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Sindhu , C. Unni and E. V., Soniya (2010) Transgenic Cucumis sativus Expressing the Hepatitis B Surface Antigen. Plant Molecular Biology Reporter, 28 (4). pp. 627-634. ISSN 1572-9818

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A DNA fragment encoding the hepatitis B virus surface antigen was amplified from a positive blood (hepatitis B) sample and introduced into the pET 32c prokaryotic expression vector. The gene encoding the HBV surface protein antigen was introduced into pCAMBIA 3300, and immobilized into Agrobacterium tumefaciens strain LBA4404. Cotyledonary leaf sections of Cucumis sativus (cucumber) cv ‘Swarnamukhi’ were cocultivated with Agrobacterium harboring the binary vector pCAMBIA 3300 carrying the HBV surface antigen gene driven by the CaMV35S promoter and the herbicide resistance gene phosphinothricin. Putative transformed shoots were induced on a Murashige and Skoog (MS) medium containing phosphinothricin, and these were then rooted on MS basal medium supplemented with 1 mg/L Indole 3-butyric acid (IBA). Integration of the T-DNA into in putative transgenic plants was confirmed by PCR and Southern blot analyses. RT-PCR and Northern blot analyses were conducted to determine RNA expression. Levels of expression in transgenic plants were confirmed by Western blot analysis, and quantification of the protein was determined by enzyme linked immuno assay (ELISA). Molecular mass of the recombinant protein was measured by Matrix-Assisted Laser Desorption Ionization-Time-of-Flight (MALDI-TOF) Mass Spectrometry.

Item Type: Article
Uncontrolled Keywords: Agrobacterium-mediated transformation Cucumis sativus Hepatitis B surface antigen Plant-based vaccine Transgenic plants
Subjects: Plant Disease Biology & Biotechnology
Depositing User: Rgcb Library
Date Deposited: 02 Sep 2019 11:27
Last Modified: 02 Sep 2019 11:27
URI: http://rgcb.sciencecentral.in/id/eprint/858

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