Cloning, expression and purification of antigenic fragments of the Ureaplasma urealyticum UreD protein and its value in serology

Faleh Darvishi; Ali Ganji; Behzad Khansarinejad; Shabnam Sadoogh Abbasian; Hamid Abtahi

doi:10.18502/ijm.v14i6.11255

Faleh Darvishi Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran
Ali Ganji Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran
Behzad Khansarinejad Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran
Shabnam Sadoogh Abbasian Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran
Hamid Abtahi Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran

DOI: https://doi.org/10.18502/ijm.v14i6.11255

Keywords: Ureaplasma urealyticum; Recombinant proteins; Blotting; Western; Epitopes; Cloning; Molecular; Vaccines

Abstract

Background and Objectives: The detection of Ureaplasma urealyticum is usually done through culture. With the change of the smallest effective factor inculture, we face the lack of growth of these bacteria, which is one of the important reasons to find a suitable alternative for the diagnosis of this bacterium. UreDis a protected gene in this bacterium. The aim of this was to evaluate the ability of antigenic regions of UreD protein to bind to patients' serum antibodies.

Materials and Methods: Antigenic regions of UreD protein were predicted using IEDB software with five different meth- ods: Emini Surface Accessibility Prediction, Kolaskar and Tongaonkar Antigenicity, Chou and Fasman beta turn prediction, Karplus and Schulz flexibility scale, Ellipro –Epitope predictionbased upon structural protrusion. Antigenic regions of UreD gene was clonned, expressed and purified. The antigenicity of this recombinant protein against theantibodies in the serum of people infected with U. urealyticum infections was checked in western blotting.

Results: The results showed that the antigenic regions of the UreD protein was producted and its antigenicity was demon- strated in western blotting.Moreover, all sera from patients infected with U. urealyticum reacted to the recombinant antigen.

Conclusion: Specimens from people infected with U.urealyticum infection was positive in Western blotting suggesting that UreD protein has antigenic properties. Therefore, it can be used as a suitable candidatefor the design of diagnostic kits and U. urealyticum vaccine.