Construction Trichinella Spiralis45Ku ES Rcombinant Eukaryotic Plasmids And Detection Vaccination Effects

Trichinella spiralis is one of the most widespread parasites which can infect human and animals.Trichinellosis caused by T.spiralis is a serious parasitic zoonosis with worldwide distribution and had seriously influenced human production and life. In order to prevent and control trichinosis, scholars had done a lot of research work on trichinosis immune preventive.with the development of modern molecular biology technology,the DNA vaccines was produced by gene engineering technology can solove the problems of the first generation vaccine. This experiment had used muscle larvae45ku ES antigen gene construct the nucleic acid vaccine and test the immune effect.According to45ku ES antigens of Trichinella spiralis muscle larvae gene sequence form GenBank。We design a pair of primer.Introduced two restriction sites. Cloning target gene by PCR.Digestion the plasmid which is extract from positive bacteria. Construction of recombinant plasmid pVAX1-Ts45.The recombinant plasmid was transformed into COS-7cells. We can found the plasmid can mixis with COS-7cells and expression by the fluorescence microscopy.Three groups of BALB/c mice were respectively immunized by restructuring plasmid pVAX1-Ts45、PBS、pVAX1for three times, every time interval a week.Then the mice were challenged by gavage administration with trichinella spiralis muscle larvae. Thrity days later, the protective effects of the recombinant was determined by measuring body weight change, worm reduction rates, IgG antibody titers in serum, the level of cytokine(IL-4,IFN-y) in PBMC and the frequency of main lymphocytes(CD4+CD25+Foxp3+T cells) in MLN and spleen. The results showed that immunization with DNA vaccine resulted in significantly improved body weight gain, worm reduction rates, histopathology compared to other groups. The Worm reduction of DNA Vaccine group is29.8%. Furthermore, recombinant DNA vaccine leads to a significant increase in serum antibody titers, the level of cytokine and the activity of main lymphocytes have significant increase.