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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
Index Copernicus ICI Journals Master List 2018 - IJCMAS--ICV 2018: 95.39 For more details click here
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2018: 95.39
For more details click here

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Original Research Articles

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2017.6(10): 1085-1093
DOI: https://doi.org/10.20546/ijcmas.2017.610.132


Expression of Brucella Lumazine Synthase Gene in E. coli System
M. Nagalingam1*, Thaslim J. Basheer1, N. Vijaya Gowri1, V. Balamurugan1, Rajeswari Shome1, G.B. Manjunatha Reddy1, B.R. Shome1, H. Rahman2, Parimal Roy1 and R.K. Gandham3
1ICAR-National Institute for Veterinary Epidemiology and Disease Informatics,Bengaluru, Karnataka 560 064, India
2International Livestock Research Institute, New Delhi 110 012, India
3ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243122, India
*Corresponding author
Abstract:

Brucellosis is an important zoonosis of serious economic and public health consequences affecting livestock and human beings caused by Brucella spp. The available whole cell antigen and smooth lipopolysaccharide (sLPS) based diagnostics show false positive reactions due to cross reactivity. Therefore, in this study, Brucella lumazine synthase (BLS), an immunogenic protein of Brucella spp. coding 477 bp bls gene was amplified from DNA extracted from Brucella abortus S99 strain and the amplified product was cloned in pET directional TOPO expression vector and transformed in E. coli TOP 10F’ cells. After verifying sequence, bls recombinant clones were subsequently transformed in E. coli BL21 (DE3)/pLysS cells for expression. BLS protein expression was induced with 1mM IPTG and optimized, six hours induction yielded maximum expression of BLS protein. The expressed protein was purified by His-tag affinity purification method using Ni-NTA and characterized by SDS-PAGE and Western blot. Recombinant BLS protein expressed in E. coli has potential use as diagnostic antigen or immunogen for development of diagnostics and vaccine for brucellosis.


Keywords: Brucellosis, Brucella Lumazine Synthase (BLS), Recombinant protein, TOPO vector.
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How to cite this article:

Nagalingam M., Thaslim J. Basheer, N. Vijaya Gowri, V. Balamurugan, Rajeswari Shome, G. B. Manjunatha Reddy, B. R. Shome, H. Rahman, Parimal Roy and Gandham R. K. 2017. Expression of Brucella Lumazine Synthase Gene in E. coli System.Int.J.Curr.Microbiol.App.Sci. 6(10): 1085-1093. doi: https://doi.org/10.20546/ijcmas.2017.610.132