Transgenic Cucumis sativus Expressing the Hepatitis B Surface Antigen

Abstract

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.

Virtual Screening of Indonesian Herbal Database for Cp Allosteric Modulator of Hepatitis B Virus

“…In the first position was sanggenol compound O and in the second position was cucumerin A, contained in the plant Cucumis sativum, commonly referred to as cucumbers in Indonesia. Research has shown that cucumber is an ideal plant for HBsAg expression to be used as an oral vaccine [7]. In third place was yuehchukene compound, and in the fourth place was palmarumycin CP1 compounds derived from the Jatropha curcas plant in Java (known as Jarak).…”

Production of vaccines for treatment of infectious diseases by transgenic plants

“…Many of HBsAg expression systems are still undergoing the process of development and optimisation and new plant species are being tested as possible expression systems. The most recent experiments include cucumber (Unni and Soniya, 2010), sandalwood embryogenic cell suspension cultures (Shekhawat et al, 2010), maize (Hayden et al, 2012;Hayden et al, 2015;Shah et al, 2015) and lettuce (Pniewski et al, 2011), as well as the most recent progress in using potato as host system (Rukavtsova et al, 2015). Even though the first effective parental hepatitis B vaccine by yeast has been developed more than 20 years ago, the vaccine still isn't accessible, effective or is costprohibitive for a lot of world's population, especially for areas that need it the most.…”

Expression and functional validation of heat-labile enterotoxin B (LTB) and cholera toxin B (CTB) subunits in transgenic rice (Oryza sativa)

“…Similarly, mammalian cell cultures are expensive, hindering production of large quantities of vaccines. The development of various plant expression systems has enabled the expression of foreign antigens in plant tissue as edible vaccine vehicles (Haq et al 1995 ; Du et al 2005 ; El Adab et al 2007 ; Ko et al 2009 ; Sim et al 2009 ; Unni and Soniya 2010 ; Cui et al 2011 ; Guo et al 2012 ). Plant-based edible vaccines offer advantages of longer shelf-life, stability at room temperature without loss of immunogenicity, and reduced production costs (Streatfield et al 2001 ).…”