• The International Journal of Biological Research (TIJOBR)- Published Quarterly
  • The International Journal of Global Sciences (TIJOGS) -Published Quarterly

Isolation of SODA gene from Mycobacterium tuberculosis and its expression through Escherichia coli.

The International Journal of Biological Research (TIJOBR)


Saad Yousaf Cheema*, Mishaim Khalid, Saba Gulzar and Nimra Malik.

Institute of Molecular Biology and Biotechnology (IMBB),

 The University of Lahore, Defense Road Campus, Lahore, Pakistan.

*Corresponding Author: Email: saadyousafcheema@gmail.com 

Submitted Accepted Published
Oct 27,2019 Dec 12,2019 Jan 13,2020

2020 / Vol: 3 / Issue: 2


Mycobacterium tuberculosis is the major cause of tuberculosis in humans. It is a non-motile bacteria requiring high level of oxygen for growth. Infection starts when bacteria enters into lungs through inhalation, it grows very slowly, and the infection is mostly caused in people with compromised immune system. Superoxide dismutase (SOD) is a protein present in Mycobacterium tuberculosis. SOD is a protective protein present in bacteria, which protects bacteria from poisonous impact of reactive oxygen. SOD fixes radicals of free oxygen molecules present in bacteria. There is an iron co-factored SOD, also known as SODA, which is encoded by Rv3846 gene, and is 624bp long with molecular mass of 23 kDa. SODA is necessary for viability, and it is secreted in large amount. For cloning of SODA 624bp, reverse and forward primers were designed, which were 25bp each, then amplified with annealing temperate of 60oC. Further amplified gene was purified from gel with phenol/chloroform method and geneJet column kit method. Then E. coli were grown in LB broth and pet28avector was isolated. Then both vector and amplified PCR product were digested with Hind III and Nde I, and then ligated. The recombinant plasmid having SODA gene was successfully transformed into E. coli cells and their presence was conformed through colony PCR of transformed colonies. The work done in this project would provide easy expression of SODA protein. The SODA is potential candidate for rapid detection of tuberculosis through immunoassay. 

Keywords: Superoxide dismutase, Cloning, Mycobacterium Tuberculosis, Escherichia coli, Polymerase Chain reaction method (PCR). 


  1. Ait-Khaled, N., Enarson, D.A. (2003). Stop tuberculosis initiative. Tuberculosis: a manual for medical students. WHO.
  2. Allen, S.S., Evans, W., Carlist,e J., Hajizadeh, R., Nadaf, M., Shepherd, B.E. et al. (2008). Superoxide dismutase A antigens derived from molecular analysis of sarcoidosis granulomas elicit systemic Th-1 immune response. Respiratory Research. doi: 10.1186/1465-9921-9-36.
  3. Corbett, L. and Raviglione, M. (2005). Tuberculosis and the Tubercle Bacillus, Chapter 1 Global Burden of Tuberculosis. ISBN 1:55581(295)3.  
  4. Crick, D.C., S. Mahapatra and P.J. Brennan. (2001). Biosynthesis Of the arabinogalactan‐peptidoglycan complex of Mycobacterium tuberculosis. Glycobiology. 9: 107-118  
  5. Dye, C., Scheele, S. and Dolin, P. (1999). Consensus statement. Global Burden of tuberculosis: estimated incidence, prevalence,and mortality by country. WHO Global Surveillance and Monitoring Project. JAMA 282(7): p 677-86.
  6. Edwards, K.M., Cynamon, M.H, Voladri, R.K., Hager, C.C., DeStefano, M.S., Tham, K.T. et al. (2001). Iron-cofactored superoxide dismutase inhibits host responses to mycobacterium tuberculosis. American Journal of Respiratory and Critical Care Medicine. 15;164(12):2213-9.  
  7. Franzon, V.L, Arondel, J., Sansonetti, P.J. (1990). Contribution of superoxide dismutase and catalase activities to Shigella flexneri pathogenesis. Infection and Immunity. 58(2): 52935  
  8. Furlanetto, L., Conte-Junior, C.A., Figueiredo, E.E.S., Paschoalin, V.M.F., Silva, J., Duarte, R.S., Lilenbaum, W. et al. (2014). HPLC protocol for identification of Mycobacterium spp. From clinical sample of human and veterinary. Journal of Microbiology Research. 4. 193-200
  9. Hassett, D.J., Woodruff, W.A., Wozniak, D.J., Vasil, M.L., Cohen, M.S., Ohman, D.E. (1993). Cloning and characterization of the Pseudomonas aeruginosa SODA and SODB genes encoding Maganese and iron-Cofactored superoxide dismutase: demonstration of increased manganese superoxide dismutase activity in alginate-producing bacteria. Journal of Bacteriology. 175(23): 7658-7665 
  10. Heifets, L. and Demsond, E. (2005). Tuberculosis and the Tubercle Bacillus, Chapter 4 Clinical Mycobacteriology (Tuberculosis) Laboratory: Services and Methods. ISBN (1)55581:2953.  
  11. Khan, J., Malik, A. and Hussain, H. (2003). Tuberculosis diagnosis and treatment practices of private physicians in Karachi, Pakistan. Eastern Mediterranean Health Journal. 9(4):769–75. 
  12. Kleinnijenhuis, J., Oosting, M., Joosten, L.A., Netea, M.G., Van Crevel, R. (2011). Innate immune recognition of Mycobacterium Tuberculosis. Clinical and developmental immunology. 2011;2011: 405310. doi: 10.1155/2011/405310. Epub 2011 Apr 7.
  13. Liao, D., Fan, Q., Bao, L. (2013). The role of superoxide dismutase in the survival of mycobacterium tuberculosis in macrophages. Japanese Journal of Infectious Diseases. 66(6): 480-8
  14. Pierce Marcia, M. (2010). New drug to control tuberculosis. AccessScience McGrew hill education. 
  15. Stanley, R.L., Sterling, T.R., Thompson, D., McElroy, P.D., Madison, A., Moore, K. et al. (2000). A multi-state outbreak of tuberculosis among members of a highly mobile social network: implications for tuberculosis elimination. International Journal of Tuberculosis and lung disease 4(11): 1066-73   
  16. Todar, K. (2011). Todar’s online textbook of bacteriology. Chapter: bacterial pathogens and diseases of humans; tuberculoses
  17. World health organization (WHO). Global Tuberculosis report (2016).   www.who.int/tb/publications/global_report/en/  date last accessed 1st August 2017.  
  18. Zhang , Y., Lathigra, R., Garbe, T., Catty, D., Young, D. (1991). Genetic analysis of superoxide dismutase, the 23 kilodalton antigen of mycobacterium tuberculosis. Molecular Microbiology. 5(2): 381-91.

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