Document Detail


High-level cryIVD and cytA gene expression in Bacillus thuringiensis does not require the 20-kilodalton protein, and the coexpressed gene products are synergistic in their toxicity to mosquitoes.
MedLine Citation:
PMID:  8481007     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Interactions among the 20-kDa protein gene and the cytA and cryIVD genes located in a 9.4-kb HindIII fragment were studied. A series of plasmids containing a combination of these different genes was constructed by using the Escherichia coli/Bacillus thuringiensis shuttle vector pHT3101. The plasmids were then used to transform an acrystalliferous strain, cryB, derived from B. thuringiensis subsp. kurstaki. The results from sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analyses suggest that although the 20-kDa protein is required for the efficient CytA protein production in E. coli, it is not required in B. thuringiensis. With or without the truncated 20-kDa protein gene, the CtyA and/or CryIVD proteins are produced and form parasporal inclusions in B. thuringiensis cells. However, more-efficient expression is obtained when a second protein, probably acting as a chaperonin, is present. In addition, the time course studies show that the CytA and CryIVD proteins are coordinately produced. Both the crude B. thuringiensis culture and purified inclusions from each recombinant B. thuringiensis strain are toxic to Culex quinquefasciatus larvae. The parasporal inclusions formed in B. thuringiensis cells are mosquitocidal, with CytA synergizing CryIVD toxicity.
Authors:
C Chang; Y M Yu; S M Dai; S K Law; S S Gill
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  59     ISSN:  0099-2240     ISO Abbreviation:  Appl. Environ. Microbiol.     Publication Date:  1993 Mar 
Date Detail:
Created Date:  1993-05-27     Completed Date:  1993-05-27     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  815-21     Citation Subset:  IM    
Affiliation:
Department of Entomology, University of California, Riverside 92521.
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MeSH Terms
Descriptor/Qualifier:
Animals
Bacillus thuringiensis / genetics*
Bacterial Proteins / biosynthesis,  genetics*,  metabolism*
Bacterial Toxins / biosynthesis,  genetics*
Culex
Deoxyribonuclease HindIII
Drug Synergism
Electrophoresis, Polyacrylamide Gel
Endotoxins*
Gene Expression Regulation, Bacterial
Genes, Bacterial
Hemolysin Proteins
Immunoblotting
Pest Control, Biological
Plasmids
Grant Support
ID/Acronym/Agency:
ESO3298/ES/NIEHS NIH HHS
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/Bacterial Toxins; 0/Endotoxins; 0/Hemolysin Proteins; 0/insecticidal crystal protein, Bacillus Thuringiensis; EC 3.1.21.-/Deoxyribonuclease HindIII
Comments/Corrections

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