EXPRESSION AND PURIFICATION OF AN IRON SCAVENGER RECEPTOR OF PROTEUS MIRABILIS AS A NEW TARGET AGAINST URINARY TRACT INFECTIONS

Background and Aim:Proteus mirabilis strains are among the most common causes of urinary tract infections. Iron adsorption receptors of P. mirabilis strains have the properties of an ideal vaccine candidate against UTIs. In the present study, amplification, expression and purification of iron adsorption receptor PMI0842 of P. mirabilis was performed to evaluate its efficacy in other studies.Methods:The amplification of PMI0842 gene in P. mirabilis HI4320 isolate was performed by using the Polymerase Chain Reaction (PCR). Then, the amplified gene was cloned and expressed in pET28a-BL21 (DE3) expression system. Purification of the PMI0842 expressed protein was performed by nickel column under the denaturation conditions. Finally, the purification of recombinant protein PMI0842 was evaluated by SDS-PAGE and Western blot.Results:The PMI0842 gene was amplified with a size about 2000 bp in PCR. Confirmation the cloning of PMI0842 gene in pET28a vector was done using colony PCR, double enzyme digestion, and sequencing. Then, expression of PMI0842 was performed in BL21 (DE3) that SDS-PAGE and western blot showed the high purity of the eluted protein on column. The approximate size of the purified protein was about 70 KD.Conclusion:There is currently no effective vaccine available against UTIs; therefore, development of an ideal against these infections is urgently required. Iron scavenger receptors could be among the ideal candidates against P. mirabilis that efficacy some of them were evaluated against UTIs. In this study, PMI0842 was purified for the first time and evaluation of its efficacy is under study.

Keywords:

Urinary tract infection , P. mirabilis , PMI0842 , Vaccine candidate