Abstract
Recombinant Gluconacetobacter diazotrophicus containing Cry1Ac gene from Bacillus thuringiensis var. kurstaki borne on pKT230, shuttle vector, was generated. PCR amplification of Cry1Ac gene present in recombinant G. diazotrophicus yielded a 278-bp DNA product. The nitrogenase assay has revealed that the recombinant G. diazotrophicus in sugarcane stem produced similar levels of nitrogenase compared to wild-type G. diazotrophicus. The presence of 130-kDa protein in apoplastic fluid from sugarcane stem harvested from pots inoculated with recombinant G. diazotrophicus shows that the translocated G. diazotrophicus produces 130-kDa protein which is recognized by the hyperimmune antiserum raised against 130-kDa protein. The first instar Eldana saccharina neonate larvae that fed on artificial medium containing recombinant G. diazotrophicus died within 72 h after incubation.
Copyright © 2011 S. Karger AG, Basel.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Bacillus thuringiensis / genetics*
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Bacillus thuringiensis / metabolism
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Bacillus thuringiensis Toxins
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Bacterial Proteins* / genetics
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Bacterial Proteins* / metabolism
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Electrophoresis, Polyacrylamide Gel
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Endotoxins* / genetics
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Endotoxins* / metabolism
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Gluconacetobacter / genetics*
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Gluconacetobacter / metabolism
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Hemolysin Proteins* / genetics
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Hemolysin Proteins* / metabolism
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Immunoblotting
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India
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Larva / microbiology
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Lepidoptera / growth & development
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Lepidoptera / microbiology
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Microscopy, Electron, Scanning
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Nitrogen Fixation*
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Nitrogenase / genetics
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Nitrogenase / metabolism
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Pest Control, Biological*
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Polymerase Chain Reaction
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Recombination, Genetic*
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Saccharum / metabolism
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Saccharum / microbiology
Substances
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Bacillus thuringiensis Toxins
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Bacterial Proteins
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Endotoxins
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Hemolysin Proteins
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insecticidal crystal protein, Bacillus Thuringiensis
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Nitrogenase