ABSTRACT
Chitinases genes from Metarhizium anisopliae which is an important entomopathogenic fungus were considered one of the key factors to invade their hosts. One Metarhizium anisopliae HN1 strain was isolated and screened. A chitinase gene was amplified by RT-PCR from Metarhizium anisopliae HN1, The whole length of this gene was 1275bp,and the nucleotide sequence of the gene was 96% similarity to that of the M. anisopliae E6 accessed in GenBank ( AF02749). The gene has been registered in GenBank and its accession number is DQ011865. The gene was subcloned into prokaryon expression vector pET-22b( + ), transformed this recombinant expression plasmid into E. coli strain BL 21 and effective expressed. The SDS-PAGE analysis indicated that the recombinant protein was 42kDa which is same to the reported article. The expression level of recombinant protein was about 63. 3% of whole expressed proteins , And when recombinant E. coli were crushed by freeze and supersonic wave , the activity assay indicates that the chitinase expressed in bacteria possesses biological activity.
ABSTRACT
The strains Dz01 and Ma4 were isolated from cadavers of Brontispa longissima(Gestro),and were confirmed to be pathogenicity to Brontispa longissima(Gestro).After microscopical observation of the morphological characters of mycelium,phialide and conidia from two isolates,they were found to be identical to Metarhizium anisopliae var anisopliae,so they were identificated as M.anisopliae var anisopliae.The Maximum Parsimony tree constructed based on the sequences of ITS1-5.8S-ITS2 regions in ribosomal DNA from two isolates and 31 other isolates which represent different species or varity species of genus Metarhizium obtained from GenBank database showed that two isolates clustered together in the clade which was composed of the isolates classified as Metarhizium anisopliae var anisopliae.This provided the molecular data for the result of morphological identification of Dz01 and Ma4 isolates.