Chloroplast-based inducible expression of ESAT-6 antigen for development of a plant-based vaccine against tuberculosis

Publication date: Available online 24 August 2019Source: Journal of BiotechnologyAuthor(s): Kiran Saba, Johanna Gottschamel, Iqra Younus, Tahira Syed, Kehkshan Gull, Andreas Günter Lössl, Bushra Mirza, Mohammad Tahir WaheedAbstractMycobacterium tuberculosis causes tuberculosis in humans. The major disease burden of tuberculosis lies in developing countries. Lack of an effective vaccine for adults is one of the major hurdles for controlling this deadly disease. In the present study, 6 kDa early secretory antigenic target (ESAT-6) of M. tuberculosis was inducibly expressed in chloroplasts of Nicotiana tabacum. The expression of ESAT-6 in chloroplasts was controlled by T7 promoter that was activated by nuclear-generated signal peptide. Tobacco plants, containing nuclear component, were transformed via biolistic bombardment with pEXP-T7-ESAT-6 obtained by Gateway® cloning. Transformation and homoplasmic status of transplastomic plants was confirmed by polymerase chain reaction and Southern blotting. Plants were induced for protein expression by spraying with 5% ethanol for 1 day, 3 days, 7 days and 10 days. ESAT-6 protein was detected by immunoblot analysis and maximum protein was obtained for 10 days induced plants that was estimated to accumulate up to 1.2% of total soluble fraction of protein. Transplastomic plants showed completely normal morphology. Transplastomic and untransformed plants became slightly chlorotic upon prolonged exposure to ethanol until...
Source: Journal of Biotechnology - Category: Biotechnology Source Type: research

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