牛分枝杆菌Mb1230基因的表达纯化及其活性评价

doi:10.16431/j.cnki.1671-7236.2015.10.005

Abstract

Abstract: To explore the value of Mb1230 protein of Mycobacterium bovis in the diagnosis of bovine tuberculosis,we obtained the Mb1230 gene by PCR and constructed recombinant plasmid pET-22b-Mb1230.Recombinant Mb1230 protein was obtained by IPTG induction and purified by affinity chromatography.The activity of the recombinant protein was evaluated by TST test,IGRA test and indirect ELISA.The size of the recombinant protein matched with the theoretical value proved by SDS-PAGE;Western blotting result showed that the recombinant protein could react with mouse anti-His antibody,and had specific band;The results of TST test,IGRA test and indirect ELISA test also showed the recombinant protein had antigenic activity.The results indicated the recombinant protein Mb1230 had good B cell and T cell activity,so,it had the potential application in the diagnosis of bovine tuberculosis.

Key words: Mycobacterium bovis; Mb1230; prokaryotic expression; diagnosis of bovine tuberculosis

CLC Number:

S852.61⁺8

LI Ming, JIA Hong, XIN Ting, GUO Xiao-yu, YUAN Wei-feng, HOU Shao-hua, HOU Qiang, GAO Xin-tao, WU Jing, DONG Rui-kai, YANG Hong-jun, LIU Lai-xing, ZHU Hong-fei. Expression,Purification and Activity Evaluation of Mb1230 Gene of Mycobacterium bovis[J]. , 2015, 42(10): 2544-2550.

References

[1] Cosivi O,Meslin F X,Daborn C J,et al.Epidemiology of Mycobacterium bovis infection in animals and humans,with particular reference to Africa[J].Rev Sci Tech,1995,14:733-746.
[2] O'Reilly L M,Daborn C J.The epidemiology of Mycobacterium bovis infection in animals and human:A review[J].Tuber Lung Dis,1995,76(Suppl 1):1-46.
[3] Smith N H,Kremer K,Inwald J,et al.Ecotypes of the Mycobacterium tuberculosis complex[J].J Theor Biol,2006,239(2):220-225.
[4] Cosivi O,Grange J M,Daborn C J,et al.Zoonotic tuberculosis due to Mycobacterium bovis in developing countries[J].Emerg Infect Dis,1998,4:59-70.
[5] Delahay R J,De Leeuw A N,Barlow A M,et al.The status of Mycobacterium bovis infection in UK wild mammals:A review[J].Vet J,2002,164(2):90-105.
[6] Morris R S,Pfeiffer D U,Jackson R.The epidemiology of Mycobacterium bovis infections[J].Vet Microbiol,1994,40:153-177.
[7] Cousins D V,Wilton S D,Francis B R,et al.Use of polymerase chain reaction for rapid diagnosis of tuberculosis[J].J Clin Microbiol,1992,30:255-258.
[8] Rothel J S,Jones S L,Corner L A,et al.A sandwich enzyme immunoassay for bovine interferon-gamma and its use for the detection or tuberculosis in cattle[J].Aust Vet J,1990,67:134-137.
[9] Bellsle J T,Vissa V D,Sievert T,et al.Role of the major antigen of Mycobacterium tuberculosis in cell biogenesis[J].Scince,1997,276(5317):1420-1422.
[10] Cole S T,Brosch R,Parkhill T,et al.Decipher the biology of Mycobacterium tuberculosis from the complete genome sequence[J].Nature,1998,393:537-544.
[11] Louise R,Skjøt V,Agger E M,et al.Antigen discovery and tuberculosis vaccine development in the post-genomic era[J].Scand J Infect Dis,2001,33(9):643-647.
[12] Alderson M R,Bement T,Day C H,et al.Expression cloning of an immunodominant family of Mycobacterium tuberculosis antigens using human CD4⁺ T cells[J].J Exp Med,2000,191(3):551-560.
[13] Renwick A R,Lightbody K L,Veverka V,et al.Structure and function of the complex formed by the tuberculosis virulence factors CFP-10 and ESAT-6[J].EMBO J,2005,24:2491-2498.
[14] Parthasarathy S,Veerasami M,Appana G,et al.Use of ESAT-6-CFP-10 fusion protein in the bovine interferon-gamma ELISPOT assay for diagnosis of Mycobacterium bovis infection in cattle[J].J Microbiol Methods,2012,90:298-304.
[15] Flores-Villalva S,Suárez-Güemes F,Espitta C,et al.Specificity of the tuberculin skin test is modified by use of a protein cocktail containing ESAT-6 and CFP-10 in cattle naturally infected with Mycobacterium bovis[J].Clin Vaccine Immunol,2012,19:797-803.
[16] Xu J,Laine O,Masciocchi M,et al.A unique Mycobacterium ESX-1 protein co-secretes with CFP-10/ESAT-6 and is necessary for inhibiting phagosome maturation[J].Mol Microbiol,2007,66:787-800.

Expression,Purification and Activity Evaluation of Mb1230 Gene of Mycobacterium bovis

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