Using the genomic DNA of Mycobacterium tuberculosis strain H37Ra as a template,the HspX gene was amplified by PCR technology and cloned into the plasmid pET28a by seamless DNA cloning technology,and the recombinant expression plasmid pET28a-Hspx was constructed.The pET28a-HspX was transformed into Escherichia coli expression strain BL21(DE3)and HspX protein expression was induced by different temperatures,IPTG concentrations and times.The target HspX protein was purified by Ni-IDA affinity chromatography,imidazole was removed by dialysis,and the antigen specificity of HspX was detected by Western blot.Finally,the optimal induction conditions for the expression of recombinant protein HspX were determined as follows:induction temperature 37℃,IPTG concentration 0.2 mmol/L,induction time 8 h.In conclusion,this study obtained soluble HspX protein with high purity and specific recognition,laying a foundation for the future use of HspX protein in TB diagnostic reagents and vaccines.
维普
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