Preparation of bovine mastitis Streptococcus agalactiae PI-2a pili islands AP1-AP2-BP subunit antibody
WANG Xue-yin1,3,4,BU Ri-e1,3,4,CHEN Jin-long5,WANG Jin-liang2, WU Jin-hua1,3,4,XI LIN Gao-wa1,3,4
1. Inner Mongolia Autonomous Region Engineering Technology Research Center of Prevention and Control the Pathogenic Bacteria in Milk, Tongliao 028043,China; 2. Shandong Binzhou Animal Science and Veterinary Medicine Academy,Binzhou 256600, China; 3. College of Life Science, Inner Mongolia University for Nationalities, Tongliao 028043,China; 4. Research Institute for Pathogenic in Milk of Inner Mongolia University for Nationalities, Tongliao 028043, China; 5.Shandong Lvdu Bio-sciences and Technology Co.,LTD, Binzhou 256600, China
Abstract:In this experiment, the subunit recombinant antigen AP1-AP2-BP of the pili island PI-2a of bovine mastitis Streptococcus agalactiae was constructed and its multi-subunit antibody was prepared, which provided the experimental basis for the development of a new immune vaccine and detection reagent in the later stage. The triplex gene of AP1-AP2-BP was constructed by extended PCR technique, and the tandem gene was transformed, induced, expressed and purified, and the purified protein was made into an antigen, the rabbits were immunized with it, and prepare multi-subunit antibodies. The purified IgG was obtained from the immunized serum by affinity chromatography, and the preparation of multi-subunit antibodies. The AP1-AP2-BP three Gene tandem Recombinant Engineering strains was transformed, induced, expressed. And the purified protein obtained by Affinity Chromatography. The relative molecular weight of purified protein was 65 kDa. The content of purified protein was 3.3 mg/mL. The purified polyunit antigen had good immunogenicity. The results of EILSA showed that the titer of antibody in the serum reached 1∶5600 after the fourth immunization. The measured data of Protein A280 showed that the content of purified IgG was as high as 9.1 mg/mL. The purified protein AP1-AP2-BP multi-subunit antigen obtained from the AP1-AP2-BP three Gene tandem Recombinant Engineering strains of bovine mastitis Streptococcus agalactiae has good immunogenicity, which provides a good polyvalent antigen for the preparation of the corresponding multi-subunit antibody. At the same time, it provides a scientific basis for the development of new engineering vaccines and detection reagent.