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Development of a secretory DNA vaccine construct encoding surface layer protein EA1 of Bacillus anthracis

3rd World Congress on Biotechnology

T. G. Sumithra, V. K. Chaturvedi and P. K. Gupta

Posters: Agrotechnol

DOI: 10.4172/2155-952X.S1.020

Abstract
The quick onset, rapid lethal progression and potential to be used as biological weapon necessitate the development of a highly efficacious well tolerated vaccine against anthrax. But, currently available anthrax vaccines are so far from ideal. DNA vaccines may be attractive approaches since they are safe, stable and can induce both cellular and humoral immunity. Immune response against DNA vaccines can be increased by using secretory antigens. EA1, the major surface layer protein of Bacillus anthracis is abundant in vegetative cell surface and spore preparations. Immunity to EA1 contributes to protection at the early phase of anthrax infection making it a novel vaccine candidate. Keeping these facts, an attempt was made to develop a secretory DNA vaccine encoding EA1 of B. anthracis. For this, eag gene (2502bp) was amplified from genomic DNA of B. anthracis strain 34F2 obtained from Division of Biological Standardization, IVRI, Izatnagar using self designed forward and reverse primers having NheI and BamHI sites respectively. The purified amplicon was cloned into Signal pIg vector to create CD33 signal sequence prior to eag. The positive clones were confirmed by PCR and double RE digestion and sequence analysis. Further, cd33- eag was amplified from recombinant Signal pIg using specific primers having XbaI and NotI restriction site and was subcloned into eukaryotic expression vector, pIRES. The positive clones were confirmed by PCR and double restriction digestion. Further studies are now required to test the immunogenic potential of this DNA vaccine.
Biography
Sumithra T. G. completed her B.V.Sc & A.H in 2008 from College of Veterinary and Animal Sciences, Kerala. She received Junior Research Fellowship from Indian Council of Agricultural Research to pursue post graduation in Veterinary Bacteriology and Mycology at Indian Veterinary Research Institute, Deemed University, Izatnagar. She is presently pursuing her PhD in the same discipline as ICMR-SRF from Indian Veterinary Research Institute, Deemed University, Izatnagar.
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