Molecular cloning and expression of Bacillus anthracis Lethal Factor domain 1 gene in Escherichia coli

Anthrax is a common disease in human and livestock caused by Bacillus anthracis. Bacillus anthracis has two strong immunogen proteins: Protective antigen (PA) and lethal factor domain I (LFD1) that has been always considered as a candidate vaccine against Bacillus anthracis. The aim of this study was to express the lethal factor domain I in Escherichia coli. In this laboratory experimental study, the gene of LFD1 was detected and amplified from pXO1 plasmid by PCR. The gene was cloned with Bam H I and Xho I restriction site in cloning vector (pGEM-T easy), after isolation was sub cloned to expression vector pET28a(+). This vector was transformed to E. coli-BL21 (DE3) to express LFD1 gene. The expression of LFD1 gene was induced by IPTG, and LFD1 protein was produced. The cloned LFD1 gene in pET28a(+) vector was confirmed by sequencing, PCR and enzymatic analysis. The expressed recombinant protein was confirmed by SDS-PAGE and Western blotting. According to immunogenicity of LFD1 protein, the produced recombinant protein can be used separately or in combination by adjuvants and delivers to design a vaccine against anthrax.