牛分枝杆菌重组蛋白TB27.4的表达、纯化及活性鉴定

Abstract

Abstract: In order to evaluate the diagnostic properties of protein TB27.4 against bovine tuberculosis, the gene coding for TB27.4 was amplified from Mycobacterium bovis (strain Vallee Ⅲ) genomic DNA by PCR, then cloned into pET-32a(+). The recombinant plasmid was transformed into E.coli BL21(DE3) and parameters for protein expression and purification were optimized. Results of SDS-PAGE and Western blotting showed that the recombinant protein was expressed in soluble form, and could be recognized by serum from Mycobacterium bovis-infected cattle. During ELISA, the recombinant TB27.4 could specifically stimulate the PBMCs of Mycobacterium bovis-infected cattle to secrete high amount of IFN-γ. These results indicated that the recombinant protein TB27.4 had good reactogenicity and T cell activity.This study laid the foundation for further study of the role which the recombinant protein TB27.4 played in the diagnosis of bovine tuberculosis.

Key words: Mycobacterium bovis; TB27.4; expression; purification; activity identification

GAO Xin-tao, LI Ping-jun, XIN Ting, JIA Hong, GUO Xiao-yu, ZHANG Gai-mei, LI Ming, LIU Shu-qing, ZHU Hong-fei, XU Lei. Expression, Purification and Immunological Activity Identification of Recombinant Proteins TB27.4 of Mycobacterium bovis[J]. .

References

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Expression, Purification and Immunological Activity Identification of Recombinant Proteins TB27.4 of Mycobacterium bovis

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