Preparation And Application Of Monoclonal Antibodies Of Early Secreted Proteins MPB 70 And MPF 83 Of Bovine Mycobacterium Tuberculosis

Mycobacterium,which has been confirmed by research to be a type of Gram-positive acid-fast bacilli,lives in the environment of daily life as a bacterium.Mycobacteria can also be divided into many types,one of which can form a Mycobacterium tuberculosis complex because of common antigen proteins.Most strains grow slowly,and some can cause tuberculosis and nucleoid diseases in humans and animals.The specific secreted proteins of early infection that are unique to Mycobacterium bovis tuberculosis are MPB 70 and MPB 83 proteins,which are mainly used in the identification of strains and vaccine research.Based on the comprehensive analysis of the early identified secreted proteins MPB 70 and MPB 83 of M.bovis tuberculosis using DNAstar7.0 software,the major antigenic domains of the MPB 70 and MPB 83 protein coding genes were screened,using overlapping extension PCR technology,MPB 70 and MPB 83 tandem recombinants were constructed.It was ligated into the prokaryotic expression vector pGEX-6p-1 for induction and expression.An antigenic tandem antigen of MPB 70,MPB 83 secreted proteins of Mycobacterium bovis infection was obtained.Using DNAStar bioinformatics software to predict the structural regions of MPB 70 and MPB 83 antigens,which are early markers of Mycobacterium tuberculosis infection.And designed and synthesized cloning primers,constructed the recombinant vector and obtained high expression.Coupled with the smooth preparation of monoclonal antibodies,the purity of the purified recombinant protein and antibody titer expression were detected.In this experiment,the secreted protein tandem antigen MPB 70+MPB 83 of Mycobacterium bovis infection with obvious antigenic activity was obtained,which laid a solid foundation for the further development of a rapid detection kit for early infection markers of Mycobacterium bovis tuberculosis.