Preparation Of Monoclonal Antibodies Against ROP4 And ROP5 Protein Of Toxoplasma Gondii And Its Character Identification

Toxoplasma gondii(T.gondii),an opportunistic protozoan parasite,infects almost all warm-blood animals and causes toxopiasmosis of the host.The WHO and FAO have recently established it as a foodborne parasite infection causing global concern.It is a serious threat to human health especially pregnant women and immunocompromised individuals.T.gondii can lead to abortion,monster and stillbirth during gestation period.Moreover it is one of the factors causing death for immunocompromised patients.As an opportunistic infection factor,T.gondii can result in serious organ lesion on the patients such as HIV,organ transplant,and cancer.In addition,the diseases of livestock and poultry caused by toxoplasma contributes to tremendous economic losses.The consequences of toxoplasmosis infection which persecuted us to pay much more attention to study the diagnosis of toxoplasmosis.The traditional diagnosis methods have many disadvantages respectively.The monoclonal antibody has strong specificity and it is has been used in the rearch of T.gondii.In this study,we focused on T.gondii ME49 strain,and the objectives of which were to clone the rhoptry protein 4(ROP4)gene.A recombinant prokaryotic expression vector pET32a-ROP4 was successfully constructed,then was transformed into E.coli Rosetta(DE3)and possessed the specific reactionogenicity via Western blot.Utilized the optimized expression condition to express objective proteion and then purified the proteion.Immunized mices useing purified proteion to make monclonal antibody.Two strains of monoclonal antibodies anti-recombinant truncated ROP4 named G-2 and 3B-7 were prepared,which all reacted with polypide or native ROP4 by ELISA IFA and Western-blot.Also two strains of monoclonal antibodies anti-recombinant truncated ROP5 named 2B-7 and 2B-2 were prepared used the same programs as ROP4.