Novel insecticidal chitinase protein its encoding nucleotide and application thereof

Abstract

A novel insecticidal chitinase protein from fern Tectaria sp., a process for preparation of the insecticidal protein and nucleic acid sequence encoding for said insecticidal protein and its application for insect control purposes.

1 . A novel insecticidal chitinase protein, containing chitin binding module but lacking catalytic module wherein the nucleotide sequence encoding the insecticidal protein is represented by any one of the SEQ ID No.1 or SEQ ID No. 2 or SEQ ID No. 3 and amino acid sequence of the insecticidal protein is represented by any one of the sequences SEQ ID No. 4 or SEQ ID No. 5. 
     
     
         2 . The insecticidal protein as claimed in  claim 1 , wherein said insecticidal protein is of 216 amino acid residues long pro-protein and 192 amino acid residues long mature protein with respective molecular weight 23.684 and 21.270 kDa. 
     
     
         3 . The insecticidal protein as claimed in  claim 1 , wherein said protein comprises of chitin binding module (CtBM), and shows exo- and endo-chitinase activity. 
     
     
         4 . The insecticidal protein as claimed in  claim 1 , wherein the said protein consists of chitin binding module (CtBM), and shows exo- and endo-chitinase activity. 
     
     
         5 . The insecticidal protein as claimed in  claim 1 , wherein the said protein is useful for the control of insects from order homoptera, heteroptera, diptera, coleoptera and Lepidoptera, particularly toxic to white fly ( Bemisia tabaci ). 
     
     
         6 . A process for preparation of the insecticidal chitinase protein as claimed in  claim 1 , comprising the steps of:
 (i) isolating chitinase protein from fern  Tectaria  sp. in a manner such as herein described,   (ii) cloning c-DNA from purified protein, using N-terminal sequencing data of the purified protein by designing degenerate primers,   (iii) identifying ORF sequence encoding mature polypeptide of insecticidal chitinase from cloned cDNA sequence,   (iv) cloning the DNA encoding the insecticidal protein in  E. coli  expression vector in fusion with SUMO peptide to get expression of recombinant protein followed by purification of recombinant protein by conventional manner.   
     
     
         7 . The process as claimed in  claim 1 , wherein the insecticidal protein is being produced by expressing its encoding nucleotide in homologous or heterologous system using recombinant DNA technology. 
     
     
         8 . The process as claimed in  claim 1 , wherein the nucleotide sequence of insecticidal protein obtained by the process is useful to produce transgenic crop plants, as herein described, which express the insecticidal protein causing toxicity to insect and exhibiting protection against insect pest. 
     
     
         9 . An expression vector as claimed in  claim 6 , comprising of the nucleotide sequence of the insecticidal protein and used to get expression of recombinant protein. 
     
     
         10 . Use of the insecticidal protein as claimed in  claim 1  in control of insects.