Mutation Analysis And Quantitative Proteomics Analysis Of Mycobacterium Bovis Virulence Gene

Tuberculosis(TB)is a chronic wasting infectious disease that kills two million people worldwide each year.The main pathogens of tuberculosis are Mycobacterium tuberculosis and Mycobacterium bovis.Bovine tuberculosis is a major problem affecting the economy and animal health,as well as a threat to human health.The prevalence of bovine TB is estimated at 9%worldwide,particularly in developing countries.Bovine tuberculosis has a direct impact on animal production performance and the economic benefits of cattle ranching.However,antibiotics are widely used in humans and animals,leading to the emergence of multi-drug resistant strains,causing the bacteria to change dramatically when they adapt to changes in the environment and have a huge impact on human health.Therefore,in this study,gene sequencing and proteomics were used to investigate and analyze the pathogenicity and virulence of different M.bovis from the gene level and protein expression level.The specific test methods and results are as follows:1.In order to study the mutation level of the M.bovis virulence gene,analysis of 10 virulence genes of two strains of M.bovis and 66 clinical isolates with different pathogenicity.The 10 virulence genes after screening were subjected to PCR identification,sequencing Blast,SNPs evaluation and protein secondary structure function prediction.The results showed that the 66 isolates had 10 virulence genes that were highly consistent with the N and had a high genetic similarity,reflecting the strong adaptability of the N in cattle.Through software analysis and prediction,it was proved that changes in the secondary structure of the protein caused by genetic variation of the gene may change the function of the protein.In the end,the genes MB2982c,ctpc,espi may have a greater impact on the virulence of bacteria,and further research is needed.2.In order to study the differential expression of protein levels between N and C68004,quantitative proteomics approach was used to identify.The results of GO analysis showed that DEPs included catalysis,binding,transport,transcriptional regulation,and molecular structure-related effects,especially in response to internal or external stimuli.The results of KEGG analysis showed that EDPs were mainly involved in the regulation of metabolism and biological processes.The analysis screened out the virulence genes that may cause the pathogenicity of N were mmaA4,eccel,IpqY,hspX,Mpb63,mmpl4.The expression levels of resistance genes rpoA,rpoB and rpoC of N were significantly higher than that of C68004,which can be inferred that the resistance of N bacteria against the first-line TB drugs rifampicin and isoniazid may be stronger than C68004.In summary,the pathogenicity of Mycobacterium bovis may be related to mutations in virulence genes.At the same time,N has a strong adaptability in cattle,which can represent the prevalence of M.bovis and the evolutionary trend of strains in some regions.With the frequent use of clinical drugs and changes in immune response,virulence genes have changed during the adaptation of the N,and protein expression related to metabolism,DNA replication,and drug resistance have also appeared different.