEnYkX82uBIPBN0u-Ty4tD02OzZR9iqkmp4x3a1qyS96MjU2zKQ9_5Yyb-2JcGscVGzzsTSjJ1W-4P5SAW8c0N5fo9lt-n0zj8phekmfRjBheneI5ufl6-evie3v949vVxefr1gmtl5brwIXh1NLgPfTccas0RKBG2vowy20EpzyTDqJlGiAob33XadCi80Gckw_HvoeS79aAyzAldGEczRzyigMDyTgoVeG7_-BtXstcZxuU6kCC7HlF7RG5khFLiMOhpKmuYmB02K4w_HOF6t-cmq52Cv5Rn9Ze6-9PdYPOjLGY2SV8ZJz1FGCb7e2RRZMH87tUcvOTUyYoVwq47MQfTeeZMQ</recordid><startdate>19980201</startdate><enddate>19980201</enddate><creator>Ahmad, S</creator><creator>Selvapandiyan, a</creator><creator>Bhatnagar, R.K</creator><general>Springer</general><general>Springer Nature B.V</general><scope>FBQ</scope><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>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7QO</scope></search><sort><creationdate>19980201</creationdate><title>Increased toxicity of modified mosquitocidal binary toxins of Bacillus sphaericus expressed in Escherichia coli</title><author>Ahmad, S ; Selvapandiyan, a ; Bhatnagar, R.K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c399t-29e23a20b0edd672c2b896f60a5b60a1b2bf6c8d15c6fb1966e8dbd4496934de3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>animal health</topic><topic>animal parasites and pests</topic><topic>Animals</topic><topic>Aquatic insects</topic><topic>arthropod pests</topic><topic>Bacillus - genetics</topic><topic>Bacteria</topic><topic>Bacterial Toxins - biosynthesis</topic><topic>Bacterial Toxins - genetics</topic><topic>Bacterial Toxins - toxicity</topic><topic>Biological and medical sciences</topic><topic>Biological Sciences</topic><topic>Biotechnology</topic><topic>Crystals</topic><topic>Culex</topic><topic>E coli</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzymes</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Larva</topic><topic>Larvae</topic><topic>Methods. Procedures. Technologies</topic><topic>Microbiology</topic><topic>Modification of gene expression level</topic><topic>Plasmids</topic><topic>Proteins</topic><topic>Recombinant Proteins - biosynthesis</topic><topic>Recombinant Proteins - toxicity</topic><topic>Toxicity</topic><topic>Toxins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ahmad, S</creatorcontrib><creatorcontrib>Selvapandiyan, a</creatorcontrib><creatorcontrib>Bhatnagar, R.K</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>ABI/INFORM Collection</collection><collection>ABI/INFORM Global (PDF only)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ABI/INFORM Global (Alumni Edition)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science 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>ABI/INFORM Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Business Premium Collection</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>Business Premium Collection (Alumni)</collection><collection>Health Research Premium Collection</collection><collection>ABI/INFORM Global (Corporate)</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Business Collection (Alumni Edition)</collection><collection>ProQuest Business Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ABI/INFORM Professional Advanced</collection><collection>ProQuest Biological Science Collection</collection><collection>ABI/INFORM Global</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>One Business (ProQuest)</collection><collection>ProQuest One Business (Alumni)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Biotechnology Research Abstracts</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ahmad, S</au><au>Selvapandiyan, a</au><au>Bhatnagar, R.K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Increased toxicity of modified mosquitocidal binary toxins of Bacillus sphaericus expressed in Escherichia coli</atitle><jtitle>Applied microbiology and biotechnology</jtitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>1998-02-01</date><risdate>1998</risdate><volume>49</volume><issue>2</issue><spage>164</spage><epage>167</epage><pages>164-167</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><coden>AMBIDG</coden><abstract>The binary mosquitocidal genes of 51-kDa and 42-kDa proteins isolated from Bacillus sphaericus 1593 have been expressed at moderate levels in Escherichia coli employing the pQE expression system. The expressed proteins are readily visible in Coomassie-blue-stained protein gels. The recombinant E. coli cells expressing toxic proteins were toxic towards Culex larvae. During the assembly of crystals in B. sphaericus, the 42-kDa toxin is first cleaved at the N-terminal end by a specific B. sphaericus protease. To express the toxins in E. coli the B. sphaericus specific protease-recognition site was deleted at the N-terminal end of the 42-kDa toxin, thereby mimicking the structure of the toxin as present in the crystal. This modification resulted in a twofold increase in the toxicity of the E. coli cells expressing the modified 42-kDa toxin as a constituent of the binary toxin. Our results demonstrate the utility of this modification for heterologous expression of the binary toxin genes from B. sphaericus.</abstract><cop>Berlin</cop><pub>Springer</pub><pmid>9534256</pmid><doi>10.1007/s002530051153</doi><tpages>4</tpages></addata></record> |
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| subjects | animal health animal parasites and pests Animals Aquatic insects arthropod pests Bacillus - genetics Bacteria Bacterial Toxins - biosynthesis Bacterial Toxins - genetics Bacterial Toxins - toxicity Biological and medical sciences Biological Sciences Biotechnology Crystals Culex E coli Electrophoresis, Polyacrylamide Gel Enzymes Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Expression Genetic engineering Genetic technics Larva Larvae Methods. Procedures. Technologies Microbiology Modification of gene expression level Plasmids Proteins Recombinant Proteins - biosynthesis Recombinant Proteins - toxicity Toxicity Toxins |
| title | Increased toxicity of modified mosquitocidal binary toxins of Bacillus sphaericus expressed in Escherichia coli |
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