Engineered Cry1Ac-Cry9Aa hybrid Bacillus thuringiensis delta-endotoxin with improved insecticidal activity against Helicoverpa armigera.

Recombinant Bt construct was prepared by exchange of pore forming domain I with cry1Ac to cry9Aa gene by overlap extension PCR (OE-PCR) technique. Construction of cry1Ac-cry9Aa was accomplished by six base pair homology at 3' ends of PCR products of domain I of cry1Ac and domain II and III of cry9Aa. The recombinant toxin was also modified by deletion of N-terminal alpha helix-1 of recombinant toxin.

Molecular Characterization of Novel Serovars of Bacillus thuringiensis Isolates from India.

Novel Bacillus thuringiensis isolates GS4, GN24 and UP1 were isolated and characterized by determination of serotyping, insecticidal protein by SDS-PAGE, plasmid composition, cry gene content and insect toxicity. Serologically two isolates GS4 and UP1 were allocated to the H3abce which is a new serovar while isolate GN24 was of H3ab type. Isolate GS4 produced flat crystal inclusions while UP1 produced cuboidal crystals.

A new enrichment method for isolation of Bacillus thuringiensis from diverse sample types.

New or more efficient methodologies having different principles are needed, as one method could not be suitable for isolation of organisms from samples of diverse types and from various environments. In present investigation, growth kinetics study revealed a higher germination rate, a higher growth rate, and maximum sporulation of Bacillus thuringiensis (Bt) compared to other Bacillus species. Considering these facts, a simple and efficient enrichment method was devised which allowed propagation of spores and vegetative cells of Bt and thereby increased Bt cell population proportionately.

Distribution and diversity analysis of Bacillus thuringiensis cry genes in different soil types and geographical regions of India.

Molecular characterization of 117 Bacillus thuringiensis (Bt) isolates from various geographical locations was previously done by PCR amplification of cry genes. In present investigation, diversity of cry genes from different soil types and climatic environments was studied using rarefaction method. Presence of cry1, cry2, cry3, 7, 8, cry4, cry5, 12, 14, 21, cry11, cry13 and cyt1 genes from Bt strains isolated from various regions of India was determined by PCR amplification.

RFLP analysis of cry1 and cry2 genes of Bacillus thuringiensis isolates from India.

The PCR-RFLP method has been useful for detection of known genes and identification of novel genes. In the present study, degenerate primers were designed from five groups of cry1 genes for PCR-RFLP analysis. Bacillus thuringiensis (Bt) isolates from different regions were evaluated for PCR amplification of various cry1 genes using newly designed primers and cry2 genes using reported primers.

Molecular characterization of Bacillus thuringiensis isolated from diverse habitats of India.

Bacillus thuringiensis (Bt) strains were isolated from 94 samples from different geographical regions. Novel types of crystalline inclusion bodies were observed from some of the isolates. Crystalline inclusions of bipyramidal, spherical and cuboidal morphology were found produced by most of the isolates.