Study On The Separation,Purification And Activity Analysis Of MT3286 Protein From Mycobacterium Tuberculosis

Tuberc?losis is a chronic consumption which has strong injuries on h u man and animals' healthy.Mycobacterium tuberculosis and mycobacteri?m bovis are the main pathogenic bacteria that can cause tuberculosis.The widespread of immunodeficiency with AIDS,the consequence of poverty on sanitary protection and information at both individual and state levels lead control of tuberc?losis(TB)to be one of the priorities of World Health Organization programs.With the emergence of Drug-resistant strains of Mycobacterium tuberculosis,the vast majority of first-line anti-TB drugs have emerged in varying degrees of resistance,which result the tuberculosis has a rebound in the global trend.In the 1990's,tuberc?losis epidemic disease had risen significantly in 41.5%developing countries and 25%development countries.PDZ domain is one of the common domains which are involved in diverse protein-protein interactions and many biological processes(such as cell signaling,cell polarity,cell migration,invasion and cell recognition).Therefore,the studies on PDZ domain protein are conducive to realize the invasion and diffuse process of Mycobacterium tuberculosis in vivo.Thus,we can develop a new generation of tuberc?losis targeted drugs and vaccines.In this study,the gene of MT3286 comes from Mycobacterium tuberculosis CDC1551,which is homologous in H37Rv and BCG genome 100%and there has been no word on the structure and function of MT3286.Characteristics of the gene and vector,the gene was cloned into pET28a vector,and then transformed into Esherichia coli BL21 to express fusion protein.After a series of condition optimization,we had got the high-purity protein of MT3286 and the molecular weight coincides with the predetermination.Followed by the using the Ni-NTA affinity chromatography and Superdex 200 gel chromatography methods for the protein's separation and purification.The interaction with type-I collagen,hyaluronic acid,chondroitin s?lfate,calf thymus DNA,maltose,sorbitol,farina and BSA were measured.The res?lts show that MT3286 and type-I collagen,hyaluronic acid,chondroitin sulfate generate floccose sediment respectively when incubation more than three hours at 39?.The inspection by SDS-PAGE found that the constituents in sediment were compound of MT3286 and type I collagen.A protein of Mycobacterium tuberculosis was expressed successfully and adhesions(Ingredients of Mycobacterium tuberculosis,which participate in the combination with host cells and extracell?lar matrix)of Mycobacterium tuberculosis activities were detected in this study.The results of this research provide basis for in-depth structure and functional biology study.