Page 69 - WINGS OF AGRI-INNOVATIONS

28. Additives For Improved Photostability Of Azadirachtin-At 29. Synthetic Gene Encoding Cry1fa1 Delta - Endotoxin Of Bacillus Thuringiensis

Patent Number 227536 Patent Number 242768
Date Of Certificate Issue 26/03/2010 Date Of Certificate Issue 09/09/2010

Post Grant Journal Date 27/03/2009 Post Grant Journal Date 17/09/2010
Publication Number 25/2008 Publication Number 14/2008
Publication Date 20/06/2008 Publication Date 04/04/2008
Publication Type INA Publication Type INA
Application Number 460/DEL/2001 Application Number 2049/DEL/2006
Application Filing Date 04/04/2001 Application Filing Date 18/09/2006
Field Of Invention AGROCHEMICALS Field Of Invention BIOTECHNOLOGY

Classification (IPC) A01N 65/00 Classification (IPC) C12N15/09
Inventor SAPNA JOHNSON, PREM DUREJA, Inventor POLUMETLA ANANDA KUMAR
SWARAN DHINGRA Abstract:
Abstract: The present invention provides a means for plant protection against aarrrage to crops, especially to
The effectiveness. persistence and scope of use of many pesticides are limited or restricted by their stability, tobacco and brinjal by insect pests, especially Spodopteralitura and Leucinodesorbonalis. The invention
particularly to sunlight. The present invention relates to a process for preparation of photostable technical comprises a chemically synthesized gene encoding an insecticidal protein namely Cry lFal (endotoxin). The
azadirachtin-A concentrate (a neem based pesticide), for use in the pest control of Spodoptera litura in the CrylFal 5-endotoxin protein is 616 amino acids in length. A synthetic gene was constructed based on DNA
field by use of an effective amount of toxicologically safe additives to photolabile technical azadirachtin-A sequences of the native Bt structural genes (crylFal). The sequences were modified in order to contain
concentrate. codons preferred by plant genes, to attain an A+T content in nucleotide base composition found in plants,
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
tobacco and brinjal plants and expressed. The CrylFal insecticidal protein produced upon expression of the
synthetic Bt gene in tobacco and brinjal plants exhibited toxicity to Tobacco caterpillar and Brinjal Shoot
and Fruit borer.

60 61