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Cloning and expression of Mannheimia haemolytica PlpE gene in Escherichia coli and its immunogenicity assessment | ||
| Archives of Razi Institute | ||
| مقالات آماده انتشار، پذیرفته شده ، انتشار آنلاین از تاریخ 26 اسفند 1397 | ||
| نوع مقاله: Original Articles | ||
| شناسه دیجیتال (DOI): 10.22092/ari.2018.116479.1169 | ||
| نویسندگان | ||
Azadeh Yektaseresht   ؛ Fatemeh Sabet Sarvestani؛ Maryam Dordani؛ Arsalan Hosseini
| ||
| Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran | ||
| چکیده | ||
| Mannheimia haemolytica is responsible for considerable economic losses to cattle, sheep and goats industries in many parts of the world. M. haemolytica is one of the causative agents of shipping fever of cattle. Current vaccines against M. haemolytica are moderately efficacious. They do not provide complete protection against the disease. Production of economic vaccine for protecting farm animals against M. haemolytica has the attention of many scientists. The outer membrane proteins (OMPs) play a major role in pathogenesis and immunogenicity of M. haemolytica. Research with M. haemolytica outer membrane proteins has shown that antibodies to one particular outer membrane protein, PlpE, may be important in immune protection. Recent studies have focused on vaccines containing antigenic proteins to increase the protection range. In the current study, the gene for M. haemolytica outer membrane protein, PlpE, was cloned into the expression vector pET26-b and was then expressed in Escherichia coli BL21. The expression of protein was carried out by induction of cultured E. coli Bl21 cells with 1mM isopropyl-β-D-thiogalactopyranoside (IPTG). The recombinant PlpE was purified using Ni-NTA agarose resin and was subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The identity of the expressed protein was analyzed by western blotting. It was revealed that rPlpE was expressed and produced properly. The purified rPlpE was used as an antigen for antibody production in goat to assess the immunogenicity of recombinant protein. The observations suggested that the produced protein can be used as a candidate antigen for developing diagnostic tests and as a potential vaccine candidate. | ||
| کلیدواژه‌ها | ||
| Mannheimia hemlolytica؛ . PlpE؛ . Cloning؛ . Expression؛ . Immunogenicity | ||
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آمار تعداد مشاهده مقاله: 27 |
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