Engineering, Cloning and Expression of Interleukin 2–Com1 Chimera with Aim of Recombinant Subunit Vaccine Production Against Coxiella burnetii

Abstract

Query fever is an important disease caused by Coxiella burnetii, therefore vaccination against this disease is so crucial. Com1 is one the most important immunogenic proteins of C. burnetii which can be appropriate candidate to design a subunit vaccine. It seems, fusion of this protein with interleukin 2 as a molecular adjuvant can promote its efficacy. To do current study, first, Com1 and interleukin 2 genes were individually amplified by PCR, then these PCR products were applied to fuse interleukin 2 with Com1 using splice overlap extension PCR. The product of splice overlap extension PCR was cloned in pTZ57R/T vector by T/A cloning strategy, after digestion, interleukin 2–Com1 gene was sub-cloned in pET-22b(+) by T4 DNA ligase enzyme. Ligation product was applied to express in BL21 (DE3) strain of Escherichia coli. Expressed interleukin 2–Com1 protein was purified by Nik–NTA affinity column and then confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. The results of electrophoresis on agarose 1% gel revealed that, PCR of target genes and splice overlap extension PCR were successfully performed. The results of electrophoresis on 12% sodium dodecyl sulfate polyacrylamide gel confirmed expression and purification of interleukin 2–Com1 protein. Finally, the results of western blotting shown, purified protein with a molecular size of 43.5 kDa belonged IL2–Com1 chimera. The results of present study shown, BL21 (DE3) is an appropriate host to express interleukin 2–Com1 protein. It seems, this chimera can be introduced as a potent candidate to fight with Query fever.