Construction and expression of aspartic protease from Onchocerca volvulus* as ompA fusion protein in a mutant strain of Salmonella typhimurium

• Main Authors: Abbas Jolodar, David J. Miller
• Format: Article
• Language: English
• Published: Sociedade Brasileira de Genética 2002-01-01
• Series: Genetics and Molecular Biology
• Subjects: filarial parasite; nematode; Salmonella typhimurium
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572002000100018

Two constructions in pHS164 vector were designed to permit expression of OV7A and OV4A inserts encoding the N-terminal and C-terminal portion of an aspartic protease from Onchocerca volvulus, respectively. A novel 39 kD protein ompA-OV7A fusion protein was stably expressed as ompA fusion in a modified strain of Salmonella typhimurium strain SL5000 and E.coli strain JM109. Expression of the fusion protein in bacterial strains harboring the constructs were evaluated by western blotting. E.coli and Salmonella lysates were fractionated by 10% SDS-PAGE gel and then immobilized to nitrocellulose membrane by electroblotting. Primary polyclonal antibody generated in rats against the GST-OV7A fusion protein was used in the Western blots. It remains to be seen whether the fusion protein expressed in vivo will promote effective immune response.