The Preliminary Research Of M.suis MSG1 Recombiant Protein And PVAX1-MSG1 Nucleic Acid Vaccine Combined Immunization

Anemia,jaundice and other symptoms were caused by Mycoplasma suis(M.suis)which spread among pigs.And animals immunity were decreased,growth was slowed and weight was losed,which can lead to animal deaths.Drugs were not able to complete treatmented and vaccines were not preventived on the market currently.Preparation of MSG1 protein and nucleic acid vaccine,combined immunization and single immunizaty for research,and starts from the following 3 aspects:1.MSG1 gene was obtained by PCR,homology alignment was blasted in NCBI after sequencing,and bioinformatics software was used to analyze the MSG1.Results showed exactly the same homology of MSG1 was a complete open reading frame,the sequence was 1011 bp,encoded 336 amino acids,no signal peptide and transmembrane domain,and optimization software was used to optimize codons.2.Target gene was connected with prokaryotic expression vector pET-28(b+)and pVAX1 eukaryotic expression vector,the conditions of protein expression and purification of nucleic acid vaccines were optimized.Results showed that the protein could be obtained in supernatant,and the purified protein was validated,the preparation of nucleic acid vaccine was purity.3.After mice was immunized,SOD and MDA were determinated to evaluate the oxidative damage of body.ALT,AST,CREA,UREA were measured by biochemical parameters to evaluate the damage of liver and kidney,Tbil,Dbil,Glu were analysed for the determination of MSG1 associated with jaundice,anemia.IL2 were determinate to evaluate the level of cell-mediated immunity,ELISA method were established to evaluate the level of humoral immunity.Results showed that they does not have oxidative damage in the body in different immune,and they does not affect the hepatic and renal function of MSG1 has no direct contact with anemia,jaundice.The humoral immunity level could be increased by the protein of MSG1,but the level of cell-mediated immunity could not be increased.The level of cell-mediated immunity could be increased by nucleic acid vaccine,but the level of humoral immunity was increased a little,and the level of cell-mediated immunity and humoral immunity were increased by combined immunization.