Cloning and expression of a novel cry gene that is potentially active against nematodes

Autor: Flvio Figueiredo Barbosa Bruno, Victor Franco Lemos Manoel, Lucrecio da Costa Alisson, Maia dos Santos Jaime, Fernandes Gonalves Janana, Boter Bergamasco Vivian, Luiz de Laia Marcelo
Rok vydání: 2014
Předmět:
Zdroj: African Journal of Microbiology Research. 8:1017-1025
ISSN: 1996-0808
DOI: 10.5897/ajmr2013.6282
Popis: Biological control may reduce nematode populations to acceptable levels using Bacillus thuringiensis. Thus, the search for new cry proteins that are active on this type of target is highly relevant. To identify novel cry genes, B. thuringiensis isolates were analysed using molecular biology. The isolates with DNA sequences resembling genes that are effective against nematodes were fully sequenced through primer walking. The characterised novel cry gene was cloned in an expression vector, and the target protein was expressed at in an appropriate E. coli strain. The highest levels of protein expression were expressed at 18°C. To discern the nematotoxicity of this protein, bioassays were conducted using free living nematodes as targets. The bioassays revealed that the assessed free living nematodes were susceptible to the analysed cry protein. These results indicate that nematodes can be targets of cry protein produced by isolates of this bacterium. Key words: Biological control, primer walking, cry proteins, bioassays.
Databáze: OpenAIRE