Construction Of TgDLC8a-GST Polymer Protein Complex Expression Vector And Its Application On Chicken Coccidiosis Vaccine

Homologous dimer is a common form of biological protein.The two homologous dimers can form polymeric complexes.It can be widely used in many fields,and one of the most important applications is that it can be used as an antigen expression platform to insert antigen epitope to increase the load of antigens.This research use the power of toxoplasma protein light chain 8 a(TgDLC8a)and glutathione S transferase(GST)this two homologous dimers expression to form polymer compound,as a vaccine expression platform,which insert the chicken Amy's ball insect various antigen fragments.PGEX-6P-1 was used as the carrier to clone TgDLC8 a gene,and the GST protein with PGEX-6P-1 was used to complete the fusion with TgDLC8 a.In order to show the antigen,selection of GST and TgDLC8 a genes between BamHI and TgDLC8 a protein spatial structure affected parts(37-42 bp)two positions as insertional locus of antigen,build more express carrier TgDLC8a-GST.The key structural domain of the emmeric antigen EtMIC2?EtMIC1H was cloned into the mu TgDLC8a-GST vector,and the expression platform was evaluated by the multivalent antigen complex induced expression.TgDLC8a-GST transmutation of BL21 induced expression products via Native-PAGE and Western-blot analysis showed that the fused gst-tgdlc8 a homologous dimer could form a polymeric complex.After antigen gene cloning and recombinant expression vector can be successfully protein expression and form dimers and polymer,suggests TgDLC8 a inserted enzyme sites does not affect the homologous dimers formation,the platform can be used for the vaccine.Studies have shown that the homologous dimers formed by TgDLC8 a and GST can form polycomplexes.This study successfully constructed the recombinant polymeric expression platform(TgDLC8a-GST),which can effectively display the antigens in the form of polyvalent states and provide a good platform for the research and development of genetic engineering vaccines.The immune protection effect of this recombinant vaccine strain was investigated by using chicken coccidiosis test.120 a day old chicks were randomly divided into an average of six group,group I and II for the PBS control group,group III,IV,V,VI animal skin more immune TgDLC8a-GST protein antigen vaccine,TgDLC8a-GST-MIC2-MIC1 H protein antigen vaccine,EtMIC2 protein antigen vaccine,EtMIC1 H protein antigen vaccine.At age21,16,000 E.tenella spore oocysts were administered to II,III,IV,V and VI groups.At the age of 31,the results were analyzed.Animal test results showed that IV,V,VI group protein antigen vaccine could provide a certain protective effect to the infection of coccidioides,and the expression of vaccine strains in the IV group was more effective than that of V and VI groups.