Expressão em Escherichia coli de Quitinase (E.C. 3.2.1.14) de Chromobacterium violaceum

Abstract

The sequencing of the Chromobacterium violaceum genome generated the necessity of characterizes the genetic function of the transcripts present in this bacterial chromosome set. In this genome were found several ORFs (Open Read Frame) corresponding to the biotechnological target genes, and among them five that codify Chitinases. This work shows the results obtained with cloning and express chitinases genes of Chromobacterium violaceum We selected four ORFs who s the DNA sequences were obtained by PCR, with specific primers. Two strategies were utilized in this work, one was cloning the complete gene sequence, and other was cloning the code region in a fusion with the Staphylococcus aureus B subunit of A protein, using the transcription and translation signals of a known expression vector. The first strategy didn t produce good results because we didn t obtained success with cloning. The second strategy it made possible the expression of the sequence corresponding to ORF CV2736, who s the enzyme was obtained in high levels and presented activity when in contact with substrate.