OL-031 In vivo investigation of Killed Leishmania Vaccine's (KLV) efficacy with Imiquimod (IMQ) as adjuvant in inhibition of visceralization of Leishmania major in Balb/c model

Abstract

Background: Escherichia coli O157:H7 is associated with hemorrhagic colitis, thrombotic thrombocytopenic purpura, and hemolyticuremic syndrome in humans. In addition to producing Shiga-like (Vero) toxin and enterohemolysin, E. coli O157:H7 has been shown to attach to the cytoplasmic membranes of intestinal epithelial cells, to efface their microvilli, and to cause actin to accumulate beneath sites of bacterial attachment. These features are shared with several other enterohemorrhagic E. coli (EHEC) serotypes and members of the enteropathogenic E. coli (EPEC) group. The eae gene, which has been shown to be necessary for attaching and effacing activity, encodes a 94to 97-kDa outer membrane protein (OMP) which is termed intimin. This gene is located in a chromosomal pathogenicity island also known as the locus of enterocyte effacement (LEE). The gold aim of this study was cloning of eae gene of E. coli O157:T in E. coli TOP10F strain. Methods: The eae gene was isolated from total genome of E. coli O157:T by PCR and eae specific PCR primers. DNA fragment of eae gene was cloned by T/A cloning technique in pGEMT easy vector (Invitrogen, San Diego, Calif.), and this construct transformed into E. coli TOP10F strain. Results: The results show that, eae was cloned in E. coli successfully. The sequencing result confirm that eae gene was cloned is correct and BLAST outcome demonstrated the sequences of eae is approved. Conclusions: Therefore it seems that the DNA constract that was produced in this study can be used for DNA vaccine against eae of E. coli O157:T in future researchs.