Page 67 - WINGS OF AGRI-INNOVATIONS

26. Biopesticidal Formulation With Improved Shelf Life And The Process Of Preparation 27. Synthetic Gene Encoding A Chimeric Delta-Endotoxin Of Bacillus Thuringiensis

Patent Number 234480 Patent Number 237912
Date Of Certificate Issue 15/06/2009 Date Of Certificate Issue 14/01/2010
Post Grant Journal Date 26/06/2009 Post Grant Journal Date 22/01/2010
Publication Number 33/2007 Publication Number 14/2008
Publication Date 17/08/2007 Publication Date 04/04/2008
Publication Type INA Publication Type INA
Application Number 405/DEL/2006 Application Number 2048/DEL/2006
Application Filing Date 14/02/2006 Application Filing Date 18/09/2006
Field Of Invention MICRO BIOLOGY Field Of Invention BIOTECHNOLOGY

Classification (IPC) A01N 63/00 Classification (IPC) C12N15/09

Inventor DR. PREM DUREJA, DR. BALRAJ SINGH PARMAR Inventor POLUMETLA ANANDA KUMAR
Abstract:
Abstract:
A process for preparation of ready to use solid biopesticidal formulations with improved shelf life based on The present invention provides a means for plant protection against damage by a broad spectrum of insect
pests. The invention comprises a chemically synthesized gene encoding a unique chimeric insecticidal
the biocontrol agent(s) such as the antagonist mycoparasite(s) ex. Trichoderma species and the biopesticidal protein (δ-endotoxin) of Bacillus thuringiensis. The chimeric δ-endotoxin comprises of Domain I of CrylAal,
product obtained thereof are reported. The dried and powdered solid hyphae of the agent are blended Domain II of CrylAbl and part of Domain III of CrylAcl and a part of Domain III of CrylFal. The unique chimeric
directly with an inorganic or organic inert carrier with or without the addition of other formulant(s) to protein is 624 amino acids in length. A synthetic gene was constructed based on DNA sequences of the native
obtain a ready to use product complying the application requirements for use in pest control. It passed Bt structural genes (crylAal, crylAbl, crylAcl and crylFal). The sequences were modified in order to contain
the accelerated storage test (54 ± 1°C, FAO 1999) prescribed for assessing the shelf life of two years of the codons preferred by plant genes, to attain an A+T content in nucleotide base composition found in plants,
chemical pesticides.
and to eliminate sequences that cause destabilization. polyadenylation, degradation and termination of
RNA and to avoid sequences that constitute RNA splice sites. In the synthetic gene codons used to specify
a given amino acid were selected with regard to the distribution frequency of codon usage employed by
plant genes to specify that amino acid. In addition, consideration was given to the percentage G+C content
of the degenerate third base. The Bt gene designed for enhanced expression in plants was synthesized in
fragments and assembled within a DNA plasmid vector. The synthetic Bt gene was then introduced into a
tobacco plant and expressed. The chimeric insecticidal protein produced upon expression of the synthetic
Bt gene in tobacco plants exhibited toxicity to Cotton bollworm and Tobacco caterpillar.

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