Molecular cloning, characterization, and homologous expression of an endochitinase gene from Bacillus thuringiensis serovar morrisoni

Authors : Sezer OKAY, Gülay ÖZCENGİZ

Pages : 1-7

View : 19 | Download : 7

Publication Date : 2011-12-01

Article Type : Research Paper

Abstract :The endochitinase gene insert ignore into journalissuearticles values(chi3023); of Bacillus thuringiensis insert ignore into journalissuearticles values(Bt); serovar morrisoni strain 3023 was amplified via polymerase chain reaction insert ignore into journalissuearticles values(PCR); and cloned in Escherichia coli. The ORF of chi3023 insert ignore into journalissuearticles values(GenBank Accession Number: DQ993175); consists of 2031 nucleotides encoding a 676-residue protein with a calculated molecular mass of 74.5 kDa and a pI value of 6.0. The amino acid sequence of Chi3023 was compared with previously sequenced Bt chitinases and the phylogenetic relationships among them were determined. The deduced N-terminal 34 amino acids of the premature Chi3023 exhibited a typical signal peptide. The E. coli-Bt shuttle vector pHT315 was used for homologous expression of chi3023. Introduction of recombinant pHT315BTC, carrying chi3023 into Bt serovar morrisoni 3023, resulted in a 23-fold increase in endochitinase activity insert ignore into journalissuearticles values(0.185 U/mg versus 4.256 U/mg);.
Keywords : Key words Bacillus thuringiensis, characterization analysis, endochitinase, gene cloning